Immunization with CoronaVac or ChadOx-1 Elicit Distinct Modulation of Monocyte Subtypes and Systemic Inflammatory Status in Older Adults

Immunization with CoronaVac or ChadOx-1 Elicit Distinct Modulation of Monocyte Subtypes and Systemic Inflammatory Status in Older Adults | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Immunization with CoronaVac or ChadOx-1 Elicit Distinct Modulation of Monocyte Subtypes and Systemic Inflammatory Status in Older Adults Fernanda Rodrigues Monteiro, Ariane Nardy, Vitória Paixão, Jônatas Bussador Amaral, and 11 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7085263/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background The older adult population was significantly affected by COVID-19 due to immunosenescence, which alters immune elements such as monocytes, impacting immune responses to pathogens and vaccination. Aging-associated systemic inflammation, or inflammaging, can further impair vaccine responses. Although important data exist, the effects of COVID-19 vaccination on monocytes and systemic inflammation in older adults are not fully understood. This study aimed to investigate the impact of COVID-19 vaccination with CoronaVac and ChAdOx-1 on monocyte subtypes and systemic inflammatory status in older adults. Methods Sixty-six older adults (74.74 ± 5.9 years) participated in this study. Blood samples were collected before and 30 days after the second dose of either CoronaVac (n = 48) or ChAdOx-1 (n = 18) to measure IgG levels for SARS-CoV-2, cytokines, and monocyte subtypes (classical, non-classical, and intermediate). Results Both vaccines increased specific IgG levels post-vaccination. The ChAdOx-1 group exhibited higher IgG levels, more classical, non-classical, and intermediate monocytes, and higher IL-6/IL-10 and TNF-α/IL-10 ratios compared to CoronaVac. Conversely, CoronaVac led to higher IFN-γ post-vaccination. Both vaccines promoted distinct modulation of monocyte subtypes and inflammatory status. Conclusion Both vaccines elicited a strong antibody response, but ChAdOx-1 demonstrated better results. Distinct changes in monocyte subtypes and inflammation were observed between the two vaccine groups. ChAdOx-1 CoronaVac cytokines IgG inflammation SARS-CoV-2 Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Background Among some characteristics, the older adult population is one of the most affected groups by coronavirus disease 19 (COVID-19) [ 1 , 2 ], and the increased susceptibility and severity in this population are attributed to immunosenescence [ 3 , 4 ], a phenomenon associated with a gradual decline in immune cell functions, including monocytes [ 5 ]. Monocytes are crucial in the inflammatory process and are classified into three subtypes: classical (CD14 + + CD16-), non-classical (CD14 + CD16++), and intermediate (CD14 + + CD16+) [ 6 – 15 ]. Classical monocytes, comprising around 90% of total circulating monocytes, are considered inflammatory cells and produce several inflammatory cytokines [ 6 – 9 ]. Non-classical monocytes, constituting approximately 10% of circulating monocytes, exhibit an anti-inflammatory phenotype and are more responsive to virus-associated stimuli [ 10 , 13 ]. Intermediate monocytes, representing up to 5% of circulating monocytes, display high phagocytic capacity and antigen presentation ability [ 14 , 15 ]. Aging is associated with a tendency towards higher proportions of non-classical monocytes, although it was shown that monocytes from older adults exhibit increased basal cytokine production compared to those from younger adults. Additionally, non-classical monocytes display characteristics of cellular senescence, suggesting a link between aging and changes in monocyte subset proportions [ 16 ]. It is important to mention that the raised expression of genes related to inflammation by monocytes of aged people drives the decreased vaccination responses, indicating a marked interplay between the elevation of the monocyte number and the inflammatory status, which can negatively impact vaccination responses [ 17 , 18 ]. Regarding the COVID-19 vaccination in Brazil, it was available four types of vaccines with different mechanisms of action, such as the mRNA vaccine from Pfizer/BioNTech (BNT16b2) and Moderna (mRNA-1273), or non-recombinant viral vector vaccines from AstraZeneca/Oxford (ChAdOx-1, AZD1222) and Janssen-Cilag (Ad26.COV2.S), or whole-inactivated virus vaccine from Sinovac/Butantã (CoronaVac). It is worth citing that among these, only CoronaVac does not specifically use the viral envelope glycoprotein called spike as the specific target [ 19 ]. Although monocytes play a pivotal role in increasing cytokine production and severity of COVID-19 in older adults, as well as in vaccination responses [ 16 – 22 ], the effects of vaccination with ChAdOx-1 and CoronaVac on monocyte subtypes in older adults remain unclear. Therefore, the present study aimed to assess these effects on monocyte subtypes and circulating cytokine levels. Methods Population and Design of the Study In this observational study, at two phases (pre- and post-vaccination), 66 older adults, aged between 60 and 85 years, of both sexes, and submitted to vaccination for SARS-CoV-2 were enrolled. All participants were recruited from the Older Adults Health Promotion Outpatient Clinic belonging to the Discipline of Geriatrics and Gerontology at the Federal University of São Paulo (UNIFESP), and follow-up for the same geriatrician. As shown in Fig. 1, clinical and physical data, and blood samples were obtained between January and February 2021 (before the first round of vaccination) and between March and April 2021, which corresponds to 30 days after the second dose of the CoronaVac vaccine, or between July and August 2021, which corresponds to 30 days after the second dose of the ChAdOx-1 vaccine. The present study followed all the guidelines of the Declaration of Helsinki [ 23 ]. Volunteers did not exhibit respiratory or any other symptoms during the study and received two COVID-19 vaccine doses (inclusion criteria). Participants who presented positive serology for HIV, frailty, sarcopenia, cancer, thrombosis, renal, liver, and neurological diseases, or reported use of corticosteroid and anti-inflammatory medication, or convalescent plasma therapy before and during the study, or even did not attendance at sample collection sessions were excluded. Sample Size Calculation Groups Using G*Power software, sample size and statistical power were estimated for a Student's T-test with an effect size of 0.30, α level of 5%, and a statistical power of 0.95 for two groups with pre- and post-vaccination measurements. Based on previous studies [ 24 , 25 ], a minimum of 55 individuals was considered after accounting for a 30% margin of losses or refusals. Initially, 92 older adults were invited and agreed to participate, but by inclusion/exclusion criteria, 81 volunteers were eligible. However, 15 volunteers were excluded because they did not attend sample collection. Therefore, data and samples from two groups: one immunized with CoronaVac (n = 48) and the other with ChAdOx-1 (n = 18) were used here. Vaccination for COVID-19 In according to the vaccination schedule initially proposed for the aged population by the National Health Surveillance Agency (ANVISA) of Brazil, in January and February 2021, particularly this population could be immunized exclusively with the vaccine CoronaVac, which was produced with the whole inactivated virus. After two weeks, it was also available the ChAdOx-1/AstraZeneca (AZD1222), which was a chimpanzee non-replicating adenovirus vector vaccine encoding the spike glycoprotein of SARS-CoV-2 into the host cells. All volunteers received two doses of the same vaccine, but the interval between the first and second dose for the vaccination with CoronaVac was around 30 days, whereas for the ChAdOx-1 was around 120 days. Collection of Blood Samples Peripheral blood samples were collected before and 30 days after the second dose of COVID-19 vaccines (CoronaVac or ChAdOx-1) to obtain plasma and peripheral blood mononuclear cells (PBMC). Plasma aliquots were frozen at -80ºC for subsequent analysis of systemic immunoglobulin G (IgG) and pro- and anti-inflammatory cytokine concentrations. PBMC was isolated, storage and used as follows described. Determination of specific immunoglobulin G (IgG) for SARS-CoV-2 The systemic specific IgG levels for SARS-CoV-2 antigens were determined using the ELISA test in plasma samples previously storage, following the protocol described by our group [ 26 , 27 ]. Immunophenotyping of monocyte subtypes Blood samples were mixed with PBS and layered on tubes containing Histopaque. followed by centrifugation to obtain the PBMC. After, the PBMC was isolated, washed, mixed with a frozen medium, and stored in liquid nitrogen until their analysis. PBMC cells were transferred to a Falcon® tube containing 10% PBS-SFB solution, followed by centrifugation and resuspension in 1% PBS-BSA. The cells were then incubated with monoclonal antibodies conjugated to fluorochromes (CD14-FITC and CD16-PE), along with isotype controls from BD Biosciences. After incubation, the cells were washed and resuspended in PBS, and monocyte subtypes were assessed through the flow cytometer FACSCalibur (Becton Dickinson Immunodiagnostic Systems, San José, CA, USA), acquiring a minimum of 10,000 events. Data analysis was performed using BD CellQuest Pro software (Becton Dickinson Immunodiagnostic Systems, San Jose, CA, USA). Figure 2 shows the study's strategy used to define monocyte populations and non-classical (CD14 + CD16++), intermediate (CD14 + + CD16+), and classical (CD14 + + CD16-) subtypes. Determination of Cytokines Plasma concentrations of IL-4, IL-6, IL-10, IL-12p70, IL-13, TNF-α, IFN-α, IFN-β, and IFN-γ were determined using the LEGENDplex™ bead-based multiplex assay (Biolegend, San Diego, CA, USA), following the manufacturer's instructions, with a linear range of 2.4 to 10,000 pg/mL and high correlation coefficients for standard curves. Statistical analysis Firstly, both normality and homogeneity of each data were determined by the Shapiro-Wilks and Levene tests, respectively. Parametric data (demographic and anthropometrics) were represented as mean and standard deviation (X_±_SD) and were assessed by Student t-test, whereas non-parametric data (immune/inflammatory parameters) were represented as median and interquartile ranges, and the Wilcoxon test was used for intragroup comparisons, whilst the Kruskal-Wallis test with Dunn's post-hoc test was used for intergroup evaluations. Besides, Spearman´s rank correlation coefficient test was also performed. For all analyses, it was used the GraphPadPrism8 software (GraphPad Software, La Jolla, CA, USA) and set the α risk at 5% (p < 0.05). Results Table 1 presents demographic and anthropometric data of volunteer groups before and after separation in agreement with the type of vaccine received (CoronaVac or ChAdOx-1). No differences were observed in these parameters. Table 1 Anthropometric data presented as a mean and standard deviation (X ± SD) of the volunteers participating in the current study. Volunteers´ Group Variables CoronaVac (n = 48) ChAdOx-1 (n = 18) p-value Age (years) 74.0 ± 4.3 76.6 ± 8.8 0.0966 Height (meters) 1.58 ± 0.10 1.59 ± 0.11 0.7490 Weight (kg) 63.75 ± 12.36 70.15 ± 16.66 0.0941 BMI (kg/m²) 25.56 ± 4.28 27.72 ± 5.15 0.0894 Figure 3 illustrates the specific antibody (IgG) response to SARS-CoV-2 antigens, showing higher IgG levels post-vaccination than pre-vaccination values for both groups (p < 0.0001 for CoronaVac and ChAdOx-1). However, the ChAdOx-1 group exhibited higher IgG levels than the CoronaVac group (p = 0.0001). Additionally, the immunogenicity found in the ChAdOx-1 group was 72.2% (13/18), whereas in the CoronaVac group was 39.6% (19/48). It is important to mention that the immunogenicity value was set considering an increase of, at least, two-fold in IgG levels at the post-vaccination moment in comparison to the pre-vaccination values. Figure 4 presents the results, expressed in percentages, of monocytes subtypes. ChAdOx-1 group exhibited significantly increased percentages of all subtypes post-vaccination as compared to both pre-vaccination values (p < 0.0001) and the CoronaVac group (p < 0.0001). No significant differences were observed in the CoronaVac group. Figure 5 displays the data concerning the systemic cytokine levels assessed here [IL-4 (A), IL-6 (B), IL-13 (C), IL-10 (D), IL-12p70 (E), TNF-α (F), IFN-α (G), IFN-β (H), and IFN-γ (I)]. It was observed that the ChAdOx-1 group, at post-vaccination, showed lower levels of IL-4 (A), IL-13 (C), IL-10 (D), IL-12p70 (E), IFN-β (H), and IFN-γ (I) than pre-vaccination and CoronaVac group. Besides, post-vaccination IFN-γ levels (I) in the CoronaVac group were higher than pre-vaccination and the ChAdOx-1 group post-vaccination. No differences were observed in IL-6 and TNF-α. Regarding the analysis of the ratio between pro-inflammatory cytokines and the anti-inflammatory IL-10 cytokine (Fig. 6), at post-vaccination, the ChAdOx-1 group demonstrated higher IL-6/IL-10 (B) and TNF-α/IL-10 (E) ratios, along with lower IL-12p70/IL-10 (C), IL-13/IL-10 (D), and IFN-β/IL-10 (G) ratios than pre-vaccination and CoronaVac group. Additionally, the ChAdOx-1 group exhibited a higher IFN-α/IL-10 (F) ratio and lower IFN-γ/IL-10 (H) ratio post-vaccination than the CoronaVac group. No difference was observed in the IL-4/IL-10 ratio (A). Correlation analyses, detailed in Table 2 , revealed significant positive correlations between various monocyte subtypes at pre- and post-vaccination occasions in both volunteer groups. Notably, the ChAdOx-1 group exhibited specific positive correlations, including intermediate and classic monocytes with the IL-6/IL-10 ratio pre-vaccination, and non-classical monocytes with this ratio post-vaccination. Conversely, the CoronaVac group showed significant positive correlations between all monocyte subtypes and the IL-13/IL-10 ratio post-vaccination. Additionally, both groups displayed significant positive associations between systemic IgG levels and all monocyte subtypes post-vaccination. Particularly at the pre-vaccination occasion, significant correlations were found between classical monocytes and IL-12p70, as well as between intermediate and classical monocytes with IFN-α, contrasting with significant negative correlations involving intermediate and classical monocytes with IL-13, IL-10, or IFN-γ. Table 2. Spearman's correlation coefficient analysis between monocyte subtypes, total serum levels of specific IgG for SARS-CoV-2, and pro- and anti-inflammatory cytokines in volunteer groups vaccinated with CoronaVac or ChAdOx-1 