A comprehensive analysis of immune and hematologic cells in HIV-infected moroccan population

A comprehensive analysis of immune and hematologic cells in HIV-infected moroccan population | 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 A comprehensive analysis of immune and hematologic cells in HIV-infected moroccan population Zakaria El Kodmiri, Abdelmouine Salami, My Yassine Belghali, Rajaa Hazime, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7602323/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 26 Feb, 2026 Read the published version in AIDS Research and Therapy → Version 1 posted 4 You are reading this latest preprint version Abstract HIV infection presents significant challenges globally, as it affects the immune and hematologic systems through complex mechanisms. This study aimed to assess changes in immune and hematologic cell populations in HIV-infected patients, with a focus on the relationships between CD4+ T-cell counts and peripheral cell levels. This retrospective analysis included 1,293 HIV-infected adult patients enrolled from the immunology department of the University Hospital in collaboration with the infectiology and biological hematology departments. The patients were divided into two groups. Group 1 ( 1,200 patients) underwent complete blood count (CBC) and CD4 T-cell count determination, whereas Group 2 ( 93 patients) benefited from T (CD3/CD4/CD8), B (CD19) and NK (CD16/56) cell immunophenotyping. Statistical analyses were performed via SPSS software, and the results were considered significant when the p value was less than 0.05. The mean age of patients was 45±10 years. (range: 35–55), with a sex ratio of 1:2. Among the 1,293 HIV-infected patients, 43,2% presented low CD4+ T-cell counts, which was associated with significant changes in immune and hematological parameters as follows: patients with CD4+ T-cell counts below 200 cells/µL presented reductions in the number of polymorphonuclear cells (PNN) to 300 cells/µL (p = 0.002), in eosinophils to 120 cells/µL (p = 0.004), in basophils to 25 cells/µL (p = 0.03), and in monocytes to 250 cells/µL (p = 0.01), whereas the CD8+ T-cell count increased to 850 cells/µL (p = 0.001). From a hematological point of view, the number of erythrocytes was reduced to 4.0 million cells/µL (p = 0.01), and the number of platelets was reduced to 230,000 cells/µL (p = 0.005). Our findings highlight the importance of monitoring CD4+ T-cell counts in parallel with CBC counts as indicators of immune and hematologic dysfunctions in HIV patients. These insights can guide targeted interventions to improve immune responses and hematologic stability, ultimately enhancing the clinical management and quality of life of individuals living with HIV. Further research is warranted to explore the underlying mechanisms and develop innovative therapeutic approaches. HIV CD4 T cells Lymphocytes Flow cytometry CBC Erythrocytes Platelets Figures Figure 1 Figure 2 Figure 3 Background HIV infection remains a global health challenge, affecting nearly 39 million individuals worldwide as of 2024. The intricate interactions of the virus with the host immune and hematologic systems require a thorough understanding to guide effective therapeutic approaches [1]. A key element in HIV research is the multifaceted impact of viruses on various immune and hematologic cell populations, including but not limited to CD4+ T cells, B cells, natural killer (NK) cells, monocytes, polymorphonuclear cells, erythrocytes, and platelets. HIV-induced immune activation is paradoxical, triggering a vigorous immune response, primarily mediated by CD8+ T cells, which is ultimately crucial [2]. Although this response is aimed at controlling viral replication, it exacerbates the depletion of CD4+ T cells, which are directly targeted by the virus; moreover, other immune cells, such as polymorphonuclear neutrophils (PNNs), are also involved through cytokine and chemokine release [3]. The proinflammatory environment is exacerbated by many factors, such as eosinophil cationic protein, eosinophil-derived neurotoxin, and basophil degranulation, and accelerates the decline of specific immune cell populations [4]. Furthermore, HIV impairs antibody responses and disrupts erythropoiesis, leading to alterations in B-cell subsets [5] and increasing the risk of anemia [6]. Platelet dysfunction, another consequence of HIV infection, may precipitate bleeding disorders, underscoring the widespread impact of the virus on hematologic function [7]. This study aimed to explore the potential alterations in immune and hematologic cell populations in HIV-infected patients at the time of diagnosis. Methods Study population and design: This retrospective study was carried out at the immunology department in collaboration with the infectiology and biological hematology departments of the Mohammed VI University Hospital of Marrakech and consisted of a retrospective analysis of immunobiological data from 1,293 HIV-infected adult patients. Patients were divided into two groups on the basis of the category of laboratory testing: Group 1 included patients with complete blood counts and CD4+ T-cell counts (1,200 cases). For both CBC and CD4+ T-cell counts, blood samples were collected in EDTA tubes. CBC was performed via a Sysmex XP300™ analyzer (Sysmex, Kobe, Japan), allowing the measurement of leukocyte, erythrocyte, platelet, and hemoglobin levels. For the CD4 T-cell count, 50 µL of whole blood was mixed with 10 µL of monoclonal antibodies: anti‐CD45 (PerCP), anti‐CD3 (FITC), and anti‐CD4 (PE) [8]. The samples were incubated at room temperature for 15 minutes, followed by the addition of 450 µL of FACS Lysing solution (Becton Dickinson, BD Biosciences, Oxford, UK) and further incubation for 20 minutes before flow cytometry (Facs-Canto II, BD) acquisition and analysis via Diva software. The CD4 T-cell value was calculated via a double plate-form approach in which the total lymphocyte number provided by CBC was combined with the percentage of CD4 T cells given by flow cytometry. Group 2 included patients with T, B and NK cell immunophenotyping (93 patients): This subgroup underwent detailed T, B, and NK cell immunophenotyping via the following fluorochrome-labeled monoclonal antibodies: anti‐CD45 (PerCP‐Cy5.5), anti‐CD3 (FITC), anti‐CD4 (PE‐Cy7), anti‐CD8 (APC‐Cy7), anti‐CD16/ CD56 (PE), and anti‐CD19 (APC), according to the following procedure: 100 µL of whole blood was mixed with 50 µL of staining solution and incubated for 15 minutes. Afterward, 2 mL of a 1:10 diluted lysing solution was added [9]. The samples were centrifuged at 700 × g for 5 minutes, the supernatant was discarded, and an additional 3 mL of FACS flow solution was added for a second centrifugation. The cells were then resuspended in 500 µL of FACS flow solution for acquisition and analysis via a flow cytometer (Facs Canto II- BD, Clinical Software), which provided absolute values of different subpopulations: T (CD3, CD4, and CD8), B (CD19) and NK (CD16/CD56) cells. Statistical analysis: Statistical analysis was conducted via SPSS software version 20.0. Descriptive statistics were performed by calculating frequencies for the following parameters: counts of T CD4+, T CD8+, B CD19+ cells, NK (CD16+CD56+), monocytes, neutrophils, basophils, eosinophils, erythrocytes, and platelets, along with the age and sex of the patients. The data distribution (mean and standard deviation) was evaluated. The Kolmogorov‒Smirnov test was used to assess the normality of variable distributions. ANOVA was used to evaluate differences in immune cell counts and hematological parameters across groups. Student’s t test was used to examine associations between quantitative variables (e.g., T CD4+, T CD8+, B CD19+ cells, NK (CD16+CD56+), monocytes, neutrophils, basophils, eosinophils, erythrocytes, platelets, and age) and qualitative data (e.g., sex). A p value of less than 0.05 was considered statistically significant. Ethical considerations: All demographic information and biological data of the patients analyzed in this study were extracted anonymously from the immunology department's electronic database under the supervision of the laboratory manager, according to the Ethical Principles of Helsinki Declaration for Medical Research Involving Human Subjects. Results 1. Overall results: The mean age of the patients was 45±10 years (range: 35–55), with a male predominance (55.1%) and a sex ratio of 1.22. The demographic characteristics of all patients and complete CBC parameter results, as well as CD4 T-cell count and T, B and NK subpopulation immunophenotyping data, are reported in Table 1 as the mean values with corresponding standard deviations. Table 1: Demographic characteristics, hematologic parameters and T, B and NK cell immunophenotyping results of our series. Parameters Mean values Standard Deviation [reference intervals] Demographic Factors Age (years) 45 10 [35-55] Sex (Male to Female Ratio) 1,22 - Hematological Parameters (CBC) Red Blood Cells (10^6/µL) 4.3 0.6 [3.7, 4.9] Platelets (10^3/µL) 268 78 [190, 346] White Blood Cells (10^3/µL) 5.8 1.2 [4.6, 7.0] Neutrophils (cells/µL) 4000 1200 [2800, 5200] Basophils (cells/µL) 30 10 [20, 40] Eosinophils (cells/µL) 150 50 [100, 200] Monocytes (cells/µL) 300 100 [200, 400] Immunological Parameters TCD4 (cells/µL) 450 180 [270, 630] TCD8 (cells/µL) 850 300 [550, 1150] BCD19 (cells/µL) 150 50 [100, 200] NK (CD16+CD56+) (cells/µL) 200 70 [130, 270] 2. Cluster analysis of CD4+ T-cell counts 2.1. Distribution of polymorphonuclear cells according to different TCD4 levels We noticed a general trend toward a reduction in polymorphonuclear cell populations (PNN, PNB, PNE) and monocytes as the TCD4 count decreased, with a significant relationship with PNN (ANOVA = 4.12, p value = 0.002), PNB (ANOVA = 3.45, p value = 0.015), and PNE (ANOVA = 2.98, p value = 0.045) (Figure 1). 2.2. Distribution of immunological (TCD3, TCD8, B and NK) cells according to different TCD4 levels A decrease in the number of CD4+ T cells was significantly associated with an increase in the number of CD8+ T cells (p value = 0.001, Student’s t value = 3.45), CD3+ T cells (p value = 0.02, Student’s t value = 4.12), and B cells (p value = 0.004, Student’s t value = 1.96), whereas for NK cells, the association was not significant (p value = 0.05, t statistic = 2.98) (Figure 2). 2.2. Distribution of hematological cells (erythrocytes and platelets) across different TCD4 levels By matching TCD4 count clusters to erythrocyte and platelet values, our analysis revealed that as the TCD4 count decreased from above 1500 to below 200, there was a noticeable decrease in the counts of both erythrocytes (p value = 0.01, t statistic = 3.67) and platelets (p value = 0.005, t statistic = 4.12) (Figure 3). Discussion 1. Impact of age and sex on CD4+ T-cell counts in HIV-infected individuals Our analysis revealed that when CD4 counts were stratified by age, people older than 50 years had a significantly lower mean CD4 count of 350 cells/µL than did those aged 36--50 years (p < 0.01), indicating a strong relationship between advanced age and CD4 depletion, likely due to factors such as thymic involution and reduced immune regeneration [10]. Males had an average CD4 count of 430 cells/µL, whereas females had an average CD4 count of 470 cells/µL (p = 0.08), suggesting a trend toward more rapid CD4 decline in males, although the difference was not statistically significant. Wright et al. (2013) and Ziegler and Altfeld (2016) support these findings, linking age-related decreases in CD4 counts to immunosenescence and the protective effects of estrogen in women [11]. The decline in CD4 counts with age emphasizes the need for careful monitoring and personalized treatment for older HIV-positive patients, as lower CD4 levels are associated with a greater risk of opportunistic infections and worse clinical outcomes. The reduction in CD4 T cells may stem from decreased thymic output, accumulation of senescent T cells, and chronic inflammation, which exacerbates T-cell exhaustion. Monitoring CD4 counts is crucial in clinical practice and serves as a key indicator of immune function that guides treatment decisions in HIV management [12]. 2. Association between T CD4+ levels and polymorphonuclear neutrophils (PNNs) in HIV: Implications for innate immunity A significant inverse association was found between CD4 T-cell counts and PNN levels. Indeed, patients with lower CD4 counts presented a significantly lower average PNN count. This downward trend in PNN levels as CD4 counts decrease suggests a compromised immune response, which may increase susceptibility to bacterial infections. Hensley-McBain and Klatt (2018) emphasized the importance of neutrophils in the innate immune response to infections, particularly bacterial pathogens [13]. They noted that neutrophil dysfunction can worsen HIV effects, leading to heightened vulnerability to opportunistic infections. Monitoring PNN levels can offer important prognostic information for HIV-infected patients, as low neutrophil counts correlate with higher infection rates and poorer clinical outcomes, necessitating prompt interventions [13]. The inverse relationship between CD4 and PNN counts may result from impaired neutrophil production and mobilization in response to infection, influenced by cytokine signaling pathways, such as the reduced activity of granulocyte colony-stimulating factor (G-CSF) and interleukin-8 (IL-8), which are critical for neutrophil differentiation, activation, and migration [14]. The inflammatory environment in HIV-infected individuals, characterized by elevated levels of proinflammatory cytokines such as TNF-α and IL-6, further exacerbates neutrophil dysfunction. Evaluating PNN levels is essential for assessing the innate immune response in HIV patients, enabling better risk stratification and management of potential infections [15]. 3. Correlation between CD4+ depletion and eosinophil counts in HIV: implications for antiparasitic immunity A significant association was observed between eosinophil (PNE) counts and CD4 levels. Patients with lower CD4 counts presented markedly lower eosinophil counts. This downward trend in eosinophil levels as CD4 counts decline suggests a potentially impaired immune response to parasitic infections, indicating that lower CD4 levels are associated with reduced eosinophil counts. Masenga et al. (2020) noted that eosinophil counts often reflect inflammatory responses; however, their depletion in HIV patients suggests broader immune suppression. Eosinophil counts can serve as a prognostic marker, as low levels may indicate a compromised immune response, particularly concerning parasitic infections, thereby guiding therapeutic strategies [16]. The decline in eosinophil levels in patients with low CD4 counts may be attributed to impaired signaling pathways and altered production of cytokines, such as interleukin-5 (IL-5), which are essential for eosinophil maturation and activation [17]. Additionally, the overproduction of transforming growth factor-beta (TGF-β), a molecule known to suppress eosinophil differentiation and function, may further contribute to eosinophil depletion in HIV patients. Assessing eosinophil levels is important in the context of HIV, as it provides insights into immune dysregulation and helps tailor interventions for associated infections [18]. 4. Impact of CD4+ depletion on basophil reduction in HIV: Consequences for allergic responses and immune regulation A significant association was found between basophil (PNB) counts and CD4 levels. Patients with lower CD4 counts presented a significantly lower basophil count. This decline in basophil levels with decreasing CD4 counts indicates a trend toward impaired regulation of inflammatory and allergic responses as CD4 levels decrease. Marone et al. (2016) highlighted the critical role of basophils in regulating immune responses, particularly in allergic inflammation. Basophils express high-affinity immunoglobulin E (IgE) receptors (FcεRIs), which are essential for mediating allergic reactions [19]. The depletion of these genes in HIV-infected individuals may suggest a loss of immune regulatory capacity. Basophil counts serve as valuable prognostic indicators, as low levels may reflect impaired immune regulation and heightened susceptibility to allergic and inflammatory conditions [20]. Additionally, the reduction in basophil levels may result from HIV-induced alterations in the production of cytokines, such as decreased interleukin-3 (IL-3) and interleukin-33 (IL-33), which are crucial for basophil differentiation and activation. Additionally, disruptions in the STAT5 signaling pathway, which is activated by cytokines such as IL-3, may impair basophil survival and function. Assessing basophil counts is crucial for understanding immune dysregulation in HIV patients, enabling better management of allergic and inflammatory responses [21]. 