before the first dose (pre) and 30 days after the administration of the second dose (post) of these vaccines. Parameters PRE Pre-vaccination Parameters CoronaVac Post-vaccination Parameters ChAdOx-1 Post-vaccination rho-velue p-value rho-value p-value rho-value p-value Intermediate x IL-13 -0.2781 0.0237 IgG x Non-classical 0.3781 0.0080 IgG x Non-classical 0.8493 <0.0001 Intermediate x IL-10 -0.3220 0.0083 IgG x Intermediate 0.3557 0.0130 IgG x Intermediate 0.8493 <0.0001 Intermediate x IFN- a 0.2680 0.0295 IgG x Classical 0.3517 0.0142 IgG x Clássical 0.8493 <0.0001 Intermediate x IFN- g -0.2840 0.0208 Non-classical x IL-13/IL-10 0.3277 0.0230 Non-classical x IL-6/IL-10 0.5550 0.0168 Intermediate x IL-6/IL-10 0.5139 <0.0001 Intermediate x IL-13/IL-10 0.3277 0.0230 Intermediate x IL-6/IL-10 0.5550 0.0168 Intermediate x TNF- a /IL-10 0.3958 0.0010 Classical x IL-13/IL-10 0.3277 0.0230 Classical x IL-6/IL-10 0.5550 0.0168 Intermediate x IFN- a /IL-10 0.3774 0.0017 Classical x IL-13 -0.3026 0.0135 Classical x IL-10 -0.3437 0.0047 Classical x IL-12p70 -0.2489 0.0438 Classical x IFN- a 0.2750 0.0253 Classical x IFN- g -0.2896 0.0183 Classical x IL-6/IL-10 0.5253 <0.0001 Classical x TNF- a /IL-10 0.4161 0.0005 Classical x IFN- a /IL-10 0.3877 0.0012 Discussion In general, our results demonstrated that immunization with the ChAdOx-1 and CoronaVac vaccines elicited significant differences in the immune/inflammatory responses in older adults. In fact, it was possible to evidence that the group vaccinated with ChAdOx-1 showed a considerable reduction in the levels of some cytokines, both pro- and anti-inflammatory properties, as well as an increase in the percentages of all monocyte subtypes as compared to the results found in the group vaccinated with CoronaVac. In addition, it was verified that volunteers in the ChAdOx-1 group presented higher specific levels of IgG to SARS-CoV-2 antigens than the CoronaVac group. This observation of higher IgG levels in the ChAdOx-1 group than in the CoronaVac group post-vaccination corroborates a previous study in which > 60-year-old individuals vaccinated with CoronaVac presented lower antibody responses than younger individuals immunized with the same vaccine and individuals with the same age immunized with a vaccine containing mRNA for the S (spike) protein of SARS-CoV-2 [ 28 ]. Moreover, it was reported that vaccination efficacy 14 days or more after administration of the second dose decreased with increasing age, for example, dropping from 61.8% among individuals aged 70 to 74 years to 48.9% among individuals aged 75 to 79 years, and to 28.0% among individuals aged ≥ 80 years [ 29 ]. Likewise, our group also formerly showed that older adults immunized with two doses of the ChAdOx-1 vaccine presented a better antibody response than the group immunized with two doses of CoronaVac vaccine and also that lower IgG levels post-vaccination were associated with age in the CoronaVac group [ 27 ]. Additionally, the observation that the ChAdOx-1 group presented higher immunogenicity than the CoronaVac group agrees with the literature [ 30 ]. Beyond these findings, the positive correlations between IgG levels and the monocyte subtype percentages in both volunteer groups can reinforce the role of monocytes in vaccine responses, including COVID-19 [ 31 ]. Although it was reported a reduced non-classical monocyte frequency in COVID-19, after the dose of vaccination, these monocytes exhibited a dual profile, concurrently expressing both pro- and anti-inflammatory cytokines, suggesting that these monocytes can act in protection against viral infection [ 32 ]. Since some vaccines can trigger a memory-like response in monocytes, it is plausible to suggest that COVID-19 vaccination could induce the training of monocytes [ 33 ]. Particularly, people immunized with the ChAdOx-1 vaccine showed that the monocyte activation was related to not only neutralization antibody levels but also increased capacity for antigen presentation and a cytokine profile that could favor the development of adaptive immunity [ 34 , 35 ]. Concerning the CoronaVac vaccine, both a significant increase in classical and intermediate monocyte percentages and a decrease in non-classical monocyte frequency were verified post-vaccination with two doses, which, according to the authors, can supposedly indicate that an antiviral and adaptive response was elicited [ 36 ]. These pieces of information can corroborate our findings that an enhancement in the percentage of all monocyte subtypes post-vaccination was positively associated with systemic IgG levels in both groups of volunteers. However, here, only the ChAdOx-1 group showed a significant increase in the monocyte subtype percentages post-vaccination. Even if we cannot affirm, this finding could be related to the technology/design used in the ChAdOx-1 vaccine, since it was a chimpanzee non-replicating adenovirus vector vaccine encoding the spike glycoprotein of SARS-CoV-2 into the host cells, thus stimulating the innate immune response to especially prime the CD4 + T cells to differentiate into effector and memory cells that will help B cells to differentiate into antibody-secreting plasma cells. Differently, the CoronaVac vaccine was designed to contain the inactivated whole SARS-CoV-2 virus, which is not only a widely used technique but can also elicit protective immunity [ 37 , 38 ]. Based on these pieces of information, seemingly, the vaccine that uses an "intracellular delivery" design, such as ChadOx-1, can be more effective in eliciting the enhancement of the percentage of all monocyte subtypes than the CoronaVac in older adults. Particularly the increased percentage of monocytes observed in older adults, in conjunction with the alteration of monocyte subsets towards intermediate and non-classical phenotypes verified during aging, could potentially correlate with elevated production of some cytokines, such as IL-6, IL-10, and TNF-α [ 39 ]. These data corroborated the positive correlations found not only between all monocyte subtype percentages at pre-vaccination, but also between classical monocytes with circulating levels of IL-13, TNF-α, and IFN-α, as well as with the ratios IL-6/IL-10 and TNF-α/IL-10, in contrast to the negative correlations with the IL-10 and IFN-γ levels. Likewise, intermediate monocytes were positively correlated with IFN-a levels and with the ratios IL-6/IL-10, TNF-α/IL-10, and IFN-α/IL-10, and negatively correlated with IL-10, IL-13, and IFN-γ levels. Therefore, these findings allow us to suggest that systemic inflammatory status can modulate the monocyte subtype percentages in older adults. Interestingly, post-vaccination, whereas the monocyte subtypes in the CoronaVac group were positively correlated with the IL-13/IL-10 ratio, in the ChAdOx-1 group this positive correlation was with the IL-6/IL-10 ratio. Concerning these findings, it is worth mentioning that it is well-known that IL-6 and IL-13, two cytokines related to the T help (Th)-2 profile, can promote B cell differentiation and subsequent antibody production [ 40 ]. Regarding the triad - cytokines/aging/vaccination - as previously mentioned the development and progression of immunosenescence and inflammaging can impair the vaccination responses [ 41 ] since an aberrant cytokine production can reduce the ability to process and present antigens to T cells, which negatively impacts the B cells' activation and antibody production [ 34 ]. Therefore, circulating cytokines profile assessment is pivotal to better characterize the immune/inflammatory responses elicited by vaccination, particularly in older adults. Despite the isolated analysis of systemic cytokine levels providing important data, it is paramount to point out that the ratio analysis can offer an accurate measure concerning the balance between pro- and anti-inflammatory cytokines in several contexts, including in aging [ 42 ]. Similarly to those findings for monocytes, a distinct and interesting signature regarding the systemic cytokine profile was evidenced in the volunteer groups post-vaccination for COVID-19. In this sense, the isolated cytokine evaluations showed, in a general way, two aspects that aroused interest: (1) reduced levels of most cytokines assessed here in the ChAdOx-1 group, whereas (2) only IFN-γ levels were reduced in the CoronaVac group. This last finding corroborates previous studies in which the number of IFN-γ-producing cells, particularly T and NK cells, in response to the recombinant S protein from SARS-CoV-2, increased around 28 days after the administration of two doses of the CoronaVac vaccine [ 43 – 45 ]. Therefore, we can suggest that this vaccine was able to elicit IFN-γ production by the immune cells, which also impacted the higher IFN-γ/IL-10 ratio found in this volunteer group. Although IFN-γ is closely related to cellular immune responses, it has been cited that this cytokine can also enhance the humoral immune responses post-immunization, especially by improving both the capacity of immune cells to perform virus clearance and vaccination efficacy [ 46 – 48 ]. Concerning the decrease in some cytokines found in the ChAdOx-1 group, this was an unexpected finding since had been reported an increase in cytokine levels post-vaccination, including in older adult individuals [ 49 , 34 ]. Despite we cannot explain why the cytokine levels were lower post-ChAdOx-1 vaccination when assessed individually, the data obtained in the ratio analysis can help us understand these findings. In this respect, the significant reductions of IL-12p70, IL-13, and IFN-β levels could be responsible for the lower ratio between these cytokines with IL-10, whereas the maintenance of IL-6, TNF-α, and IFN-α levels can drive the higher ratio between them with IL-10 found in this group. Regarding IL-6, it is a pro-inflammatory cytokine with pleiotropic actions in different contexts [ 50 , 51 ], including COVID-19 and vaccination [ 52 ]. It is worth highlighting that the maintenance of IL-6 levels in the volunteer groups was a favorable result since higher IL-6 levels were associated with both an increased risk of death [ 53 ] and the development of cytokine storm in COVID-19 patients, as well as the increase of inflammaging, which, consequently, impairs the vaccination responses [ 54 ]. Particularly in the context of aging, it was reported that IL-6 not only was involved in restoring the generation, memory, and responses of CD4 + T cells, as well as the CD8 + T cells' response to influenza virus infection from aged mice [ 55 , 56 ], but also this cytokine has a pivotal as a growth factor and for the differentiation of B cells into antibody-producing cells. 52 Additionally, a limited IL-6 production was associated with a better immune response to influenza virus vaccination [ 51 ]. Taken together, these data allow us to suggest that the "controlled" increase of the IL-6/IL-10 ratio could favor the immune response to the ChAdOx-1 vaccine in older adults. Beyond this finding, the higher ratios of TNF-α/IL-10 and IFN-α/IL-10 post-vaccination in the ChAdOx-1 group can demonstrate that a Th-1 immune profile was elicited. Concerning TNF-a, a classical pro-inflammatory cytokine, it is involved in cell-mediated immunity and phagocyte-dependent inflammation, particularly in virus infection [ 57 ], and also when produced by CD8 + T cells and NK cells (typical Th-1 cells), this molecule can help in controlling viral replication [ 58 ]. Similarly to IL-6, TNF-α overproduction has been associated with worse outcomes in COVID-19 and the development of inflammaging and immunosenescence, which includes negative effects in B cell responses [ 59 ]. Therefore, the maintenance of TNF-α levels post-vaccination is a prominent finding since can demonstrate that the inflammaging was not increased by the vaccination, and also the increased TNF-α/IL-10 ratio found in the ChAdOx-1 group allows us to suggest that a Th-1 immune response was preferentially towards, which could improve the virus clearance and, consequently, reduces the COVID-19 severity [ 60 ]. Corroborating this suggestion, it was also observed a higher IFN-α/IL-10 ratio in the ChadOx-1 group. It is paramount to mention that the different types of interferon [Type-I (IFN-α and IFN-β); type II (IFN-γ); or type-III (IFN-λ)] are exclusively related to Th-1 immune responses and have a central role in viral infections not only by promoting the control of these pathogens but also by eliciting a robust adaptive immune response that drives to an efficient viral clearance [ 61 ]. Regarding coronavirus infection, patients with SARS-CoV [ 62 , 63 ] and MERS [ 64 ] presented increased IFN-α levels, and it was suggested that this finding could be related to the improvement of not only the antigen presentation but also a robust antiviral response for these viral infections [ 65 , 66 ] since this cytokine can promote the monocyte differentiation to dendritic cell, which are pivotal for the antigen presentation to CD4 + T cells [ 26 ]. Lastly, it is worth highlighting that some limitations of this study were: (1) the discrepant number of volunteers in the CoronaVac (n = 48) and ChadOx-1 (n = 18) groups, however, as formerly cited, this difference was attributed to the fact that, although these were the first vaccines for COVID-19 available in Brazil, especially for the older adults population the CoronaVac was the main vaccine offered at the beginning of COVID-19 vaccination; (2) the lack of data concerning the activation levels of monocyte subtypes; (3) the lack of comparison of the immunophenotyping of T and NK lymphocytes, which could improve the findings of the present study, especially concerning systemic inflammatory status. Conclusion Taking together, our findings showed that older adults immunized with ChAdOx-1 presented higher specific IgG levels for SARS-CoV-2 antigens than the older adult group immunized with CoronaVac, and this difference can putatively be associated with the distinct responses observed in the percentage of monocyte subtypes and systemic inflammatory status found in these volunteer groups. Declarations Ethics approval and consent to participate Informed consent was obtained from all subjects and/or their legal guardian(s), and the study was previously approved by the Ethics Committee of the University of Santo Amaro, under numbers 4,350,476 and 6,048,872. This study respects Resolution No. 466/2012 of the National Health Council on Ethics in Research with Humans [67] and complies with the guidelines of the Declaration of Helsinki for Research with Humans [23]. Consent for publication Not applicable. Availability of data and materials The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Competing Interests The authors declare that they have no competing interests. Funding This study was supported by the São Paulo Research Foundation (FAPESP), grants #2019/14115-0 and 2022/02030-3, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) grant #305008/2022-6 (ALLB). Author Contribution FRM participated in the: Conceptualization, Development, Data Curation, Formal Analysis, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing. AN participated in the Collections and Laboratory Analysis. VP participated in the Collections and Laboratory Analysis. JBA participated in the Collections and Laboratory analysis. GRA participated in the Planning, Development, Clinical and Physical Evaluation. CAFS participated in the Planning, Development, Clinical and Physical Evaluation. GEF participated in Data Analysis and Statistics, Writing - review & editing. FR participated in Writing - review & editing. JBMS participated in the Development, Clinical and Physical Evaluation, Collections and Laboratory Analysis. MR participated in Data Analysis and Statistics. RVP participated in the Writing - review & editing. MST participated in the Writing - review & editing. CNF participated in the: Conceptualization, Development, Formal analysis, Validation, Writing - original draft, Writing - review & editing. ALLB participated in the: Conceptualization, Development, Data curation, Formal analysis, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing. All authors read and approved the final manuscript. Acknowledgments The authors are grateful to the older people who participated in the study and those who assisted with and cooperated in this study. Guilherme Furtado thank the National funding by FCT- Foundation for Science and Technology, P.I., through the institutional scientific employment program-contracts (CEECINST/00077/2021). Use of artificial intelligence The authors declare that any type of artificial intelligence did not automatically create this study. References Pezzullo AM, Axfors C, Contopoulos-Ioannidis DG, Apostolatos A, Ioannidis JPA. Age-stratified infection fatality rate of COVID-19 in the non-elderly population. Environ Res. 2023;216(Pt 3):114655. doi: 10.1016/j.envres.2022.114655. Dessie ZG, Zewotir T. Mortality-related risk factors of COVID-19: a systematic review and meta-analysis of 42 studies and 423,117 patients. BMC Infect Dis. 2021;21(1):855. doi: 10.1186/s12879-021-06536-3. Chen Y, Klein SL, Garibaldi BT, Li H, Wu C, Osevala NM, Li T, Margolick JB, Pawelec G, Leng SX. Aging in COVID-19: Vulnerability, immunity and intervention. Ageing Res Rev. 2021;65:101205. doi: 10.1016/j.arr.2020.101205. Witkowski JM, Fulop T, Bryl E. Immunosenescence and COVID-19. Mech Ageing Dev. 2022;204:111672. doi: 10.1016/j.mad.2022.111672. Linehan E, Fitzgerald DC. Ageing and the immune system: focus on macrophages. Eur J Microbiol Immunol (Bp). 2015;5(1):14-24. doi: 10.1556/EUJMI-D-14-00035 Eichhorn T, Huber S, Weiss R, Ebeyer-Masotta M, Lauková L, Emprechtinger R, et al. Infection with SARS-CoV-2 Is Associated with Elevated Levels of IP-10, MCP-1, and IL-13 in Sepsis Patients. Diagnostics (Basel). 2023;13(6):1069. doi: 10.3390/diagnostics13061069. Donlan AN, Sutherland TE, Marie C, Preissner S, Bradley BT, Carpenter RM, et al. IL-13 is a driver of COVID-19 severity. JCI Insight. 2021;6(15):e150107. doi: 10.1172/jci.insight.150107. Schulert GS, Grom AA. Pathogenesis of macrophage activation syndrome and potential for cytokine- directed therapies. Annu Rev Med. 2015;66:145-59. doi: 10.1146/annurev-med-061813-012806. de Carvalho DC, Fonseca FAH, Izar MCO, Silveira ALPA, Tuleta ID, do Amaral JB, et al. Monocytes presenting a pro-inflammatory profile persist in patients submitted to a long-term pharmacological treatment after acute myocardial infarction. Front Physiol. 2023;13:1056466. doi: 10.3389/fphys.2022.1056466. Zhou Y, Fu B, Zheng X, Wang D, Zhao C, Qi Y, et al. Pathogenic T-cells and inflammatory monocytes incite inflammatory storms in severe COVID-19 patients. Natl Sci Rev. 2020;7(6):998-1002. doi: 10.1093/nsr/nwaa041. Zou Y, Sun X, Wang Y, Ye X, Tu J, Yu R, et al. Integrating single-cell RNA sequencing data to genome-wide association analysis data identifies significant cell types in influenza A virus infection and COVID-19. Brief Funct Genomics. 2024;23(2):110-117. doi: 10.1093/bfgp/elad025. Ruenjaiman V, Sodsai P, Kueanjinda P, Bunrasmee W, Klinchanhom S, Reantragoon R, et al. Impact of SARS-CoV-2 infection on the profiles and responses of innate immune cells after recovery. J Microbiol Immunol Infect. 2022;55(6 Pt 1):993-1004. doi: 10.1016/j.jmii.2022.09.001. Ong SM, Teng K, Newell E, Chen H, Chen J, Loy T, et al. A Novel, Five-Marker Alternative to CD16-CD14 Gating to Identify the Three Human Monocyte Subsets. Front Immunol. 2019;10:1761. doi: 10.3389/fimmu.2019.01761. Gautier EL, Jakubzick C, Randolph GJ. Regulation of the migration and survival of monocyte subsets by chemokine receptors and its relevance to atherosclerosis. Arterioscler Thromb Vasc Biol. 2009;29(10):1412-8. doi: 10.1161/ATVBAHA.108.180505. Geissmann F, Jung S, Littman DR. Blood monocytes consist of two principal subsets with distinct migratory properties. Immunity. 2003;19(1):71-82. doi: 10.1016/s1074-7613(03)00174-2. Pence BD. Severe COVID-19 and aging: are monocytes the key? Geroscience. 2020;42(4):1051-1061. doi: 10.1007/s11357-020-00213-0. Albright JM, Dunn RC, Shults JA, Boe DM, Afshar M, Kovacs EJ. Advanced Age Alters Monocyte and Macrophage Responses. Antioxid Redox Signal. 2016;25(15):805-815. doi: 10.1089/ars.2016.6691. Aiello A, Ligotti ME, Garnica M, Accardi G, Calabrò A, Pojero F, et al. How Can We Improve Vaccination Response in Old People? Part I: Targeting Immunosenescence of Innate Immunity Cells. Int J Mol Sci. 2022;23(17):9880. doi: 10.3390/ijms23179880. Hadj Hassine I. Covid-19 vaccines and variants of concern: A review. Rev Med Virol. 2022;32(4):e2313. doi: 10.1002/rmv.2313. Crooke SN, Goergen KM, Ovsyannikova IG, Kennedy RB. Inflammasome Activity in Response to Influenza Vaccination Is Maintained in Monocyte-Derived Peripheral Blood Macrophages in Older Adults. Front Aging. 2021 20;2:719103. doi: 10.3389/fragi.2021.719103. Mohanty S, Joshi SR, Ueda I, Wilson J, Blevins TP, Siconolfi B, et al. Prolonged proinflammatory cytokine production in monocytes modulated by interleukin 10 after influenza vaccination in older adults. J Infect Dis. 2015;211(7):1174-84. doi: 10.1093/infdis/jiu573. Shen CF, Yen CL, Fu YC, Cheng CM, Shen TC, Chang PD, et al. Innate Immune Responses of Vaccinees Determine Early Neutralizing Antibody Production After ChAdOx1nCoV-19 Vaccination. Front Immunol. 2022;13:807454. doi: 10.3389/fimmu.2022.807454. World Medical Association. “World Medical Association Declaration of Helsinki. Ethical principles for medical research involving human subjects. Nurs Ethics. 2002;9(1):105-9. doi: 10.1191/0969733002ne486xx. Bachi AL, Suguri VM, Ramos LR, Mariano M, Vaisberg M, Lopes JD. Increased production of autoantibodies and specific antibodies in response to influenza virus vaccination in physically active older individuals. Results Immunol. 2013;3:10-6. doi: 10.1016/j.rinim.2013.01.001. Felismino ES, Santos JMB, Rossi M, Santos CAF, Durigon EL, Oliveira DBL, et al. Better Response to Influenza Virus Vaccination in Physically Trained Older Adults Is Associated With Reductions of Cytomegalovirus-Specific Immunoglobulins as Well as Improvements in the Inflammatory and CD8 + T-Cell Profiles. Front Immunol. 2021;12:713763. doi: 10.3389/fimmu.2021.713763. Dos Santos JMB, do Amaral JB, França CN, Monteiro FR, Alvares-Saraiva AM, Kalil S, et al. Distinct Immunological Profiles Help in the Maintenance of Salivary Secretory IgA Production in Mild Symptoms COVID-19 Patients. Front Immunol. 2022;13:890887. doi: 10.3389/fimmu.2022.890887. Silva BR, Monteiro FR, Cezário K, Amaral JBD, Paixão V, Almeida EB, et al. Older Adults Who Maintained a Regular Physical Exercise Routine before the Pandemic Show Better Immune Response to Vaccination for COVID-19. Int J Environ Res Public Health. 2023;20(3):1939. doi: 10.3390/ijerph20031939. Sauré D, O'Ryan M, Torres JP, Zuniga M, Santelices E, Basso LJ. Dynamic IgG seropositivity after rollout of CoronaVac and BNT162b2 COVID-19 vaccines in Chile: a sentinel surveillance study. Lancet Infect Dis. 2022;22(1):56-63. doi: 10.1016/S1473-3099(21)00479-5. Ranzani OT, Hitchings MDT, Dorion M, D'Agostini TL, de Paula RC, de Paula OFP, et al. Effectiveness of the CoronaVac vaccine in older adults during a gamma variant associated epidemic of covid-19 in Brazil: test negative case-control study. BMJ. 2021;374:n2015. doi: 10.1136/bmj.n2015. Erratum in: BMJ. 2021;374:n2091. doi: 10.1136/bmj.n2091. He Q, Mao Q, Zhang J, Bian L, Gao F, Wang J, et al. COVID-19 Vaccines: Current Understanding on Immunogenicity, Safety, and Further Considerations. Front Immunol. 2021;12:669339. doi: 10.3389/fimmu.2021.669339. Vitallé J, Pérez-Gómez A, Ostos FJ, Gasca-Capote C, Jiménez-León MR, Bachiller S, et al. Immune defects associated with lower SARS-CoV-2 BNT162b2 mRNA vaccine response in aged people. JCI Insight. 2022;7(17):e161045. doi: 10.1172/jci.insight.161045. Acosta-Altamirano G, Garduño-Javier E, Hernández-Gómez V, Espinosa JA, Vaca-Paniagua F, Rodríguez-Sosa M, et al. Dual activation profile of monocytes is associated with protection in Mexican patients during SARS-CoV-2 disease. Appl Microbiol Biotechnol. 2022;106(23):7905-7916. doi: 10.1007/s00253-022-12256-8. Antunes MDSM, Sugiyama FHC, Gravina HD, Castro RC, Mercado FJR, de Lima JO, et al. COVID-19 inactivated and non-replicating viral vector vaccines induce regulatory training phenotype in human monocytes under epigenetic control. Front Cell Infect Microbiol. 2023;13:1200789. doi: 10.3389/fcimb.2023.1200789. Shen CF, Yen CL, Fu YC, Cheng CM, Shen TC, Chang PD, et al. Innate Immune Responses of Vaccinees Determine Early Neutralizing Antibody Production After ChAdOx1nCoV-19 Vaccination. Front Immunol. 2022;13:807454. doi: 10.3389/fimmu.2022.807454. Murphy DM, Cox DJ, Connolly SA, Breen EP, Brugman AA, Phelan JJ, et al. Trained immunity is induced in humans after immunization with an adenoviral vector COVID-19 vaccine. J Clin Invest. 2023;133(2):e162581. doi: 10.1172/JCI162581. Cao C, Jiang J, Liu M, Dai Y, Chang T, Ji T, Gong F. Longitudinal evaluation of innate immune responses to three doses of CoronaVac vaccine. Front Immunol. 2023;14:1277831. doi: 10.3389/fimmu.2023.1277831. Teijaro JR, Farber DL. COVID-19 vaccines: modes of immune activation and future challenges. Nat Rev Immunol. 2021;21(4):195-197. doi: 10.1038/s41577-021-00526-x. García-Montero C, Fraile-Martínez O, Bravo C, Torres-Carranza D, Sanchez-Trujillo L, Gómez-Lahoz AM, et al. An Updated Review of SARS-CoV-2 Vaccines and the Importance of Effective Vaccination Programs in Pandemic Times. Vaccines (Basel). 2021;9(5):433. doi: 10.3390/vaccines9050433. Cao Y, Fan Y, Li F, Hao Y, Kong Y, Chen C, et al. Phenotypic and functional alterations of monocyte subsets with aging. Immun Ageing. 2022;19(1):63. doi: 10.1186/s12979-022-00321-9. Wang C, Yang S, Duan L, Du X, Tao J, Wang Y, et al. Adaptive immune responses and cytokine immune profiles in humans following prime and boost vaccination with the SARS-CoV-2 CoronaVac vaccine. Virol J. 2022;19(1):223. doi: 10.1186/s12985-022-01957-1. Santoro A, Bientinesi E, Monti D. Immunosenescence and inflammaging in the aging process: age-related diseases or longevity? Ageing Res Rev. 2021;71:101422. doi: 10.1016/j.arr.2021.101422. Santos CAF, Amirato GR, Paixão V, Almeida EB, Do Amaral JB, Monteiro FR, et al. Association among inflammaging, body composition, physical activity, and physical function tests in physically active women. Front Med (Lausanne). 2023;10:1206989. doi: 10.3389/fmed.2023.1206989. Rosseto-Welter EA, Rodrigues SS, de Figueiredo AB, França CN, Oliveira DBL, Bachi ALL, et al. Cellular and Humoral Immune Responses to Vaccination for COVID-19 Are Negatively Impacted by Senescent T Cells: A Case Report. Vaccines (Basel). 2023;11(4):840. doi: 10.3390/vaccines11040840. Zhao W, Chen W, Li J, Chen M, Li Q, Lv M, et al. Status of Humoral and Cellular Immune Responses within 12 Months following CoronaVac Vaccination against COVID-19. mBio. 2022;13(3):e0018122. doi: 10.1128/mbio.00181-22. Bilgin H, Marku M, Yilmaz SS, Karahasan Yagci A, Sili U, et al. The effect of immunization with inactivated SARS-CoV-2 vaccine (CoronaVac) and/or SARS-CoV-2 infection on antibody levels, plasmablasts, long-lived-plasma-cells, and IFN-γ release by natural killer cells. Vaccine. 2022;40(18):2619-2625. doi: 10.1016/j.vaccine.2022.03.001. Borriello F, Poli V, Shrock E, Spreafico R, Liu X, Pishesha N, et al. An adjuvant strategy enabled by modulation of the physical properties of microbial ligands expands antigen immunogenicity. Cell. 2022;185(4):614-629.e21. doi: 10.1016/j.cell.2022.01.009. Wu Z, Zheng Y, Sheng J, Han Y, Yang Y, Pan H, et al. CD3 + CD4 - CD8 - (Double-Negative) T Cells in Inflammation, Immune Disorders and Cancer. Front Immunol. 2022;13:816005. doi: 10.3389/fimmu.2022.816005. Oberhardt V, Luxenburger H, Kemming J, Schulien I, Ciminski K, Giese S, et al. Rapid and stable mobilization of CD8 + T cells by SARS-CoV-2 mRNA vaccine. Nature. 2021;597(7875):268-273. doi: 10.1038/s41586-021-03841-4. Valim V, Martins-Filho OA, Gouvea MDPG, Camacho LAB, Villela DAM, de Lima SMB, et al. Half Dose ChAdOx Study Group. Effectiveness, safety, and immunogenicity of half dose ChAdOx1 nCoV-19 COVID-19 Vaccine: Viana project. Front Immunol. 2022;13:966416. doi: 10.3389/fimmu.2022.966416. Tanaka T, Narazaki M, Masuda K, Kishimoto T. Regulation of IL-6 in Immunity and Diseases. Adv Exp Med Biol. 2016;941:79-88. doi: 10.1007/978-94-024-0921-5_4. Kang S, Narazaki M, Metwally H, Kishimoto T. Historical overview of the interleukin-6 family cytokine. J Exp Med. 2020;217(5):e20190347. doi: 10.1084/jem.20190347. Erratum in: J Exp Med. 2020;217(5):jem.2019034704212020c. doi: 10.1084/jem.2019034704212020c. Sekiya T, Mifsud EJ, Ohno M, Nomura N, Sasada M, Fujikura D, et al. Inactivated whole virus particle vaccine with potent immunogenicity and limited IL-6 induction is ideal for influenza. Vaccine. 2019;37(15):2158-2166. doi: 10.1016/j.vaccine.2019.02.057. Lindo J, Coelho P, Gavinhos C, Martins M, Liberal J, Ferreira AJ, Gonçalves T, Rodrigues F. Health Status and COVID-19 Epidemiology in an Inland Region of Portugal: A Retrospective Study. International Journal of Environmental Research and Public Health . 