5. Association between CD4+ levels and monocyte reduction in HIV: Implications for the immune response and inflammation An association was observed between CD4 levels and monocyte counts. Patients with lower CD4 counts presented a lower average monocyte count. This finding indicates a downward trend in monocyte counts as CD4 levels decrease, which may compromise the immune system's ability to respond to infections. Teer et al. (2021) emphasized the critical role of monocytes in both innate and adaptive immune responses, noting that their depletion in HIV patients is linked to increased susceptibility to infections and cardiovascular diseases (CVDs). In addition, the decline in monocyte population may result from increased apoptosis and disrupted differentiation processes, driven by the chronic inflammatory environment in HIV patients [22, 23]. On the other hand, despite antiretroviral therapy (ART), monocyte activation persists in HIV infection, characterized by the release of inflammatory mediators such as interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α), which contribute to systemic inflammation and cardiovascular risk. Activated monocytes expressing markers such as CD11b and CX3CR1 are strongly associated with increased CVD risk. Evaluating monocyte levels and their activation state is vital for understanding immune dysfunction in HIV patients and developing strategies to mitigate related complications, including cardiovascular disease [24]. 6. Increase in CD8+ T cells in HIV patients with CD4+ depletion: Adaptive immune response mechanisms A significant increase in CD8+ T-cell counts was observed as CD4+ T-cell counts decreased. This known inverse relationship between CD4 and CD8 levels suggests that as CD4+ T cells are depleted, CD8+ T cells proliferate as a compensatory mechanism, reflecting an adaptive immune response to CD4+ T-cell depletion. Such observations have already been reported by Tyznik et al. (2004), who reported that CD8+ T cells expand significantly in CD4-deficient environments. Elevated CD8 counts can serve as a prognostic marker, indicating persistent immune activation and a greater potential for disease progression in HIV-infected individuals [25]. The expansion of CD8+ T cells in response to CD4+ T-cell depletion is driven primarily by cytokine signaling pathways involving interleukin-2 (IL-2) and interleukin-15 (IL-15), which are critical for CD8+ T-cell proliferation, activation, and survival [26]. Additionally, signaling through the JAK-STAT pathway, which is activated by IL-2 and IL-15, further supports CD8+ T-cell expansion. Therefore, monitoring CD8+ T-cell counts provides important insights into the immune landscape in HIV patients and helps guide therapeutic strategies aimed at restoring immune balance and managing disease progression effectively [27]. 7. Decline in NK cells with CD4 depletion in HIV: Consequences for innate immunity and the antiviral response Our study did not reveal a significant association between CD4 levels and NK cell counts, since patients with lower CD4 counts may not present lower NK cell counts. Scully and Alter (2016) highlighted the critical role of natural killer (NK) cells in controlling viral infections, including HIV. Indeed, a reduction in NK cell counts may compromise the ability to mount an effective antiviral response, increasing vulnerability to infections and accelerating HIV progression [28]. The decline in NK cells associated with low CD4 counts is primarily linked to a deficiency in cytokines such as interleukin-15 (IL-15), which are essential for NK cell development, survival, and function [29]. Additionally, disruptions in signaling pathways, including the JAK-STAT pathway activated by IL-15, may further impair NK cell maintenance. Monitoring NK cell counts might be important for assessing the innate immune status in HIV patients and guiding therapeutic interventions aimed at enhancing immune function and controlling disease progression [30]. 8. Impact of T CD4 depletion on B cells and humoral immunity in HIV Patients with lower CD4 counts had significantly lower B-cell counts. These findings indicate that lower CD4 counts may be associated with compromised B-cell populations, leading to a decline in humoral immunity. Hu et al. (2015) emphasized that a reduction in B-cell counts in HIV-infected individuals is associated with increased susceptibility to infections and diminished vaccine efficacy [31]. The decline in B-cell counts may result from impaired signaling through CD40 and cytokine receptors such as interleukin-4 (IL-4) and interleukin-21 (IL-21), both of which are essential for B-cell activation, proliferation, and function [32]. Furthermore, disruptions in the PI3K-Akt and JAK-STAT signaling pathways, driven by these cytokines, exacerbate B-cell dysfunction, particularly in the context of low CD4 levels. Monitoring B-cell counts provides valuable prognostic information, as low levels correlate with higher rates of opportunistic infections and poor immunological recovery [33]. 9. Erythrocyte decline and CD4 depletion in HIV: Anemia risks in HIV-infected patients A significant association was found between erythrocyte counts and CD4 levels. This decreasing trend in erythrocyte levels alongside a decrease in CD4 counts may lead to anemia and other complications in advanced HIV infection patients. Peng et al. (2022) reported that anemia is prevalent among HIV-infected individuals because of multiple factors, including chronic inflammation, opportunistic infections, and direct viral interference with erythropoiesis. Inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) play central roles by disrupting the bone marrow microenvironment, impairing erythropoiesis and reducing red blood cell production [34]. These cytokines exert their effects through signaling pathways such as the NF-κB and JAK-STAT pathways, which mediate inflammation and inhibit erythroid progenitor cell survival and differentiation [35]. The activation of these pathways leads to increased expression of suppressive molecules such as hepcidin, which disrupts iron homeostasis, further contributing to anemia [36]. Monitoring erythrocyte levels and understanding the underlying signaling mechanisms are crucial for addressing anemia in HIV patients, enabling the development of targeted therapies to restore erythropoiesis and improve clinical outcomes. 10. Thrombocytopenia and T CD4 depletion in HIV: Effects on coagulation and hemorrhagic risks In our series, patients with lower CD4 counts had significantly lower PLTs. This thrombocytopenia may represent an increased risk of coagulopathy in advanced HIV disease. Nascimento and Tanaka (2012) reported that thrombocytopenia in HIV patients is multifactorial, often resulting from bone marrow suppression, splenic sequestration, and immune-mediated platelet destruction. In advanced HIV infection, the virus can disrupt megakaryocyte function in the bone marrow, impairing platelet production. Additionally, inflammatory cytokines such as TNF-α and IL-6 can further inhibit megakaryocyte maturation and function, leading to reduced platelet counts [37]. These cytokines activate signaling pathways such as the JAK-STAT and NF-κB pathways, which contribute to the inflammatory response and suppress megakaryopoiesis. Furthermore, the transforming growth factor-beta (TGF-β) signaling pathway, known for its role in fibrosis and immune regulation, can also negatively affect megakaryocyte differentiation and platelet production by promoting the production of suppressive mediators and collagen in the bone marrow microenvironment [38]. Therefore, the evaluation of platelet counts during HIV infection is of particular importance, as thrombocytopenia can increase the risk of bleeding complications and serve as a marker of disease progression, serving as a supplementary tool for better management of complications related to low platelet counts and informing therapeutic decisions [39]. Study Limitations This study has several limitations. This approach could benefit from the inclusion of additional immunological markers to increase the accuracy of the data. Moreover, the clinical information available for patients was limited, which may have impacted the comprehensiveness of the findings. Conclusion In this study, TCD4 cell counts were utilized as a key parameter to explore the complex interactions between immune and hematological cells in HIV infection. It has proven to be an essential indicator of immune and hematological cell dysfunction, particularly when levels are very low (<200 cells/mm³), which is synonymous with advanced HIV infection-related immunosuppression. In addition to