2024; 21(8):1033. https://doi.org/10.3390/ijerph21081033 . Wu C, Chen X, Cai Y, Xia J, Zhou X, Xu S, et al. Risk Factors Associated With Acute Respiratory Distress Syndrome and Death in Patients With Coronavirus Disease 2019 Pneumonia in Wuhan, China. JAMA Intern Med. 2020;180(7):934-943. doi: 10.1001/jamainternmed.2020.0994. Erratum in: JAMA Intern Med. 2020;180(7):1031. doi: 10.1001/jamainternmed.2020.1429. Jones SC, Brahmakshatriya V, Huston G, Dibble J, Swain SL. TLR-activated dendritic cells enhance the response of aged naive CD4 T cells via an IL-6-dependent mechanism. J Immunol. 2010;185(11):6783-94. doi: 10.4049/jimmunol.0901296. Zhou X, Hopkins JW, Wang C, Brahmakshatriya V, Swain SL, Kuchel GA, et al. IL-2 and IL-6 cooperate to enhance the generation of influenza-specific CD8 T cells responding to live influenza virus in aged mice and humans. Oncotarget. 2016;7(26):39171-39183. doi: 10.18632/oncotarget.10047. Romagnani S. T-cell subsets (Th1 versus Th2). Ann Allergy Asthma Immunol. 2000;85(1):9-18; quiz 18, 21. doi: 10.1016/S1081-1206(10)62426-X. Croft M. The TNF family in T cell differentiation and function--unanswered questions and future directions. Semin Immunol. 2014;26(3):183-90. doi: 10.1016/j.smim.2014.02.005. Jarczak D, Nierhaus A. Cytokine Storm-Definition, Causes, and Implications. Int J Mol Sci. 2022;23(19):11740. doi: 10.3390/ijms231911740. Liwsrisakun C, Pata S, Laopajon W, Takheaw N, Chaiwong W, Duangjit P,et al. Humoral and cellular immune responses against SARS-CoV-2 variants of concern induced by heterologous CoronaVac/ChAdOx-1 versus homologous ChAdOx-1 vaccination in the elderly. Asian Pac J Allergy Immunol. 2023. doi: 10.12932/AP-120822-1434. Totura AL, Baric RS. SARS coronavirus pathogenesis: host innate immune responses and viral antagonism of interferon. Curr Opin Virol. 2012;2(3):264-75. doi: 10.1016/j.coviro.2012.04.004. Dos Santos JMB, Soares CP, Monteiro FR, Mello R, do Amaral JB, Aguiar AS, et al. In Nasal Mucosal Secretions, Distinct IFN and IgA Responses Are Found in Severe and Mild SARS-CoV-2 Infection. Front Immunol. 2021;12:595343. doi: 10.3389/fimmu.2021.595343. Contoli M, Papi A, Tomassetti L, Rizzo P, Vieceli Dalla Sega F, Fortini F, et al. Blood Interferon-α Levels and Severity, Outcomes, and Inflammatory Profiles in Hospitalized COVID-19 Patients. Front Immunol. 2021;12:648004. doi: 10.3389/fimmu.2021.648004. Faure E, Poissy J, Goffard A, Fournier C, Kipnis E, Titecat M,et al. Distinct immune response in two MERS-CoV-infected patients: can we go from bench to bedside? PLoS One. 2014;9(2):e88716. doi: 10.1371/journal.pone.0088716. Mahallawi WH, Khabour OF, Zhang Q, Makhdoum HM, Suliman BA. MERS-CoV infection in humans is associated with a pro-inflammatory Th1 and Th17 cytokine profile. Cytokine. 2018;104:8-13. doi: 10.1016/j.cyto.2018.01.025. DeDiego ML, Nieto-Torres JL, Jimenez-Guardeño JM, Regla-Nava JA, Castaño-Rodriguez C, Fernandez-Delgado R, et al. Coronavirus virulence genes with main focus on SARS-CoV envelope gene. Virus Res. 2014;194:124-37. doi: 10.1016/j.virusres.2014.07.024. Novoa PC. What changes in research ethics in Brazil: resolution no. 466/12 of the National Health Council. Einstein (Sao Paulo). 2014;12(1):vii-x. doi: 10.1590/s1679-45082014ed3077. Additional Declarations No competing interests reported. Supplementary Files Highlights.pdf Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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2","display":"","copyAsset":false,"role":"figure","size":73809,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figure2Article.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7085263/v1/b37c376ec1b6e86ae3b94d80.jpg"},{"id":86751000,"identity":"22cb583b-d0a1-4c33-ad4b-a7b87a7bdc05","added_by":"auto","created_at":"2025-07-15 08:37:40","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":65506,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figure3Article.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7085263/v1/c8340c8db4cbd93dc1c2a592.jpg"},{"id":86750496,"identity":"3d358661-8ecc-4b6d-8579-aa1e93b466e8","added_by":"auto","created_at":"2025-07-15 08:29:40","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":85512,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figure4Article.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7085263/v1/316449051f42db745e95be58.jpg"},{"id":86752178,"identity":"3cc403e5-3a67-4686-a48c-6119208dc90d","added_by":"auto","created_at":"2025-07-15 08:45:40","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":157507,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figure5Article.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7085263/v1/0b9a19fc396fa75eb60ac441.jpg"},{"id":86752829,"identity":"b32098d6-4d62-49cf-b298-74d76be7e832","added_by":"auto","created_at":"2025-07-15 08:53:52","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":148643,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figure6Article.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7085263/v1/2df5e0099b812469b7fa52c9.jpg"},{"id":89562408,"identity":"f52d6c43-383f-4241-a1d8-6bcaf8460334","added_by":"auto","created_at":"2025-08-21 10:24:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1540643,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7085263/v1/65e532ea-83b9-4905-beab-5bbb1ec2d6e7.pdf"},{"id":86750999,"identity":"7ca91c09-f236-496c-b974-97e3fe073b5d","added_by":"auto","created_at":"2025-07-15 08:37:40","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":90998,"visible":true,"origin":"","legend":"","description":"","filename":"Highlights.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7085263/v1/58d0111ed9aff922076980b1.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Immunization with CoronaVac or ChadOx-1 Elicit Distinct Modulation of Monocyte Subtypes and Systemic Inflammatory Status in Older Adults","fulltext":[{"header":"Background","content":"\u003cp\u003eAmong some characteristics, the older adult population is one of the most affected groups by coronavirus disease 19 (COVID-19) [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e], and the increased susceptibility and severity in this population are attributed to immunosenescence [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e], a phenomenon associated with a gradual decline in immune cell functions, including monocytes [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eMonocytes are crucial in the inflammatory process and are classified into three subtypes: classical (CD14\u0026thinsp;+\u0026thinsp;+\u0026thinsp;CD16-), non-classical (CD14\u0026thinsp;+\u0026thinsp;CD16++), and intermediate (CD14\u0026thinsp;+\u0026thinsp;+\u0026thinsp;CD16+) [\u003cspan additionalcitationids=\"CR7 CR8 CR9 CR10 CR11 CR12 CR13 CR14\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Classical monocytes, comprising around 90% of total circulating monocytes, are considered inflammatory cells and produce several inflammatory cytokines [\u003cspan additionalcitationids=\"CR7 CR8\" citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Non-classical monocytes, constituting approximately 10% of circulating monocytes, exhibit an anti-inflammatory phenotype and are more responsive to virus-associated stimuli [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. Intermediate monocytes, representing up to 5% of circulating monocytes, display high phagocytic capacity and antigen presentation ability [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAging is associated with a tendency towards higher proportions of non-classical monocytes, although it was shown that monocytes from older adults exhibit increased basal cytokine production compared to those from younger adults. Additionally, non-classical monocytes display characteristics of cellular senescence, suggesting a link between aging and changes in monocyte subset proportions [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. It is important to mention that the raised expression of genes related to inflammation by monocytes of aged people drives the decreased vaccination responses, indicating a marked interplay between the elevation of the monocyte number and the inflammatory status, which can negatively impact vaccination responses [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eRegarding the COVID-19 vaccination in Brazil, it was available four types of vaccines with different mechanisms of action, such as the mRNA vaccine from Pfizer/BioNTech (BNT16b2) and Moderna (mRNA-1273), or non-recombinant viral vector vaccines from AstraZeneca/Oxford (ChAdOx-1, AZD1222) and Janssen-Cilag (Ad26.COV2.S), or whole-inactivated virus vaccine from Sinovac/Butant\u0026atilde; (CoronaVac). It is worth citing that among these, only CoronaVac does not specifically use the viral envelope glycoprotein called spike as the specific target [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAlthough monocytes play a pivotal role in increasing cytokine production and severity of COVID-19 in older adults, as well as in vaccination responses [\u003cspan additionalcitationids=\"CR17 CR18 CR19 CR20 CR21\" citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], the effects of vaccination with ChAdOx-1 and CoronaVac on monocyte subtypes in older adults remain unclear. Therefore, the present study aimed to assess these effects on monocyte subtypes and circulating cytokine levels.\u003c/p\u003e\u003cp\u003e\u003cb\u003eMethods\u003c/b\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003ePopulation and Design of the Study\u003c/b\u003e\u003c/p\u003e\u003cp\u003eIn this observational study, at two phases (pre- and post-vaccination), 66 older adults, aged between 60 and 85 years, of both sexes, and submitted to vaccination for SARS-CoV-2 were enrolled. All participants were recruited from the Older Adults Health Promotion Outpatient Clinic belonging to the Discipline of Geriatrics and Gerontology at the Federal University of S\u0026atilde;o Paulo (UNIFESP), and follow-up for the same geriatrician.\u003c/p\u003e\u003cp\u003eAs shown in Fig.\u0026nbsp;1, clinical and physical data, and blood samples were obtained between January and February 2021 (before the first round of vaccination) and between March and April 2021, which corresponds to 30 days after the second dose of the CoronaVac vaccine, or between July and August 2021, which corresponds to 30 days after the second dose of the ChAdOx-1 vaccine. The present study followed all the guidelines of the Declaration of Helsinki [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eVolunteers did not exhibit respiratory or any other symptoms during the study and received two COVID-19 vaccine doses (inclusion criteria). Participants who presented positive serology for HIV, frailty, sarcopenia, cancer, thrombosis, renal, liver, and neurological diseases, or reported use of corticosteroid and anti-inflammatory medication, or convalescent plasma therapy before and during the study, or even did not attendance at sample collection sessions were excluded.\u003c/p\u003e\u003cp\u003e\u003cb\u003eSample Size Calculation Groups\u003c/b\u003e\u003c/p\u003e\u003cp\u003eUsing G*Power software, sample size and statistical power were estimated for a Student's T-test with an effect size of 0.30, α level of 5%, and a statistical power of 0.95 for two groups with pre- and post-vaccination measurements. Based on previous studies [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e], a minimum of 55 individuals was considered after accounting for a 30% margin of losses or refusals. Initially, 92 older adults were invited and agreed to participate, but by inclusion/exclusion criteria, 81 volunteers were eligible. However, 15 volunteers were excluded because they did not attend sample collection. Therefore, data and samples from two groups: one immunized with CoronaVac (n\u0026thinsp;=\u0026thinsp;48) and the other with ChAdOx-1 (n\u0026thinsp;=\u0026thinsp;18) were used here.\u003c/p\u003e\u003cp\u003e\u003cb\u003eVaccination for COVID-19\u003c/b\u003e\u003c/p\u003e\u003cp\u003eIn according to the vaccination schedule initially proposed for the aged population by the National Health Surveillance Agency (ANVISA) of Brazil, in January and February 2021, particularly this population could be immunized exclusively with the vaccine CoronaVac, which was produced with the whole inactivated virus. After two weeks, it was also available the ChAdOx-1/AstraZeneca (AZD1222), which was a chimpanzee non-replicating adenovirus vector vaccine encoding the spike glycoprotein of SARS-CoV-2 into the host cells. All volunteers received two doses of the same vaccine, but the interval between the first and second dose for the vaccination with CoronaVac was around 30 days, whereas for the ChAdOx-1 was around 120 days.\u003c/p\u003e\u003cp\u003e\u003cb\u003eCollection of Blood Samples\u003c/b\u003e\u003c/p\u003e\u003cp\u003ePeripheral blood samples were collected before and 30 days after the second dose of COVID-19 vaccines (CoronaVac or ChAdOx-1) to obtain plasma and peripheral blood mononuclear cells (PBMC). Plasma aliquots were frozen at -80\u0026ordm;C for subsequent analysis of systemic immunoglobulin G (IgG) and pro- and anti-inflammatory cytokine concentrations. PBMC was isolated, storage and used as follows described.\u003c/p\u003e\u003cp\u003e\u003cb\u003eDetermination of specific immunoglobulin G (IgG) for SARS-CoV-2\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe systemic specific IgG levels for SARS-CoV-2 antigens were determined using the ELISA test in plasma samples previously storage, following the protocol described by our group [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e\u003cp\u003e\u003cb\u003eImmunophenotyping of monocyte subtypes\u003c/b\u003e\u003c/p\u003e\u003cp\u003eBlood samples were mixed with PBS and layered on tubes containing Histopaque. followed by centrifugation to obtain the PBMC. After, the PBMC was isolated, washed, mixed with a frozen medium, and stored in liquid nitrogen until their analysis.\u003c/p\u003e\u003cp\u003ePBMC cells were transferred to a Falcon\u0026reg; tube containing 10% PBS-SFB solution, followed by centrifugation and resuspension in 1% PBS-BSA. The cells were then incubated with monoclonal antibodies conjugated to fluorochromes (CD14-FITC and CD16-PE), along with isotype controls from BD Biosciences. After incubation, the cells were washed and resuspended in PBS, and monocyte subtypes were assessed through the flow cytometer FACSCalibur (Becton Dickinson Immunodiagnostic Systems, San Jos\u0026eacute;, CA, USA), acquiring a minimum of 10,000 events. Data analysis was performed using BD CellQuest Pro software (Becton Dickinson Immunodiagnostic Systems, San Jose, CA, USA). Figure\u0026nbsp;2 shows the study's strategy used to define monocyte populations and non-classical (CD14\u0026thinsp;+\u0026thinsp;CD16++), intermediate (CD14\u0026thinsp;+\u0026thinsp;+\u0026thinsp;CD16+), and classical (CD14\u0026thinsp;+\u0026thinsp;+\u0026thinsp;CD16-) subtypes.\u003c/p\u003e\u003cp\u003e\u003cb\u003eDetermination of Cytokines\u003c/b\u003e\u003c/p\u003e\u003cp\u003ePlasma concentrations of IL-4, IL-6, IL-10, IL-12p70, IL-13, TNF-α, IFN-α, IFN-β, and IFN-γ were determined using the LEGENDplex\u0026trade; bead-based multiplex assay (Biolegend, San Diego, CA, USA), following the manufacturer's instructions, with a linear range of 2.4 to 10,000 pg/mL and high correlation coefficients for standard curves.\u003c/p\u003e\u003cdiv id=\"Sec2\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eFirstly, both normality and homogeneity of each data were determined by the Shapiro-Wilks and Levene tests, respectively. Parametric data (demographic and anthropometrics) were represented as mean and standard deviation (X_\u0026plusmn;_SD) and were assessed by\u003c/p\u003e\u003cp\u003eStudent t-test, whereas non-parametric data (immune/inflammatory parameters) were represented as median and interquartile ranges, and the Wilcoxon test was used for intragroup comparisons, whilst the Kruskal-Wallis test with Dunn's post-hoc test was used for intergroup evaluations. Besides, Spearman\u0026acute;s rank correlation coefficient test was also performed.\u003c/p\u003e\u003cp\u003eFor all analyses, it was used the GraphPadPrism8 software (GraphPad Software, La Jolla, CA, USA) and set the α risk at 5% (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e presents demographic and anthropometric data of volunteer groups before and after separation in agreement with the type of vaccine received (CoronaVac or ChAdOx-1). No differences were observed in these parameters.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eAnthropometric data presented as a mean and standard deviation (X\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) of the volunteers participating in the current study.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e\u003cp\u003eVolunteers\u0026acute; Group\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eVariables\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eCoronaVac\u003c/p\u003e\u003cp\u003e(n\u0026thinsp;=\u0026thinsp;48)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eChAdOx-1\u003c/p\u003e\u003cp\u003e(n\u0026thinsp;=\u0026thinsp;18)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003ep-value\u003c/em\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAge (years)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e74.0\u0026thinsp;\u0026plusmn;\u0026thinsp;4.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e76.6\u0026thinsp;\u0026plusmn;\u0026thinsp;8.8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.0966\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHeight (meters)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e1.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.7490\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eWeight (kg)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e63.75\u0026thinsp;\u0026plusmn;\u0026thinsp;12.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e70.15\u0026thinsp;\u0026plusmn;\u0026thinsp;16.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.0941\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBMI (kg/m\u0026sup2;)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e25.56\u0026thinsp;\u0026plusmn;\u0026thinsp;4.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e27.72\u0026thinsp;\u0026plusmn;\u0026thinsp;5.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e\u003cp\u003e0.0894\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eFigure 3 illustrates the specific antibody (IgG) response to SARS-CoV-2 antigens, showing higher IgG levels post-vaccination than pre-vaccination values for both groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 for CoronaVac and ChAdOx-1). However, the ChAdOx-1 group exhibited higher IgG levels than the CoronaVac group (p\u0026thinsp;=\u0026thinsp;0.0001). Additionally, the immunogenicity found in the ChAdOx-1 group was 72.2% (13/18), whereas in the CoronaVac group was 39.6% (19/48). It is important to mention that the immunogenicity value was set considering an increase of, at least, two-fold in IgG levels at the post-vaccination moment in comparison to the pre-vaccination values.\u003c/p\u003e\u003cp\u003eFigure 4 presents the results, expressed in percentages, of monocytes subtypes. ChAdOx-1 group exhibited significantly increased percentages of all subtypes post-vaccination as compared to both pre-vaccination values (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) and the CoronaVac group (p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). No significant differences were observed in the CoronaVac group.\u003c/p\u003e\u003cp\u003eFigure 5 displays the data concerning the systemic cytokine levels assessed here [IL-4 (A), IL-6 (B), IL-13 (C), IL-10 (D), IL-12p70 (E), TNF-α (F), IFN-α (G), IFN-β (H), and IFN-γ (I)]. It was observed that the ChAdOx-1 group, at post-vaccination, showed lower levels of IL-4 (A), IL-13 (C), IL-10 (D), IL-12p70 (E), IFN-β (H), and IFN-γ (I) than pre-vaccination and CoronaVac group. Besides, post-vaccination IFN-γ levels (I) in the CoronaVac group were higher than pre-vaccination and the ChAdOx-1 group post-vaccination. No differences were observed in IL-6 and TNF-α.\u003c/p\u003e\u003cp\u003eRegarding the analysis of the ratio between pro-inflammatory cytokines and the anti-inflammatory IL-10 cytokine (Fig.\u0026nbsp;6), at post-vaccination, the ChAdOx-1 group demonstrated higher IL-6/IL-10 (B) and TNF-α/IL-10 (E) ratios, along with lower IL-12p70/IL-10 (C), IL-13/IL-10 (D), and IFN-β/IL-10 (G) ratios than pre-vaccination and CoronaVac group. Additionally, the ChAdOx-1 group exhibited a higher IFN-α/IL-10 (F) ratio and lower IFN-γ/IL-10 (H) ratio post-vaccination than the CoronaVac group. No difference was observed in the IL-4/IL-10 ratio (A).\u003c/p\u003e\u003cp\u003eCorrelation analyses, detailed in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, revealed significant positive correlations between various monocyte subtypes at pre- and post-vaccination occasions in both volunteer groups. Notably, the ChAdOx-1 group exhibited specific positive correlations, including intermediate and classic monocytes with the IL-6/IL-10 ratio pre-vaccination, and non-classical monocytes with this ratio post-vaccination. Conversely, the CoronaVac group showed significant positive correlations between all monocyte subtypes and the IL-13/IL-10 ratio post-vaccination. Additionally, both groups displayed significant positive associations between systemic IgG levels and all monocyte subtypes post-vaccination. Particularly at the pre-vaccination occasion, significant correlations were found between classical monocytes and IL-12p70, as well as between intermediate and classical monocytes with IFN-α, contrasting with significant negative correlations involving intermediate and classical monocytes with IL-13, IL-10, or IFN-γ.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eTable 2.\u0026nbsp;\u003c/strong\u003eSpearman\u0026apos;s correlation coefficient analysis between monocyte subtypes, total serum levels of specific IgG for SARS-CoV-2, and pro- and anti-inflammatory cytokines in volunteer groups vaccinated with CoronaVac or ChAdOx-1 before the first dose (pre) and 30 days after the administration of the second dose (post) of these vaccines.\u003c/p\u003e\n\u003ctable style=\"border: none;width:499.55pt;border-collapse:collapse;\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width:69.35pt;border:solid windowtext 1.0pt;border-left:none;padding: .75pt 9.15pt 0in 9.15pt;height:22.3pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003eParameters\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003ePRE\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:103.25pt;border:solid windowtext 1.0pt;border-left:none;padding:.75pt 9.15pt 0in 9.15pt;height:22.3pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003ePre-vaccination\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width:68.9pt;border:solid windowtext 1.0pt;border-left:none;padding:.75pt 9.15pt 0in 9.15pt;height:22.3pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003eParameters\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003eCoronaVac\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:101.8pt;border:solid windowtext 1.0pt;border-left:none;padding:.75pt 9.15pt 0in 9.15pt;height:22.3pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003ePost-vaccination\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width:65.0pt;border:solid windowtext 1.0pt;border-left:none;padding:.75pt 9.15pt 0in 9.15pt;height:22.3pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003eParameters\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003eChAdOx-1\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" style=\"width:91.25pt;border-top:solid windowtext 1.0pt;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:none;padding:.75pt 9.15pt 0in 9.15pt;height:22.3pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003ePost-vaccination\u003c/span\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:19.8pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003erho-velue\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:19.8pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003ep-value\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:19.8pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003erho-value\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:19.8pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003ep-value\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:19.8pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003erho-value\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:19.8pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cstrong\u003e\u003cem\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:black;'\u003ep-value\u003c/span\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIntermediate x IL-13\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e-0.2781\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0237\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003eIgG x Non-classical\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.3781\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.0080\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003eIgG x Non-classical\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.8493\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e\u0026lt;0.0001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIntermediate x IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e-0.3220\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0083\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003eIgG x Intermediate\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.3557\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.0130\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003eIgG x Intermediate\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.8493\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e\u0026lt;0.0001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIntermediate x IFN-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family:Symbol;color:blue;\"\u003ea\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.2680\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0295\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003eIgG x Classical\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.3517\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.0142\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003eIgG x Cl\u0026aacute;ssical\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.8493\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e\u0026lt;0.0001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIntermediate x IFN-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family:Symbol;color:blue;\"\u003eg\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e-0.2840\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0208\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003eNon-classical x IL-13/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.3277\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.0230\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003eNon-classical x IL-6/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.5550\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.0168\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIntermediate x IL-6/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.5139\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e\u0026lt;0.0001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003eIntermediate x IL-13/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:middle;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.3277\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.0230\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003eIntermediate x IL-6/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.5550\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.0168\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIntermediate x TNF-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family: Symbol;color:blue;\"\u003ea\u003c/span\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.3958\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0010\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003eIL-13/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.3277\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:red;'\u003e0.0230\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003eIL-6/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.5550\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:#00C000;'\u003e0.0168\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIntermediate x IFN-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family:Symbol;color:blue;\"\u003ea\u003c/span\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.3774\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0017\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border:none;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border:none;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIL-13\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e-0.3026\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0135\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border:none;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;vertical-align:bottom;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:51.75pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;'\u003e\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:65.0pt;padding:.75pt 9.15pt 0in 9.15pt;height: 31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:43.8pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:47.45pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"border:none;width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e-0.3437\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0047\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:50.05pt;padding:.9pt .9pt 0in .9pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:51.75pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:65.0pt;padding:.75pt 9.15pt 0in 9.15pt;height: 31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:43.8pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:47.45pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIL-12p70\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e-0.2489\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0438\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:50.05pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:51.75pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:65.