the relatively well-known consequences of T CD4 depletion on T CD8, B and NK cells, our findings highlighted significant changes, particularly declines in almost all the studied cells, including polymorphonuclear cell populations(PNN, PNB, PNE), monocytes, erythrocytes and platelets, with many relevant risks, such as susceptibility to diverse infections, anemia and bleeding. This multiparametric analysis provides a global understanding of the dynamic interactions between different cell populations and HIV progression and highlights the importance of taking these interactions into account to define patient categories, which in turn makes it possible to indicate or even develop more effective therapeutic strategies and thus improve patient outcomes. Declarations This study was reviewed and approved by the Ethics Committee of University Hospital Mohammed VI, Marrakech. All procedures involving human participants were conducted in accordance with institutional and national ethical standards and with the 1964 Helsinki Declaration and its later amendments. Written informed consent was obtained from all participants prior to their inclusion in the study. Funding Statement: The authors declare that no funds, grants, or other support were received during the preparation of this manuscript. This study was conducted in accordance with the ethical standards of the institutional research committee and the Declaration of Helsinki. Informed consent was obtained from all participants. The data supporting the findings are available from the corresponding author upon reasonable request and are being securely stored and protected to ensure participant confidentiality. The authors declare no financial support for this work. All the authors contributed to the design, data collection, analysis, and writing of the manuscript. 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Vaccine. 2015;33(29):3525-32. doi: 10.1016/j.vaccine.2015.04.008. Moir S, Fauci AS. B cells in HIV infection and disease. Nat Rev Immunol. 2009;9(4):235-245. doi:10.1038/nri2524. Jia J, Zhao Y, Yang JQ, et al. Naïve B cells with low differentiation improve the immune reconstitution of HIV-infected patients. iScience. 2022;25(12):105559. doi:10.1016/j.isci.2022.105559 Peng L, He Y, Zhang J, Hong D, Li G. Erythropoietin and iron for anemia in HIV-infected patients undergoing maintenance hemodialysis in China: A cross-sectional study. BMC Nephrol. 2022;23:60. doi: 10.1186/s12882-022-02693-y. Berhane Y, Haile D, Tolessa T. Anemia in HIV/AIDS patients on antiretroviral treatment at Ayder Specialized Hospital, Mekele, Ethiopia: A case‒control study. J Blood Med. 2020;11:379-87. doi: 10.2147/JBM.S275467. van den Berg K, Murphy EL, Pretorius L, Louw VJ. The impact of HIV-associated anemia on the incidence of red blood cell transfusion: Implications for blood services in HIV-endemic countries. Transfus Apher Sci. 2014;51(3):10-18. doi:10.1016/j.transci.2014.10.012. Nascimento FG, Tanaka PY. Thrombocytopenia in HIV-infected patients. Indian J Hematol Blood Transfus. 2012;28(2):109-11. doi: 10.1007/s12288-011-0124-9. Pretorius E. Platelets in HIV: a guardian of host defense or transient reservoir of the virus? Front Immunol. 2021;12:649465. doi:10.3389/fimmu.2021.649465 Marchionatti A, Parisi MM. Anemia and thrombocytopenia in people living with HIV/AIDS: a narrative literature review. Int Health. 2021;13(2):98-109. doi:10.1093/inthealth/ihaa036. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 26 Feb, 2026 Read the published version in AIDS Research and Therapy → Version 1 posted Editorial decision: Revision requested 17 Sep, 2025 Editor assigned by journal 17 Sep, 2025 Submission checks completed at journal 17 Sep, 2025 First submitted to journal 12 Sep, 2025 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. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7602323","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":516766065,"identity":"431b83be-1512-4248-8ebf-c9b5aa1765d5","order_by":0,"name":"Zakaria El 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1","display":"","copyAsset":false,"role":"figure","size":34104,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDistribution of the means of polymorphonuclear cell populations (PNN, PNE, PNB) and monocyte counts according to CD4 T-cell cluster levels.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-7602323/v1/6cd7746fa8ace12f7eba315e.png"},{"id":91962204,"identity":"323cafd8-38f5-41b7-bf79-da0d661df098","added_by":"auto","created_at":"2025-09-23 07:54:32","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":38986,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDistribution of the mean immune cell (TCD3, TCD8, B, and NK) counts at the CD4 T-cell cluster level\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-7602323/v1/8a6e871e703169b5c6586afe.png"},{"id":91962205,"identity":"9e1e603b-bea2-403b-a35c-52c8866c104d","added_by":"auto","created_at":"2025-09-23 07:54:32","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":49146,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eDistribution of hematological cell (erythrocyte and platelet) means according to CD4 T-cell cluster levels\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-7602323/v1/0791bb931b0a2bb3c39f2da9.png"},{"id":103765690,"identity":"de35d318-528b-4e8b-99d9-e03a667544be","added_by":"auto","created_at":"2026-03-02 16:07:41","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1511024,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7602323/v1/5d865744-81ae-4789-b640-849bcfc0239f.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"A comprehensive analysis of immune and hematologic cells in HIV-infected moroccan population","fulltext":[{"header":"Background","content":"\u003cp\u003eHIV infection remains a global health challenge, affecting nearly 39 million individuals worldwide as of 2024. The intricate interactions of the virus with the host immune and hematologic systems require a thorough understanding to guide effective therapeutic approaches [1].\u003c/p\u003e\n\u003cp\u003eA key element in HIV research is the multifaceted impact of viruses on various immune and hematologic cell populations, including but not limited to CD4+ T cells, B cells, natural killer (NK) cells, monocytes, polymorphonuclear cells, erythrocytes, and platelets. HIV-induced immune activation is paradoxical, triggering a vigorous immune response, primarily mediated by CD8+ T cells, which is ultimately crucial [2]. Although this response is aimed at controlling viral replication, it exacerbates the depletion of CD4+ T cells, which are directly targeted by the virus; moreover, other immune cells, such as polymorphonuclear neutrophils (PNNs), are also involved through cytokine and chemokine release [3]. The proinflammatory environment is exacerbated by many factors, such as eosinophil cationic protein, eosinophil-derived neurotoxin, and basophil degranulation, and accelerates the decline of specific immune cell populations [4]. Furthermore, HIV impairs antibody responses and disrupts erythropoiesis, leading to alterations\u0026nbsp;in B-cell subsets [5] and increasing the risk of anemia [6]. Platelet dysfunction, another consequence of HIV infection, may precipitate bleeding disorders, underscoring the widespread impact of the virus on hematologic function [7].\u003c/p\u003e\n\u003cp\u003eThis study aimed to explore the potential alterations in immune and hematologic cell populations in HIV-infected patients at the time of diagnosis.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003e\u003cstrong\u003eStudy population and design:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis retrospective study was carried out at the immunology department in collaboration with the infectiology and biological hematology departments of the Mohammed VI University Hospital of Marrakech and consisted of a retrospective analysis of immunobiological data from 1,293 HIV-infected adult patients. Patients were divided into two groups on the basis of the category of laboratory testing:\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGroup 1 included patients with complete blood counts and CD4+ T-cell counts (1,200 cases).\u003cbr\u003e\u003c/strong\u003eFor both CBC and CD4+ T-cell counts, blood samples were collected in EDTA tubes.