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:43.8pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:47.45pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIFN-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family:Symbol;color:blue;\"\u003ea\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.2750\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0253\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:50.05pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:51.75pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:65.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:43.8pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:47.45pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIFN-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family:Symbol;color:blue;\"\u003eg\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e-0.2896\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0183\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:50.05pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:51.75pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:65.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:43.8pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:47.45pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eClassical x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003eIL-6/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.5253\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e\u0026lt;0.0001\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:50.05pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:51.75pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:65.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:43.8pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:47.45pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family: \"Arial\",sans-serif;color:blue;'\u003eClassical\u003c/span\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e\u0026nbsp;x TNF-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family: Symbol;color:blue;\"\u003ea\u003c/span\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.4161\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0005\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"border:none;width:68.9pt;border:none;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:50.05pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:51.75pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:65.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:43.8pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"border:none;width:47.45pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width:69.35pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family: \"Arial\",sans-serif;color:blue;'\u003eClassical\u003c/span\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e\u0026nbsp;x\u003c/span\u003e\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family: \"Arial\",sans-serif;color:blue;'\u003eIFN-\u003c/span\u003e\u003cspan style=\"font-size:11px;font-family:Symbol;color:blue;\"\u003ea\u003c/span\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e/IL-10\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:48.2pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.3877\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:55.05pt;border-top:none;border-left:none;border-bottom:solid windowtext 1.0pt;border-right:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;font-size:11.0pt;font-family:\"Calibri\",sans-serif;text-align:center;line-height:normal;'\u003e\u003cspan style='font-size:11px;font-family:\"Arial\",sans-serif;color:blue;'\u003e0.0012\u003c/span\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width:68.9pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt 9.15pt 0in 9.15pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width:50.05pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width:51.75pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width:65.0pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width:43.8pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd style=\"width:47.45pt;border:none;border-bottom:solid windowtext 1.0pt;padding:.75pt .75pt 0in .75pt;height:31.75pt;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n\u003c/table\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn general, our results demonstrated that immunization with the ChAdOx-1 and CoronaVac vaccines elicited significant differences in the immune/inflammatory responses in older adults. In fact, it was possible to evidence that the group vaccinated with ChAdOx-1 showed a considerable reduction in the levels of some cytokines, both pro- and anti-inflammatory properties, as well as an increase in the percentages of all monocyte subtypes as compared to the results found in the group vaccinated with CoronaVac. In addition, it was verified that volunteers in the ChAdOx-1 group presented higher specific levels of IgG to SARS-CoV-2 antigens than the CoronaVac group.\u003c/p\u003e\u003cp\u003eThis observation of higher IgG levels in the ChAdOx-1 group than in the CoronaVac group post-vaccination corroborates a previous study in which\u0026thinsp;\u0026gt;\u0026thinsp;60-year-old individuals vaccinated with CoronaVac presented lower antibody responses than younger individuals immunized with the same vaccine and individuals with the same age immunized with a vaccine containing mRNA for the S (spike) protein of SARS-CoV-2 [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Moreover, it was reported that vaccination efficacy 14 days or more after administration of the second dose decreased with increasing age, for example, dropping from 61.8% among individuals aged 70 to 74 years to 48.9% among individuals aged 75 to 79 years, and to 28.0% among individuals aged\u0026thinsp;\u0026ge;\u0026thinsp;80 years [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Likewise, our group also formerly showed that older adults immunized with two doses of the ChAdOx-1 vaccine presented a better antibody response than the group immunized with two doses of CoronaVac vaccine and also that lower IgG levels post-vaccination were associated with age in the CoronaVac group [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Additionally, the observation that the ChAdOx-1 group presented higher immunogenicity than the CoronaVac group agrees with the literature [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eBeyond these findings, the positive correlations between IgG levels and the monocyte subtype percentages in both volunteer groups can reinforce the role of monocytes in vaccine responses, including COVID-19 [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eAlthough it was reported a reduced non-classical monocyte frequency in COVID-19, after the dose of vaccination, these monocytes exhibited a dual profile, concurrently expressing both pro- and anti-inflammatory cytokines, suggesting that these monocytes can act in protection against viral infection [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Since some vaccines can trigger a memory-like response in monocytes, it is plausible to suggest that COVID-19 vaccination could induce the training of monocytes [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Particularly, people immunized with the ChAdOx-1 vaccine showed that the monocyte activation was related to not only neutralization antibody levels but also increased capacity for antigen presentation and a cytokine profile that could favor the development of adaptive immunity [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Concerning the CoronaVac vaccine, both a significant increase in classical and intermediate monocyte percentages and a decrease in non-classical monocyte frequency were verified post-vaccination with two doses, which, according to the authors, can supposedly indicate that an antiviral and adaptive response was elicited [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThese pieces of information can corroborate our findings that an enhancement in the percentage of all monocyte subtypes post-vaccination was positively associated with systemic IgG levels in both groups of volunteers. However, here, only the ChAdOx-1 group showed a significant increase in the monocyte subtype percentages post-vaccination. Even if we cannot affirm, this finding could be related to the technology/design used in the ChAdOx-1 vaccine, since it was a chimpanzee non-replicating adenovirus vector vaccine encoding the spike glycoprotein of SARS-CoV-2 into the host cells, thus stimulating the innate immune response to especially prime the CD4\u0026thinsp;+\u0026thinsp;T cells to differentiate into effector and memory cells that will help B cells to differentiate into antibody-secreting plasma cells. Differently, the CoronaVac vaccine was designed to contain the inactivated whole SARS-CoV-2 virus, which is not only a widely used technique but can also elicit protective immunity [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Based on these pieces of information, seemingly, the vaccine that uses an \"intracellular delivery\" design, such as ChadOx-1, can be more effective in eliciting the enhancement of the percentage of all monocyte subtypes than the CoronaVac in older adults.\u003c/p\u003e\u003cp\u003eParticularly the increased percentage of monocytes observed in older adults, in conjunction with the alteration of monocyte subsets towards intermediate and non-classical phenotypes verified during aging, could potentially correlate with elevated production of some cytokines, such as IL-6, IL-10, and TNF-α [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. These data corroborated the positive correlations found not only between all monocyte subtype percentages at pre-vaccination, but also between classical monocytes with circulating levels of IL-13, TNF-α, and IFN-α, as well as with the ratios IL-6/IL-10 and TNF-α/IL-10, in contrast to the negative correlations with the IL-10 and IFN-γ levels. Likewise, intermediate monocytes were positively correlated with IFN-a levels and with the ratios IL-6/IL-10, TNF-α/IL-10, and IFN-α/IL-10, and negatively correlated with IL-10, IL-13, and IFN-γ levels. Therefore, these findings allow us to suggest that systemic inflammatory status can modulate the monocyte subtype percentages in older adults.\u003c/p\u003e\u003cp\u003eInterestingly, post-vaccination, whereas the monocyte subtypes in the CoronaVac group were positively correlated with the IL-13/IL-10 ratio, in the ChAdOx-1 group this positive correlation was with the IL-6/IL-10 ratio. Concerning these findings, it is worth mentioning that it is well-known that IL-6 and IL-13, two cytokines related to the T help (Th)-2 profile, can promote B cell differentiation and subsequent antibody production [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eRegarding the triad - cytokines/aging/vaccination - as previously mentioned the development and progression of immunosenescence and inflammaging can impair the vaccination responses [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e] since an aberrant cytokine production can reduce the ability to process and present antigens to T cells, which negatively impacts the B cells' activation and antibody production [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Therefore, circulating cytokines profile assessment is pivotal to better characterize the immune/inflammatory responses elicited by vaccination, particularly in older adults.\u003c/p\u003e\u003cp\u003eDespite the isolated analysis of systemic cytokine levels providing important data, it is paramount to point out that the ratio analysis can offer an accurate measure concerning the balance between pro- and anti-inflammatory cytokines in several contexts, including in aging [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Similarly to those findings for monocytes, a distinct and interesting signature regarding the systemic cytokine profile was evidenced in the volunteer groups post-vaccination for COVID-19.\u003c/p\u003e\u003cp\u003eIn this sense, the isolated cytokine evaluations showed, in a general way, two aspects that aroused interest: (1) reduced levels of most cytokines assessed here in the ChAdOx-1 group, whereas (2) only IFN-γ levels were reduced in the CoronaVac group. This last finding corroborates previous studies in which the number of IFN-γ-producing cells, particularly T and NK cells, in response to the recombinant S protein from SARS-CoV-2, increased around 28 days after the administration of two doses of the CoronaVac vaccine [\u003cspan additionalcitationids=\"CR44\" citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Therefore, we can suggest that this vaccine was able to elicit IFN-γ production by the immune cells, which also impacted the higher IFN-γ/IL-10 ratio found in this volunteer group. Although IFN-γ is closely related to cellular immune responses, it has been cited that this cytokine can also enhance the humoral immune responses post-immunization, especially by improving both the capacity of immune cells to perform virus clearance and vaccination efficacy [\u003cspan additionalcitationids=\"CR47\" citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eConcerning the decrease in some cytokines found in the ChAdOx-1 group, this was an unexpected finding since had been reported an increase in cytokine levels post-vaccination, including in older adult individuals [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. Despite we cannot explain why the cytokine levels were lower post-ChAdOx-1 vaccination when assessed individually, the data obtained in the ratio analysis can help us understand these findings. In this respect, the significant reductions of IL-12p70, IL-13, and IFN-β levels could be responsible for the lower ratio between these cytokines with IL-10, whereas the maintenance of IL-6, TNF-α, and IFN-α levels can drive the higher ratio between them with IL-10 found in this group.\u003c/p\u003e\u003cp\u003eRegarding IL-6, it is a pro-inflammatory cytokine with pleiotropic actions in different contexts [\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e], including COVID-19 and vaccination [\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]. It is worth highlighting that the maintenance of IL-6 levels in the volunteer groups was a favorable result since higher IL-6 levels were associated with both an increased risk of death [\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e] and the development of cytokine storm in COVID-19 patients, as well as the increase of inflammaging, which, consequently, impairs the vaccination responses [\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]. Particularly in the context of aging, it was reported that IL-6 not only was involved in restoring the generation, memory, and responses of CD4\u0026thinsp;+\u0026thinsp;T cells, as well as the CD8\u0026thinsp;+\u0026thinsp;T cells' response to influenza virus infection from aged mice [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e], but also this cytokine has a pivotal as a growth factor and for the differentiation of B cells into antibody-producing cells.\u003csup\u003e52\u003c/sup\u003e Additionally, a limited IL-6 production was associated with a better immune response to influenza virus vaccination [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Taken together, these data allow us to suggest that the \"controlled\" increase of the IL-6/IL-10 ratio could favor the immune response to the ChAdOx-1 vaccine in older adults.\u003c/p\u003e\u003cp\u003eBeyond this finding, the higher ratios of TNF-α/IL-10 and IFN-α/IL-10 post-vaccination in the ChAdOx-1 group can demonstrate that a Th-1 immune profile was elicited. Concerning TNF-a, a classical pro-inflammatory cytokine, it is involved in cell-mediated immunity and phagocyte-dependent inflammation, particularly in virus infection [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e], and also when produced by CD8\u0026thinsp;+\u0026thinsp;T cells and NK cells (typical Th-1 cells), this molecule can help in controlling viral replication [\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]. Similarly to IL-6, TNF-α overproduction has been associated with worse outcomes in COVID-19 and the development of inflammaging and immunosenescence, which includes negative effects in B cell responses [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. Therefore, the maintenance of TNF-α levels post-vaccination is a prominent finding since can demonstrate that the inflammaging was not increased by the vaccination, and also the increased TNF-α/IL-10 ratio found in the ChAdOx-1 group allows us to suggest that a Th-1 immune response was preferentially towards, which could improve the virus clearance and, consequently, reduces the COVID-19 severity [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eCorroborating this suggestion, it was also observed a higher IFN-α/IL-10 ratio in the ChadOx-1 group. It is paramount to mention that the different types of interferon [Type-I (IFN-α and IFN-β); type II (IFN-γ); or type-III (IFN-λ)] are exclusively related to Th-1 immune responses and have a central role in viral infections not only by promoting the control of these pathogens but also by eliciting a robust adaptive immune response that drives to an efficient viral clearance [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. Regarding coronavirus infection, patients with SARS-CoV [\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e, \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e] and MERS [\u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e64\u003c/span\u003e] presented increased IFN-α levels, and it was suggested that this finding could be related to the improvement of not only the antigen presentation but also a robust antiviral response for these viral infections [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e, \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e] since this cytokine can promote the monocyte differentiation to dendritic cell, which are pivotal for the antigen presentation to CD4\u0026thinsp;+\u0026thinsp;T cells [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eLastly, it is worth highlighting that some limitations of this study were: (1) the discrepant number of volunteers in the CoronaVac (n\u0026thinsp;=\u0026thinsp;48) and ChadOx-1 (n\u0026thinsp;=\u0026thinsp;18) groups, however, as formerly cited, this difference was attributed to the fact that, although these were the first vaccines for COVID-19 available in Brazil, especially for the older adults population the CoronaVac was the main vaccine offered at the beginning of COVID-19 vaccination; (2) the lack of data concerning the activation levels of monocyte subtypes; (3) the lack of comparison of the immunophenotyping of T and NK lymphocytes, which could improve the findings of the present study, especially concerning systemic inflammatory status.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eTaking together, our findings showed that older adults immunized with ChAdOx-1 presented higher specific IgG levels for SARS-CoV-2 antigens than the older adult group immunized with CoronaVac, and this difference can putatively be associated with the distinct responses observed in the percentage of monocyte subtypes and systemic inflammatory status found in these volunteer groups.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all subjects and/or their legal guardian(s), and the study was previously approved by the Ethics Committee of the University of Santo Amaro, under numbers 4,350,476 and 6,048,872. This study respects Resolution No. 466/2012 of the National Health Council on Ethics in Research with Humans [67] and complies with the guidelines of the Declaration of Helsinki for Research with Humans [23].\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by the São Paulo Research Foundation (FAPESP), grants #2019/14115-0 and 2022/02030-3, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) grant #305008/2022-6 (ALLB).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contribution\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFRM participated in the: Conceptualization, Development, Data Curation, Formal Analysis, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review \u0026amp; editing. AN participated in the Collections and Laboratory Analysis. VP participated in the Collections and Laboratory Analysis. JBA participated in the Collections and Laboratory analysis. GRA participated in the Planning, Development, Clinical and Physical Evaluation. CAFS participated in the Planning, Development, Clinical and Physical Evaluation. GEF participated in Data Analysis and Statistics, Writing - review \u0026amp; editing. FR participated in Writing - review \u0026amp; editing. JBMS participated in the Development, Clinical and Physical Evaluation, Collections and Laboratory Analysis. MR participated in Data Analysis and Statistics. RVP participated in the Writing - review \u0026amp; editing. MST participated in the Writing - review \u0026amp; editing. CNF participated in the: Conceptualization, Development, Formal analysis, Validation, Writing - original draft, Writing - review \u0026amp; editing. ALLB participated in the: Conceptualization, Development, Data curation, Formal analysis, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review \u0026amp; editing. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful to the older people who participated in the study and those who assisted with and cooperated in this study. Guilherme Furtado thank the National funding by FCT- Foundation for Science and Technology, P.I., through the institutional scientific employment program-contracts (CEECINST/00077/2021).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eUse of artificial intelligence\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that any type of artificial intelligence did not automatically create this study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003ePezzullo AM, Axfors C, Contopoulos-Ioannidis DG, Apostolatos A, Ioannidis JPA. Age-stratified infection fatality rate of COVID-19 in the non-elderly population. Environ Res. 2023;216(Pt 3):114655. doi: 10.1016/j.envres.2022.114655.\u003c/li\u003e\n\u003cli\u003eDessie ZG, Zewotir T. Mortality-related risk factors of COVID-19: a systematic review and meta-analysis of 42 studies and 423,117 patients. BMC Infect Dis. 2021;21(1):855. doi: 10.1186/s12879-021-06536-3.\u003c/li\u003e\n\u003cli\u003eChen Y, Klein SL, Garibaldi BT, Li H, Wu C, Osevala NM, Li T, Margolick JB, Pawelec G, Leng SX. Aging in COVID-19: Vulnerability, immunity and intervention. Ageing Res Rev. 2021;65:101205. doi: 10.1016/j.arr.2020.101205.\u003c/li\u003e\n\u003cli\u003eWitkowski JM, Fulop T, Bryl E. Immunosenescence and COVID-19. Mech Ageing Dev. 2022;204:111672. doi: 10.1016/j.mad.2022.111672.\u003c/li\u003e\n\u003cli\u003eLinehan E, Fitzgerald DC. Ageing and the immune system: focus on macrophages. Eur J Microbiol Immunol (Bp). 2015;5(1):14-24. doi: 10.1556/EUJMI-D-14-00035\u003c/li\u003e\n\u003cli\u003eEichhorn T, Huber S, Weiss R, Ebeyer-Masotta M, Laukov\u0026aacute; L, Emprechtinger R, et al. Infection with SARS-CoV-2 Is Associated with Elevated Levels of IP-10, MCP-1, and IL-13 in Sepsis Patients. Diagnostics (Basel). 2023;13(6):1069. doi: 10.3390/diagnostics13061069.\u003c/li\u003e\n\u003cli\u003eDonlan AN, Sutherland TE, Marie C, Preissner S, Bradley BT, Carpenter RM, et al. IL-13 is a driver of COVID-19 severity. JCI Insight. 2021;6(15):e150107. doi: 10.1172/jci.insight.150107.\u003c/li\u003e\n\u003cli\u003eSchulert GS, Grom AA. Pathogenesis of macrophage activation syndrome and potential for cytokine- directed therapies. Annu Rev Med. 2015;66:145-59. doi: 10.1146/annurev-med-061813-012806. \u003c/li\u003e\n\u003cli\u003ede Carvalho DC, Fonseca FAH, Izar MCO, Silveira ALPA, Tuleta ID, do Amaral JB, et al. Monocytes presenting a pro-inflammatory profile persist in patients submitted to a long-term pharmacological treatment after acute myocardial infarction. Front Physiol. 2023;13:1056466. doi: 10.3389/fphys.2022.1056466.\u003c/li\u003e\n\u003cli\u003eZhou Y, Fu B, Zheng X, Wang D, Zhao C, Qi Y, et al. Pathogenic T-cells and inflammatory monocytes incite inflammatory storms in severe COVID-19 patients. Natl Sci Rev. 2020;7(6):998-1002. doi: 10.1093/nsr/nwaa041.\u003c/li\u003e\n\u003cli\u003eZou Y, Sun X, Wang Y, Ye X, Tu J, Yu R, et al. Integrating single-cell RNA sequencing data to genome-wide association analysis data identifies significant cell types in influenza A virus infection and COVID-19. Brief Funct Genomics. 2024;23(2):110-117. doi: 10.1093/bfgp/elad025.\u003c/li\u003e\n\u003cli\u003eRuenjaiman V, Sodsai P, Kueanjinda P, Bunrasmee W, Klinchanhom S, Reantragoon R, et al. Impact of SARS-CoV-2 infection on the profiles and responses of innate immune cells after recovery. J Microbiol Immunol Infect. 2022;55(6 Pt 1):993-1004. doi: 10.1016/j.jmii.2022.09.001.\u003c/li\u003e\n\u003cli\u003eOng SM, Teng K, Newell E, Chen H, Chen J, Loy T, et al. A Novel, Five-Marker Alternative to CD16-CD14 Gating to Identify the Three Human Monocyte Subsets. Front Immunol. 2019;10:1761. doi: 10.3389/fimmu.2019.01761.\u003c/li\u003e\n\u003cli\u003eGautier EL, Jakubzick C, Randolph GJ. Regulation of the migration and survival of monocyte subsets by chemokine receptors and its relevance to atherosclerosis. Arterioscler Thromb Vasc Biol. 2009;29(10):1412-8. doi: 10.1161/ATVBAHA.108.180505.\u003c/li\u003e\n\u003cli\u003eGeissmann F, Jung S, Littman DR. Blood monocytes consist of two principal subsets with distinct migratory properties. Immunity. 2003;19(1):71-82. doi: 10.1016/s1074-7613(03)00174-2.\u003c/li\u003e\n\u003cli\u003ePence BD. Severe COVID-19 and aging: are monocytes the key? Geroscience. 2020;42(4):1051-1061. doi: 10.1007/s11357-020-00213-0.\u003c/li\u003e\n\u003cli\u003eAlbright JM, Dunn RC, Shults JA, Boe DM, Afshar M, Kovacs EJ. Advanced Age Alters Monocyte and Macrophage Responses. Antioxid Redox Signal. 2016;25(15):805-815. doi: 10.1089/ars.2016.6691.\u003c/li\u003e\n\u003cli\u003eAiello A, Ligotti ME, Garnica M, Accardi G, Calabr\u0026ograve; A, Pojero F, et al. How Can We Improve Vaccination Response in Old People? Part I: Targeting Immunosenescence of Innate Immunity Cells. Int J Mol Sci. 2022;23(17):9880. doi: 10.3390/ijms23179880.\u003c/li\u003e\n\u003cli\u003eHadj Hassine I. Covid-19 vaccines and variants of concern: A review. Rev Med Virol. 2022;32(4):e2313. doi: 10.1002/rmv.2313.\u003c/li\u003e\n\u003cli\u003eCrooke SN, Goergen KM, Ovsyannikova IG, Kennedy RB. Inflammasome Activity in Response to Influenza Vaccination Is Maintained in Monocyte-Derived Peripheral Blood Macrophages in Older Adults. Front Aging. 2021 20;2:719103. doi: 10.3389/fragi.2021.719103.\u003c/li\u003e\n\u003cli\u003eMohanty S, Joshi SR, Ueda I, Wilson J, Blevins TP, Siconolfi B, et al. Prolonged proinflammatory cytokine production in monocytes modulated by interleukin 10 after influenza vaccination in older adults. J Infect Dis. 2015;211(7):1174-84. doi: 10.1093/infdis/jiu573. \u003c/li\u003e\n\u003cli\u003eShen CF, Yen CL, Fu YC, Cheng CM, Shen TC, Chang PD, et al. Innate Immune Responses of Vaccinees Determine Early Neutralizing Antibody Production After ChAdOx1nCoV-19 Vaccination. Front Immunol. 2022;13:807454. doi: 10.3389/fimmu.2022.807454.\u003c/li\u003e\n\u003cli\u003eWorld Medical Association. \u0026ldquo;World Medical Association Declaration of Helsinki. Ethical principles for medical research involving human subjects. Nurs Ethics. 2002;9(1):105-9. doi: 10.1191/0969733002ne486xx.\u003c/li\u003e\n\u003cli\u003eBachi AL, Suguri VM, Ramos LR, Mariano M, Vaisberg M, Lopes JD. Increased production of autoantibodies and specific antibodies in response to influenza virus vaccination in physically active older individuals. Results Immunol. 2013;3:10-6. doi: 10.1016/j.rinim.2013.01.001. \u003c/li\u003e\n\u003cli\u003eFelismino ES, Santos JMB, Rossi M, Santos CAF, Durigon EL, Oliveira DBL, et al. Better Response to Influenza Virus Vaccination in Physically Trained Older Adults Is Associated With Reductions of Cytomegalovirus-Specific Immunoglobulins as Well as Improvements in the Inflammatory and CD8\u003csup\u003e+\u003c/sup\u003e T-Cell Profiles. Front Immunol. 2021;12:713763. doi: 10.3389/fimmu.2021.713763.