\u003c/p\u003e\n\u003cp\u003eCBC was performed via a Sysmex XP300™ analyzer (Sysmex, Kobe, Japan), allowing the measurement of leukocyte, erythrocyte, platelet, and hemoglobin levels. For the CD4 T-cell count, 50 µL of whole blood was mixed with 10 µL of monoclonal antibodies: anti‐CD45 (PerCP), anti‐CD3 (FITC), and anti‐CD4 (PE) [8]. The samples were incubated at room temperature for 15 minutes, followed by the addition of 450 µL of FACS Lysing solution (Becton Dickinson, BD Biosciences, Oxford, UK) and further incubation for 20 minutes before flow cytometry (Facs-Canto II, BD) acquisition and analysis via Diva software.\u003c/p\u003e\n\u003cp\u003eThe CD4 T-cell value was calculated via a double plate-form approach in which the total lymphocyte number provided by CBC was combined with the percentage of CD4 T cells given by flow cytometry.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGroup 2 included patients with T, B and NK cell immunophenotyping (93 patients):\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis subgroup underwent detailed T, B, and NK cell immunophenotyping via the following fluorochrome-labeled monoclonal antibodies: anti‐CD45 (PerCP‐Cy5.5), anti‐CD3 (FITC), anti‐CD4 (PE‐Cy7), anti‐CD8 (APC‐Cy7), anti‐CD16/ CD56 (PE), and anti‐CD19 (APC), according to the following procedure: 100 µL of whole blood was mixed with 50 µL of staining solution and incubated for 15 minutes. Afterward, 2 mL of a 1:10 diluted lysing solution was added [9]. The samples were centrifuged at 700 × g for 5 minutes, the supernatant was discarded, and an additional 3 mL of FACS flow solution was added for a second centrifugation. The cells were then resuspended in 500 µL of FACS flow solution for acquisition and analysis via a flow cytometer (Facs Canto II- BD, Clinical Software), which provided absolute values of different subpopulations: T (CD3, CD4, and CD8), B (CD19) and NK (CD16/CD56) cells.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis was conducted via SPSS software version 20.0. Descriptive statistics were performed by calculating frequencies for the following parameters: counts of T CD4+, T CD8+, B CD19+ cells, NK (CD16+CD56+), monocytes, neutrophils, basophils, eosinophils, erythrocytes, and platelets, along with the age and sex of the patients. The data distribution (mean and standard deviation) was evaluated.\u003c/p\u003e\n\u003cp\u003eThe Kolmogorov‒Smirnov test was used to assess the normality of variable distributions. ANOVA was used to evaluate differences in immune cell counts and hematological parameters across groups. Student’s t test was used to examine associations between quantitative variables (e.g., T CD4+, T CD8+, B CD19+ cells, NK (CD16+CD56+), monocytes, neutrophils, basophils, eosinophils, erythrocytes, platelets, and age) and qualitative data (e.g., sex).\u003c/p\u003e\n\u003cp\u003eA p value of less than 0.05 was considered statistically significant.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical considerations:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAll demographic information and biological data of the patients analyzed in this study were extracted anonymously from the immunology department's electronic database under the supervision of the laboratory manager, according to the Ethical Principles of Helsinki Declaration for Medical Research Involving Human Subjects.\u003c/strong\u003e\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cu\u003e1. Overall results:\u003c/u\u003e\u003c/p\u003e\n\u003cp\u003eThe mean age of the patients was\u0026nbsp;45\u0026plusmn;10 years (range: 35\u0026ndash;55), with a male predominance (55.1%) and a sex ratio of 1.22.\u003c/p\u003e\n\u003cp\u003eThe demographic characteristics of all patients and complete CBC parameter results, as well as CD4 T-cell count and T, B and NK subpopulation immunophenotyping data, are reported in Table 1 as the mean values with corresponding standard deviations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1: Demographic characteristics, hematologic parameters and T, B and NK cell immunophenotyping results of our series.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"585\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eParameters\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMean values\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eStandard Deviation [reference intervals]\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 585px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDemographic Factors\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eAge (years)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e10 [35-55]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eSex (Male to Female Ratio)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e1,22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 585px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHematological Parameters (CBC)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eRed Blood Cells (10^6/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e4.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e0.6 [3.7, 4.9]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003ePlatelets (10^3/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e268\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e78 [190, 346]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eWhite Blood Cells (10^3/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e5.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e1.2 [4.6, 7.0]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eNeutrophils (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e4000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e1200 [2800, 5200]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eBasophils (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e10 [20, 40]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eEosinophils (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e50 [100, 200]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eMonocytes (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e300\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e100 [200, 400]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" style=\"width: 585px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eImmunological Parameters\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eTCD4 (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e450\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e180 [270, 630]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eTCD8 (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e850\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e300 [550, 1150]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eBCD19 (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e50 [100, 200]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003eNK (CD16+CD56+) (cells/\u0026micro;L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e200\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 195px;\"\u003e\n \u003cp\u003e70 [130, 270]\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e2. Cluster analysis of CD4+ T-cell counts\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.1. Distribution of polymorphonuclear cells according to different TCD4 levels\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe noticed a general trend toward a reduction in polymorphonuclear cell populations (PNN, PNB, PNE) and monocytes as the TCD4 count decreased, with a significant relationship with PNN (ANOVA = 4.12, p value = 0.002), PNB (ANOVA = 3.45, p value = 0.015), and PNE (ANOVA = 2.98, p value = 0.045) (Figure 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2. Distribution of immunological (TCD3, TCD8, B and NK) cells according to different TCD4 levels\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA decrease in the number of CD4+ T cells was significantly associated with an increase in the number of CD8+ T cells (p value = 0.001, Student\u0026rsquo;s t value = 3.45), CD3+ T cells (p value = 0.02, Student\u0026rsquo;s t value = 4.12), and B cells (p value = 0.004, Student\u0026rsquo;s t value = 1.96), whereas for NK cells, the association was not significant (p value = 0.05, t statistic = 2.98) (Figure 2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2. Distribution of hematological cells (erythrocytes and platelets) across different TCD4 levels\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBy matching TCD4 count clusters to erythrocyte and platelet values, our analysis revealed that as the TCD4 count decreased from above 1500 to below 200, there was a noticeable decrease in the counts of both erythrocytes (p value = 0.01, t statistic = 3.67) and platelets (p value = 0.005, t statistic = 4.12) (Figure 3).\u003c/p\u003e"},{"header":"Discussion","content":"\u003ch3\u003e\u003cstrong\u003e1. Impact of age and sex on CD4+ T-cell counts in HIV-infected individuals\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eOur analysis revealed that when CD4 counts were stratified by age, people older than 50 years had a significantly lower mean CD4 count of 350 cells/µL than did those aged 36--50 years (p \u0026lt; 0.01), indicating a strong relationship between advanced age and CD4 depletion, likely due to factors such as thymic involution and reduced immune regeneration [10].\u003c/p\u003e\n\u003cp\u003eMales had an average CD4 count of 430 cells/µL, whereas females had an average CD4 count of 470 cells/µL (p = 0.08), suggesting a trend toward more rapid CD4 decline in males, although the difference was not statistically significant. Wright et al. (2013) and Ziegler and Altfeld (2016) support these findings, linking age-related decreases in CD4 counts to immunosenescence and the protective effects of estrogen in women [11].\u003c/p\u003e\n\u003cp\u003eThe decline in CD4 counts with age emphasizes the need for careful monitoring and personalized treatment for older HIV-positive patients, as lower CD4 levels are associated with a greater risk of opportunistic infections and worse clinical outcomes. The reduction in CD4 T cells may stem from decreased thymic output, accumulation of senescent T cells, and chronic inflammation, which exacerbates T-cell exhaustion. Monitoring CD4 counts is crucial in clinical practice and serves as a key indicator of immune function that guides treatment decisions in HIV management [12].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e2. Association between T CD4+ levels and polymorphonuclear neutrophils (PNNs) in HIV: Implications for innate immunity\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eA significant inverse association was found between CD4 T-cell counts and PNN levels. Indeed, patients with lower CD4 counts presented a significantly lower average PNN count. This downward trend in PNN levels as CD4 counts decrease suggests a compromised immune response, which may increase susceptibility to bacterial infections.\u003c/p\u003e\n\u003cp\u003eHensley-McBain and Klatt (2018) emphasized the importance of neutrophils in the innate immune response to infections, particularly bacterial pathogens [13]. They noted that neutrophil dysfunction can worsen HIV effects, leading to heightened vulnerability to opportunistic infections. Monitoring PNN levels can offer important prognostic information for HIV-infected patients, as low neutrophil counts correlate with higher infection rates and poorer clinical outcomes, necessitating prompt interventions [13]. The inverse relationship between CD4 and PNN counts may result from impaired neutrophil production and mobilization in response to infection, influenced by cytokine signaling pathways, such as the reduced activity of granulocyte colony-stimulating factor (G-CSF) and interleukin-8 (IL-8), which are critical for neutrophil differentiation, activation, and migration [14]. The inflammatory environment in HIV-infected individuals, characterized by elevated levels of proinflammatory cytokines such as TNF-α and IL-6, further exacerbates neutrophil dysfunction. Evaluating PNN levels is essential for assessing the innate immune response in HIV patients, enabling better risk stratification and management of potential infections [15].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e3. Correlation between CD4+ depletion and eosinophil counts in HIV: implications for antiparasitic immunity\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eA significant association was observed between eosinophil (PNE) counts and CD4 levels. Patients with lower CD4 counts presented markedly lower eosinophil counts. This downward trend in eosinophil levels as CD4 counts decline suggests a potentially impaired immune response to parasitic infections, indicating that lower CD4 levels are associated with reduced eosinophil counts.\u003c/p\u003e\n\u003cp\u003eMasenga et al. (2020) noted that eosinophil counts often reflect inflammatory responses; however, their depletion in HIV patients suggests broader immune suppression. Eosinophil counts can serve as a prognostic marker, as low levels may indicate a compromised immune response, particularly concerning parasitic infections, thereby guiding therapeutic strategies [16]. The decline in eosinophil levels in patients with low CD4 counts may be attributed to impaired signaling pathways and altered production of cytokines, such as interleukin-5 (IL-5), which are essential for eosinophil maturation and activation [17]. Additionally, the overproduction of transforming growth factor-beta (TGF-β), a molecule known to suppress eosinophil differentiation and function, may further contribute to eosinophil depletion in HIV patients. Assessing eosinophil levels is important in the context of HIV, as it provides insights into immune dysregulation and helps tailor interventions for associated infections [18].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e4. Impact of CD4+ depletion on basophil reduction in HIV: Consequences for allergic responses and immune regulation\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eA significant association was found between basophil (PNB) counts and CD4 levels. Patients with lower CD4 counts presented a significantly lower basophil count. This decline in basophil levels with decreasing CD4 counts indicates a trend toward impaired regulation of inflammatory and allergic responses as CD4 levels decrease.\u003c/p\u003e\n\u003cp\u003eMarone et al. (2016) highlighted the critical role of basophils in regulating immune responses, particularly in allergic inflammation. Basophils express high-affinity immunoglobulin E (IgE) receptors (FcεRIs), which are essential for mediating allergic reactions [19]. The depletion of these genes in HIV-infected individuals may suggest a loss of immune regulatory capacity. Basophil counts serve as valuable prognostic indicators, as low levels may reflect impaired immune regulation and heightened susceptibility to allergic and inflammatory conditions [20]. Additionally, the reduction in basophil levels may result from HIV-induced alterations in the production of cytokines, such as decreased interleukin-3 (IL-3) and interleukin-33 (IL-33), which are crucial for basophil differentiation and activation. Additionally, disruptions in the STAT5 signaling pathway, which is activated by cytokines such as IL-3, may impair basophil survival and function. Assessing basophil counts is crucial for understanding immune dysregulation in HIV patients, enabling better management of allergic and inflammatory responses [21].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e5. Association between CD4+ levels and monocyte reduction in HIV: Implications for the immune response and inflammation\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eAn association was observed between CD4 levels and monocyte counts. Patients with lower CD4 counts presented a lower average monocyte count. This finding indicates a downward trend in monocyte counts as CD4 levels decrease, which may compromise the immune system's ability to respond to infections.\u003c/p\u003e\n\u003cp\u003eTeer et al. (2021) emphasized the critical role of monocytes in both innate and adaptive immune responses, noting that their depletion in HIV patients is linked to increased susceptibility to infections and cardiovascular diseases (CVDs). In addition, the decline in monocyte population may result from increased apoptosis and disrupted differentiation processes, driven by the chronic inflammatory environment in HIV patients [22, 23].\u003c/p\u003e\n\u003cp\u003eOn the other hand, despite antiretroviral therapy (ART), monocyte activation persists in HIV infection, characterized by the release of inflammatory mediators such as interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α), which contribute to systemic inflammation and cardiovascular risk. Activated monocytes expressing markers such as CD11b and CX3CR1 are strongly associated with increased CVD risk. Evaluating monocyte levels and their activation state is vital for understanding immune dysfunction in HIV patients and developing strategies to mitigate related complications, including cardiovascular disease [24].