\u003c/li\u003e\n\u003cli\u003eDos Santos JMB, do Amaral JB, Fran\u0026ccedil;a CN, Monteiro FR, Alvares-Saraiva AM, Kalil S, et al. Distinct Immunological Profiles Help in the Maintenance of Salivary Secretory IgA Production in Mild Symptoms COVID-19 Patients. Front Immunol. 2022;13:890887. doi: 10.3389/fimmu.2022.890887.\u003c/li\u003e\n\u003cli\u003eSilva BR, Monteiro FR, Cez\u0026aacute;rio K, Amaral JBD, Paix\u0026atilde;o V, Almeida EB, et al. Older Adults Who Maintained a Regular Physical Exercise Routine before the Pandemic Show Better Immune Response to Vaccination for COVID-19. Int J Environ Res Public Health. 2023;20(3):1939. doi: 10.3390/ijerph20031939.\u003c/li\u003e\n\u003cli\u003eSaur\u0026eacute; D, O\u0026apos;Ryan M, Torres JP, Zuniga M, Santelices E, Basso LJ. Dynamic IgG seropositivity after rollout of CoronaVac and BNT162b2 COVID-19 vaccines in Chile: a sentinel surveillance study. Lancet Infect Dis. 2022;22(1):56-63. doi: 10.1016/S1473-3099(21)00479-5.\u003c/li\u003e\n\u003cli\u003eRanzani OT, Hitchings MDT, Dorion M, D\u0026apos;Agostini TL, de Paula RC, de Paula OFP, et al. Effectiveness of the CoronaVac vaccine in older adults during a gamma variant associated epidemic of covid-19 in Brazil: test negative case-control study. BMJ. 2021;374:n2015. doi: 10.1136/bmj.n2015. Erratum in: BMJ. 2021;374:n2091. doi: 10.1136/bmj.n2091.\u003c/li\u003e\n\u003cli\u003eHe Q, Mao Q, Zhang J, Bian L, Gao F, Wang J, et al. COVID-19 Vaccines: Current Understanding on Immunogenicity, Safety, and Further Considerations. Front Immunol. 2021;12:669339. doi: 10.3389/fimmu.2021.669339.\u003c/li\u003e\n\u003cli\u003eVitall\u0026eacute; J, P\u0026eacute;rez-G\u0026oacute;mez A, Ostos FJ, Gasca-Capote C, Jim\u0026eacute;nez-Le\u0026oacute;n MR, Bachiller S, et al. Immune defects associated with lower SARS-CoV-2 BNT162b2 mRNA vaccine response in aged people. JCI Insight. 2022;7(17):e161045. doi: 10.1172/jci.insight.161045.\u003c/li\u003e\n\u003cli\u003eAcosta-Altamirano G, Gardu\u0026ntilde;o-Javier E, Hern\u0026aacute;ndez-G\u0026oacute;mez V, Espinosa JA, Vaca-Paniagua F, Rodr\u0026iacute;guez-Sosa M, et al. Dual activation profile of monocytes is associated with protection in Mexican patients during SARS-CoV-2 disease. Appl Microbiol Biotechnol. 2022;106(23):7905-7916. doi: 10.1007/s00253-022-12256-8.\u003c/li\u003e\n\u003cli\u003eAntunes MDSM, Sugiyama FHC, Gravina HD, Castro RC, Mercado FJR, de Lima JO, et al. COVID-19 inactivated and non-replicating viral vector vaccines induce regulatory training phenotype in human monocytes under epigenetic control. Front Cell Infect Microbiol. 2023;13:1200789. doi: 10.3389/fcimb.2023.1200789.\u003c/li\u003e\n\u003cli\u003eShen CF, Yen CL, Fu YC, Cheng CM, Shen TC, Chang PD, et al. Innate Immune Responses of Vaccinees Determine Early Neutralizing Antibody Production After ChAdOx1nCoV-19 Vaccination. Front Immunol. 2022;13:807454. doi: 10.3389/fimmu.2022.807454.\u003c/li\u003e\n\u003cli\u003eMurphy DM, Cox DJ, Connolly SA, Breen EP, Brugman AA, Phelan JJ, et al. Trained immunity is induced in humans after immunization with an adenoviral vector COVID-19 vaccine. J Clin Invest. 2023;133(2):e162581. doi: 10.1172/JCI162581.\u003c/li\u003e\n\u003cli\u003eCao C, Jiang J, Liu M, Dai Y, Chang T, Ji T, Gong F. Longitudinal evaluation of innate immune responses to three doses of CoronaVac vaccine. Front Immunol. 2023;14:1277831. doi: 10.3389/fimmu.2023.1277831.\u003c/li\u003e\n\u003cli\u003eTeijaro JR, Farber DL. COVID-19 vaccines: modes of immune activation and future challenges. Nat Rev Immunol. 2021;21(4):195-197. doi: 10.1038/s41577-021-00526-x.\u003c/li\u003e\n\u003cli\u003eGarc\u0026iacute;a-Montero C, Fraile-Mart\u0026iacute;nez O, Bravo C, Torres-Carranza D, Sanchez-Trujillo L, G\u0026oacute;mez-Lahoz AM, et al. An Updated Review of SARS-CoV-2 Vaccines and the Importance of Effective Vaccination Programs in Pandemic Times. Vaccines (Basel). 2021;9(5):433. doi: 10.3390/vaccines9050433. \u003c/li\u003e\n\u003cli\u003eCao Y, Fan Y, Li F, Hao Y, Kong Y, Chen C, et al. Phenotypic and functional alterations of monocyte subsets with aging. Immun Ageing. 2022;19(1):63. doi: 10.1186/s12979-022-00321-9.\u003c/li\u003e\n\u003cli\u003eWang C, Yang S, Duan L, Du X, Tao J, Wang Y, et al. Adaptive immune responses and cytokine immune profiles in humans following prime and boost vaccination with the SARS-CoV-2 CoronaVac vaccine. Virol J. 2022;19(1):223. doi: 10.1186/s12985-022-01957-1.\u003c/li\u003e\n\u003cli\u003eSantoro A, Bientinesi E, Monti D. Immunosenescence and inflammaging in the aging process: age-related diseases or longevity? Ageing Res Rev. 2021;71:101422. doi: 10.1016/j.arr.2021.101422.\u003c/li\u003e\n\u003cli\u003eSantos CAF, Amirato GR, Paix\u0026atilde;o V, Almeida EB, Do Amaral JB, Monteiro FR, et al. Association among inflammaging, body composition, physical activity, and physical function tests in physically active women. Front Med (Lausanne). 2023;10:1206989. doi: 10.3389/fmed.2023.1206989.\u003c/li\u003e\n\u003cli\u003eRosseto-Welter EA, Rodrigues SS, de Figueiredo AB, Fran\u0026ccedil;a CN, Oliveira DBL, Bachi ALL, et al. Cellular and Humoral Immune Responses to Vaccination for COVID-19 Are Negatively Impacted by Senescent T Cells: A Case Report. Vaccines (Basel). 2023;11(4):840. doi: 10.3390/vaccines11040840.\u003c/li\u003e\n\u003cli\u003eZhao W, Chen W, Li J, Chen M, Li Q, Lv M, et al. Status of Humoral and Cellular Immune Responses within 12 Months following CoronaVac Vaccination against COVID-19. mBio. 2022;13(3):e0018122. doi: 10.1128/mbio.00181-22.\u003c/li\u003e\n\u003cli\u003eBilgin H, Marku M, Yilmaz SS, Karahasan Yagci A, Sili U, et al. The effect of immunization with inactivated SARS-CoV-2 vaccine (CoronaVac) and/or SARS-CoV-2 infection on antibody levels, plasmablasts, long-lived-plasma-cells, and IFN-\u0026gamma; release by natural killer cells. Vaccine. 2022;40(18):2619-2625. doi: 10.1016/j.vaccine.2022.03.001.\u003c/li\u003e\n\u003cli\u003eBorriello F, Poli V, Shrock E, Spreafico R, Liu X, Pishesha N, et al. An adjuvant strategy enabled by modulation of the physical properties of microbial ligands expands antigen immunogenicity. Cell. 2022;185(4):614-629.e21. doi: 10.1016/j.cell.2022.01.009.\u003c/li\u003e\n\u003cli\u003eWu Z, Zheng Y, Sheng J, Han Y, Yang Y, Pan H, et al. CD3\u003csup\u003e+\u003c/sup\u003eCD4\u003csup\u003e-\u003c/sup\u003eCD8\u003csup\u003e-\u003c/sup\u003e (Double-Negative) T Cells in Inflammation, Immune Disorders and Cancer. Front Immunol. 2022;13:816005. doi: 10.3389/fimmu.2022.816005.\u003c/li\u003e\n\u003cli\u003eOberhardt V, Luxenburger H, Kemming J, Schulien I, Ciminski K, Giese S, et al. Rapid and stable mobilization of CD8\u003csup\u003e+\u003c/sup\u003e T cells by SARS-CoV-2 mRNA vaccine. Nature. 2021;597(7875):268-273. doi: 10.1038/s41586-021-03841-4.\u003c/li\u003e\n\u003cli\u003eValim V, Martins-Filho OA, Gouvea MDPG, Camacho LAB, Villela DAM, de Lima SMB, et al. Half Dose ChAdOx Study Group. Effectiveness, safety, and immunogenicity of half dose ChAdOx1 nCoV-19 COVID-19 Vaccine: Viana project. Front Immunol. 2022;13:966416. doi: 10.3389/fimmu.2022.966416.\u003c/li\u003e\n\u003cli\u003eTanaka T, Narazaki M, Masuda K, Kishimoto T. Regulation of IL-6 in Immunity and Diseases. Adv Exp Med Biol. 2016;941:79-88. doi: 10.1007/978-94-024-0921-5_4.\u003c/li\u003e\n\u003cli\u003eKang S, Narazaki M, Metwally H, Kishimoto T. Historical overview of the interleukin-6 family cytokine. J Exp Med. 2020;217(5):e20190347. doi: 10.1084/jem.20190347. Erratum in: J Exp Med. 2020;217(5):jem.2019034704212020c. doi: 10.1084/jem.2019034704212020c.\u003c/li\u003e\n\u003cli\u003eSekiya T, Mifsud EJ, Ohno M, Nomura N, Sasada M, Fujikura D, et al. Inactivated whole virus particle vaccine with potent immunogenicity and limited IL-6 induction is ideal for influenza. Vaccine. 2019;37(15):2158-2166. doi: 10.1016/j.vaccine.2019.02.057.\u003c/li\u003e\n\u003cli\u003eLindo J, Coelho P, Gavinhos C, Martins M, Liberal J, Ferreira AJ, Gon\u0026ccedil;alves T, Rodrigues F. Health Status and COVID-19 Epidemiology in an Inland Region of Portugal: A Retrospective Study. \u003cem\u003eInternational Journal of Environmental Research and Public Health\u003c/em\u003e. 2024; 21(8):1033. https://doi.org/10.3390/ijerph21081033 .\u003c/li\u003e\n\u003cli\u003eWu C, Chen X, Cai Y, Xia J, Zhou X, Xu S, et al. Risk Factors Associated With Acute Respiratory Distress Syndrome and Death in Patients With Coronavirus Disease 2019 Pneumonia in Wuhan, China. JAMA Intern Med. 2020;180(7):934-943. doi: 10.1001/jamainternmed.2020.0994. Erratum in: JAMA Intern Med. 2020;180(7):1031. doi: 10.1001/jamainternmed.2020.1429.\u003c/li\u003e\n\u003cli\u003eJones SC, Brahmakshatriya V, Huston G, Dibble J, Swain SL. TLR-activated dendritic cells enhance the response of aged naive CD4 T cells via an IL-6-dependent mechanism. J Immunol. 2010;185(11):6783-94. doi: 10.4049/jimmunol.0901296.\u003c/li\u003e\n\u003cli\u003eZhou X, Hopkins JW, Wang C, Brahmakshatriya V, Swain SL, Kuchel GA, et al. IL-2 and IL-6 cooperate to enhance the generation of influenza-specific CD8 T cells responding to live influenza virus in aged mice and humans. Oncotarget. 2016;7(26):39171-39183. doi: 10.18632/oncotarget.10047.\u003c/li\u003e\n\u003cli\u003eRomagnani S. T-cell subsets (Th1 versus Th2). Ann Allergy Asthma Immunol. 2000;85(1):9-18; quiz 18, 21. doi: 10.1016/S1081-1206(10)62426-X.\u003c/li\u003e\n\u003cli\u003eCroft M. The TNF family in T cell differentiation and function--unanswered questions and future directions. Semin Immunol. 2014;26(3):183-90. doi: 10.1016/j.smim.2014.02.005.\u003c/li\u003e\n\u003cli\u003eJarczak D, Nierhaus A. Cytokine Storm-Definition, Causes, and Implications. Int J Mol Sci. 2022;23(19):11740. doi: 10.3390/ijms231911740.\u003c/li\u003e\n\u003cli\u003eLiwsrisakun C, Pata S, Laopajon W, Takheaw N, Chaiwong W, Duangjit P,et al. Humoral and cellular immune responses against SARS-CoV-2 variants of concern induced by heterologous CoronaVac/ChAdOx-1 versus homologous ChAdOx-1 vaccination in the elderly. Asian Pac J Allergy Immunol. 2023. doi: 10.12932/AP-120822-1434.\u003c/li\u003e\n\u003cli\u003eTotura AL, Baric RS. SARS coronavirus pathogenesis: host innate immune responses and viral antagonism of interferon. Curr Opin Virol. 2012;2(3):264-75. doi: 10.1016/j.coviro.2012.04.004.\u003c/li\u003e\n\u003cli\u003eDos Santos JMB, Soares CP, Monteiro FR, Mello R, do Amaral JB, Aguiar AS, et al. In Nasal Mucosal Secretions, Distinct IFN and IgA Responses Are Found in Severe and Mild SARS-CoV-2 Infection. Front Immunol. 2021;12:595343. doi: 10.3389/fimmu.2021.595343.\u003c/li\u003e\n\u003cli\u003eContoli M, Papi A, Tomassetti L, Rizzo P, Vieceli Dalla Sega F, Fortini F, et al. Blood Interferon-\u0026alpha; Levels and Severity, Outcomes, and Inflammatory Profiles in Hospitalized COVID-19 Patients. Front Immunol. 2021;12:648004. doi: 10.3389/fimmu.2021.648004.\u003c/li\u003e\n\u003cli\u003eFaure E, Poissy J, Goffard A, Fournier C, Kipnis E, Titecat M,et al. Distinct immune response in two MERS-CoV-infected patients: can we go from bench to bedside? PLoS One. 2014;9(2):e88716. doi: 10.1371/journal.pone.0088716.\u003c/li\u003e\n\u003cli\u003eMahallawi WH, Khabour OF, Zhang Q, Makhdoum HM, Suliman BA. MERS-CoV infection in humans is associated with a pro-inflammatory Th1 and Th17 cytokine profile. Cytokine. 2018;104:8-13. doi: 10.1016/j.cyto.2018.01.025.\u003c/li\u003e\n\u003cli\u003eDeDiego ML, Nieto-Torres JL, Jimenez-Guarde\u0026ntilde;o JM, Regla-Nava JA, Casta\u0026ntilde;o-Rodriguez C, Fernandez-Delgado R, et al. Coronavirus virulence genes with main focus on SARS-CoV envelope gene. Virus Res. 2014;194:124-37. doi: 10.1016/j.virusres.2014.07.024.\u003c/li\u003e\n\u003cli\u003eNovoa PC. What changes in research ethics in Brazil: resolution no. 466/12 of the National Health Council. Einstein (Sao Paulo). 2014;12(1):vii-x. doi: 10.1590/s1679-45082014ed3077.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"ChAdOx-1, CoronaVac, cytokines, IgG, inflammation, SARS-CoV-2","lastPublishedDoi":"10.21203/rs.3.rs-7085263/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7085263/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cb\u003eBackground\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe older adult population was significantly affected by COVID-19 due to immunosenescence, which alters immune elements such as monocytes, impacting immune responses to pathogens and vaccination. Aging-associated systemic inflammation, or inflammaging, can further impair vaccine responses. Although important data exist, the effects of COVID-19 vaccination on monocytes and systemic inflammation in older adults are not fully understood. This study aimed to investigate the impact of COVID-19 vaccination with CoronaVac and ChAdOx-1 on monocyte subtypes and systemic inflammatory status in older adults.\u003c/p\u003e\u003cp\u003e\u003cb\u003eMethods\u003c/b\u003e\u003c/p\u003e\u003cp\u003eSixty-six older adults (74.74\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9 years) participated in this study. Blood samples were collected before and 30 days after the second dose of either CoronaVac (n\u0026thinsp;=\u0026thinsp;48) or ChAdOx-1 (n\u0026thinsp;=\u0026thinsp;18) to measure IgG levels for SARS-CoV-2, cytokines, and monocyte subtypes (classical, non-classical, and intermediate).\u003c/p\u003e\u003cp\u003e\u003cb\u003eResults\u003c/b\u003e\u003c/p\u003e\u003cp\u003eBoth vaccines increased specific IgG levels post-vaccination. The ChAdOx-1 group exhibited higher IgG levels, more classical, non-classical, and intermediate monocytes, and higher IL-6/IL-10 and TNF-α/IL-10 ratios compared to CoronaVac. Conversely, CoronaVac led to higher IFN-γ post-vaccination. Both vaccines promoted distinct modulation of monocyte subtypes and inflammatory status.\u003c/p\u003e\u003cp\u003e\u003cb\u003eConclusion\u003c/b\u003e\u003c/p\u003e\u003cp\u003eBoth vaccines elicited a strong antibody response, but ChAdOx-1 demonstrated better results. Distinct changes in monocyte subtypes and inflammation were observed between the two vaccine groups.\u003c/p\u003e","manuscriptTitle":"Immunization with CoronaVac or ChadOx-1 Elicit Distinct Modulation of Monocyte Subtypes and Systemic Inflammatory Status in Older Adults","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-15 08:29:35","doi":"10.21203/rs.3.rs-7085263/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"d64d4514-165d-468c-93fd-c6d2847590ef","owner":[],"postedDate":"July 15th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-03T15:08:39+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-15 08:29:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7085263","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7085263","identity":"rs-7085263","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}