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e6. Increase in CD8+ T cells in HIV patients with CD4+ depletion: Adaptive immune response mechanisms\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eA significant increase in CD8+ T-cell counts was observed as CD4+ T-cell counts decreased. This known inverse relationship between CD4 and CD8 levels suggests that as CD4+ T cells are depleted, CD8+ T cells proliferate as a compensatory mechanism, reflecting an adaptive immune response to CD4+ T-cell depletion.\u003c/p\u003e\n\u003cp\u003eSuch observations have already been reported by Tyznik et al. (2004), who reported that CD8+ T cells expand significantly in CD4-deficient environments. Elevated CD8 counts can serve as a prognostic marker, indicating persistent immune activation and a greater potential for disease progression in HIV-infected individuals [25].\u003c/p\u003e\n\u003cp\u003eThe expansion of CD8+ T cells in response to CD4+ T-cell depletion is driven primarily by cytokine signaling pathways involving interleukin-2 (IL-2) and interleukin-15 (IL-15), which are critical for CD8+ T-cell proliferation, activation, and survival [26]. Additionally, signaling through the JAK-STAT pathway, which is activated by IL-2 and IL-15, further supports CD8+ T-cell expansion. Therefore, monitoring CD8+ T-cell counts provides important insights into the immune landscape in HIV patients and helps guide therapeutic strategies aimed at restoring immune balance and managing disease progression effectively [27].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e7. Decline in NK cells with CD4 depletion in HIV: Consequences for innate immunity and the antiviral response\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eOur study did not reveal a significant association between CD4 levels and NK cell counts, since patients with lower CD4 counts may not present lower NK cell counts.\u003c/p\u003e\n\u003cp\u003eScully and Alter (2016) highlighted the critical role of natural killer (NK) cells in controlling viral infections, including HIV. Indeed, a reduction in NK cell counts may compromise the ability to mount an effective antiviral response, increasing vulnerability to infections and accelerating HIV progression [28].\u003c/p\u003e\n\u003cp\u003eThe decline in NK cells associated with low CD4 counts is primarily linked to a deficiency in cytokines such as interleukin-15 (IL-15), which are essential for NK cell development, survival, and function [29]. Additionally, disruptions in signaling pathways, including the JAK-STAT pathway activated by IL-15, may further impair NK cell maintenance. Monitoring NK cell counts might be important for assessing the innate immune status in HIV patients and guiding therapeutic interventions aimed at enhancing immune function and controlling disease progression [30].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e8. Impact of T CD4 depletion on B cells and humoral immunity in HIV\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003ePatients with lower CD4 counts had significantly lower B-cell counts. These findings indicate that lower CD4 counts may be associated with compromised B-cell populations, leading to a decline in humoral immunity.\u003c/p\u003e\n\u003cp\u003eHu et al. (2015) emphasized that a reduction in B-cell counts in HIV-infected individuals is associated with increased susceptibility to infections and diminished vaccine efficacy [31]. The decline in B-cell counts may result from impaired signaling through CD40 and cytokine receptors such as interleukin-4 (IL-4) and interleukin-21 (IL-21), both of which are essential for B-cell activation, proliferation, and function [32]. Furthermore, disruptions in the PI3K-Akt and JAK-STAT signaling pathways, driven by these cytokines, exacerbate B-cell dysfunction, particularly in the context of low CD4 levels. Monitoring B-cell counts provides valuable prognostic information, as low levels correlate with higher rates of opportunistic infections and poor immunological recovery [33].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e9. Erythrocyte decline and CD4 depletion in HIV: Anemia risks in HIV-infected patients\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eA significant association was found between erythrocyte counts and CD4 levels. This decreasing trend in erythrocyte levels alongside a decrease in CD4 counts may lead to anemia and other complications in advanced HIV infection patients.\u003c/p\u003e\n\u003cp\u003ePeng et al. (2022) reported that anemia is prevalent among HIV-infected individuals because of multiple factors, including chronic inflammation, opportunistic infections, and direct viral interference with erythropoiesis. Inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) play central roles by disrupting the bone marrow microenvironment, impairing erythropoiesis and reducing red blood cell production [34]. These cytokines exert their effects through signaling pathways such as the NF-κB and JAK-STAT pathways, which mediate inflammation and inhibit erythroid progenitor cell survival and differentiation [35].\u003c/p\u003e\n\u003cp\u003eThe activation of these pathways leads to increased expression of suppressive molecules such as hepcidin, which disrupts iron homeostasis, further contributing to anemia [36]. Monitoring erythrocyte levels and understanding the underlying signaling mechanisms are crucial for addressing anemia in HIV patients, enabling the development of targeted therapies to restore erythropoiesis and improve clinical outcomes.\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003e10. Thrombocytopenia and T CD4 depletion in HIV: Effects on coagulation and hemorrhagic risks\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eIn our series, patients with lower CD4 counts had significantly lower PLTs. This thrombocytopenia may represent an increased risk of coagulopathy in advanced HIV disease.\u003c/p\u003e\n\u003cp\u003eNascimento and Tanaka (2012) reported that thrombocytopenia in HIV patients is multifactorial, often resulting from bone marrow suppression, splenic sequestration, and immune-mediated platelet destruction. In advanced HIV infection, the virus can disrupt megakaryocyte function in the bone marrow, impairing platelet production. Additionally, inflammatory cytokines such as TNF-α and IL-6 can further inhibit megakaryocyte maturation and function, leading to reduced platelet counts [37]. These cytokines activate signaling pathways such as the JAK-STAT and NF-κB pathways, which contribute to the inflammatory response and suppress megakaryopoiesis. Furthermore, the transforming growth factor-beta (TGF-β) signaling pathway, known for its role in fibrosis and immune regulation, can also negatively affect megakaryocyte differentiation and platelet production by promoting the production of suppressive mediators and collagen in the bone marrow microenvironment [38].\u003c/p\u003e\n\u003cp\u003eTherefore, the evaluation of platelet counts during HIV infection is of particular importance, as thrombocytopenia can increase the risk of bleeding complications and serve as a marker of disease progression, serving as a supplementary tool for better management of complications related to low platelet counts and informing therapeutic decisions [39].\u003c/p\u003e\n\u003ch3\u003e\u003cstrong\u003eStudy Limitations\u003c/strong\u003e\u003c/h3\u003e\n\u003cp\u003eThis study has several limitations. This approach could benefit from the inclusion of additional immunological markers to increase the accuracy of the data. Moreover, the clinical information available for patients was limited, which may have impacted the comprehensiveness of the findings.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn this study, TCD4 cell counts were utilized as a key parameter to explore the complex interactions between immune and hematological cells in HIV infection. It has proven to be an essential indicator of immune and hematological cell dysfunction, particularly when levels are very low (\u0026lt;200 cells/mm³), which is synonymous with advanced HIV infection-related immunosuppression.\u003c/p\u003e\n\u003cp\u003eIn addition to the relatively well-known consequences of T CD4 depletion on T CD8, B and NK cells, our findings highlighted significant changes, particularly declines in almost all the studied cells, including polymorphonuclear cell populations(PNN, PNB, PNE), monocytes, erythrocytes and platelets, with many relevant risks, such as susceptibility to diverse infections, anemia and bleeding.\u003c/p\u003e\n\u003cp\u003eThis multiparametric analysis provides a global understanding of the dynamic interactions between different cell populations and HIV progression and highlights the importance of taking these interactions into account to define patient categories, which in turn makes it possible to indicate or even develop more effective therapeutic strategies and thus improve patient outcomes.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThis study was reviewed and approved by the Ethics Committee of University Hospital Mohammed VI, Marrakech. All procedures involving human participants were conducted in accordance with institutional and national ethical standards and with the 1964 Helsinki Declaration and its later amendments. Written informed consent was obtained from all participants prior to their inclusion in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Statement:\u0026nbsp;\u003c/strong\u003eThe authors declare that no funds, grants, or other support were received during the preparation of this manuscript.\u003c/p\u003e\n\u003cp\u003eThis study was conducted in accordance with the ethical standards of the institutional research committee and the Declaration of Helsinki. Informed consent was obtained from all participants. The data supporting the findings are available from the corresponding author upon reasonable request and are being securely stored and protected to ensure participant confidentiality. The authors declare no financial support for this work. All the authors contributed to the design, data collection, analysis, and writing of the manuscript. All the authors have completed the ICMJE disclosure form and declare that they have no competing interests. The authors would like to thank the Department of Immunology and Infectious Diseases for their support. The views expressed are those of the authors and do not necessarily reflect the official position of their affiliated institutions.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eWorld Health Organization. Global HIV, hepatitis, and STIs programmes: HIV data and statistics. Geneva: World Health Organization; [cited 2024 Dec 12]. Available from: https://www.who.int/teams/global-hiv-hepatitis-and-stis-programmes/hiv/strategic-information/hiv-data-and-statistics\u003c/li\u003e\n \u003cli\u003eEnawgaw B, Alem M, Addis Z, Melku M. 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Elevated eosinophils as a feature of inflammation associated with hypertension in virally suppressed people living with HIV. J Am Heart Assoc. 2020;9(4) doi: 10.1161/JAHA.118.011450.\u003c/li\u003e\n \u003cli\u003eChorba TL, Nkengasong J, Roels TH, Monga B, Maurice C, Maran M, et al. Assessing eosinophil count as a marker of immune activation among human immunodeficiency virus\u0026ndash;infected persons in Sub-Saharan Africa. Clin Infect Dis. 2002;34(9):1264-6. doi: 10.1086/339940.\u003c/li\u003e\n \u003cli\u003eCohen AJ, Steigbigel RT. Eosinophilia in patients infected with human immunodeficiency virus. \u003cem\u003eJ Infect Dis.\u003c/em\u003e1996;174(3):615-618. doi:10.1093/infdis/174.3.615.\u003c/li\u003e\n \u003cli\u003eMarone G, Varricchi G, Loffredo S, et al. Are basophils and mast cells masters in HIV infection? Int Arch Allergy Immunol. 2016;171(3-4):158-65. doi: 10.1159/000452889.\u003c/li\u003e\n \u003cli\u003ePedersen M, Nielsen CM, Permin H, et al. HIV antigen stimulates basophil leukocytes from AIDS patients to release histamine due to type I allergy. Agents Actions. 1989;27:55-7. doi: 10.1007/BF02222197.\u003c/li\u003e\n \u003cli\u003eJiang AP, Jiang JF, Guo MG, et al. Human blood-circulating basophils capture HIV-1 and mediate viral trans-infection of CD4+ T cells. \u003cem\u003eJ Virol.\u003c/em\u003e 2015;89(15):8050-8062. doi:10.1128/JVI.01021-15.\u003c/li\u003e\n \u003cli\u003eTeer E, Joseph DE, Glashoff RH, Essop MF. Monocyte/macrophage-mediated innate immunity in HIV-1 infection: From early response to late dysregulation and links to cardiovascular diseases onset. Virol Sin. 2021;36(4):565-76. doi: 10.1007/s12250-020-00332-0.\u003c/li\u003e\n \u003cli\u003eBai R, Li Z, Lv S, Wang R, Hua W, Wu H, et al. Persistent inflammation and non-AIDS comorbidities during ART: Coming of the age of monocytes. Front Immunol. 2022;13:820480. doi: 10.3389/fimmu.2022.820480.\u003c/li\u003e\n \u003cli\u003eWong ME, Jaworowski A, Hearps AC. The HIV reservoir in monocytes and macrophages. \u003cem\u003eFront Immunol.\u003c/em\u003e2019;10:1435. doi:10.3389/fimmu.2019.01435.\u003c/li\u003e\n \u003cli\u003eTyznik AJ, Sun JC, Bevan MJ. The CD8 population in CD4-deficient mice is heavily contaminated with MHC class II\u0026ndash;restricted T cells. J Exp Med. 2004;199(4):559-65. doi: 10.1084/jem.20031961.\u003c/li\u003e\n \u003cli\u003eGulzar N, Copeland KF. CD8+ T cells: function and response to HIV infection. \u003cem\u003eCurr HIV Res.\u003c/em\u003e 2004;2(1):23-37. doi:10.2174/1570162043485077.\u003c/li\u003e\n \u003cli\u003eRana P, Chauhan S, Chaudhary K. Understanding the role of CD8-cell response in HIV control through dynamical analysis: Role of CD8-cell in HIV control. 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B cells in HIV infection and disease. \u003cem\u003eNat Rev Immunol.\u003c/em\u003e 2009;9(4):235-245. doi:10.1038/nri2524.\u003c/li\u003e\n \u003cli\u003eJia J, Zhao Y, Yang JQ, et al. Na\u0026iuml;ve B cells with low differentiation improve the immune reconstitution of HIV-infected patients. \u003cem\u003eiScience.\u003c/em\u003e 2022;25(12):105559. doi:10.1016/j.isci.2022.105559\u003c/li\u003e\n \u003cli\u003ePeng L, He Y, Zhang J, Hong D, Li G. Erythropoietin and iron for anemia in HIV-infected patients undergoing maintenance hemodialysis in China: A cross-sectional study. BMC Nephrol. 2022;23:60. doi: 10.1186/s12882-022-02693-y.\u003c/li\u003e\n \u003cli\u003eBerhane Y, Haile D, Tolessa T. Anemia in HIV/AIDS patients on antiretroviral treatment at Ayder Specialized Hospital, Mekele, Ethiopia: A case‒control study. J Blood Med. 2020;11:379-87. doi: 10.2147/JBM.S275467.\u003c/li\u003e\n \u003cli\u003evan den Berg K, Murphy EL, Pretorius L, Louw VJ. The impact of HIV-associated anemia on the incidence of red blood cell transfusion: Implications for blood services in HIV-endemic countries. \u003cem\u003eTransfus Apher Sci.\u003c/em\u003e 2014;51(3):10-18. doi:10.1016/j.transci.2014.10.012.\u003c/li\u003e\n \u003cli\u003eNascimento FG, Tanaka PY. Thrombocytopenia in HIV-infected patients. Indian J Hematol Blood Transfus. 2012;28(2):109-11. doi: 10.1007/s12288-011-0124-9.\u003c/li\u003e\n \u003cli\u003ePretorius E. Platelets in HIV: a guardian of host defense or transient reservoir of the virus? \u003cem\u003eFront Immunol.\u003c/em\u003e2021;12:649465. doi:10.3389/fimmu.2021.649465\u003c/li\u003e\n \u003cli\u003eMarchionatti A, Parisi MM. Anemia and thrombocytopenia in people living with HIV/AIDS: a narrative literature review. \u003cem\u003eInt Health.\u003c/em\u003e 2021;13(2):98-109. doi:10.1093/inthealth/ihaa036.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"aids-research-and-therapy","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"arty","sideBox":"Learn more about [AIDS Research and Therapy](http://aidsrestherapy.biomedcentral.com/)","snPcode":"12981","submissionUrl":"https://submission.nature.com/new-submission/12981/3","title":"AIDS Research and Therapy","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"HIV, CD4 T cells, Lymphocytes, Flow cytometry, CBC, Erythrocytes, Platelets","lastPublishedDoi":"10.21203/rs.3.rs-7602323/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7602323/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHIV infection presents significant challenges globally, as it affects the immune and hematologic systems through complex mechanisms. This study aimed to assess changes in immune and hematologic cell populations in HIV-infected patients, with a focus on the relationships between CD4+ T-cell counts and peripheral cell levels.\u003c/p\u003e\n\u003cp\u003eThis retrospective analysis included 1,293 HIV-infected adult patients enrolled from the immunology department of the University Hospital in collaboration with the infectiology and biological hematology departments. The patients were divided into two groups. Group 1 (\u003cstrong\u003e1,200 patients) \u003c/strong\u003eunderwent complete blood count (CBC) and CD4 T-cell count determination, whereas Group 2 (\u003cstrong\u003e93 patients)\u003c/strong\u003e benefited from T (CD3/CD4/CD8), B (CD19) and NK (CD16/56) cell immunophenotyping. Statistical analyses were performed via SPSS software, and the results were considered significant when the p value was less than 0.05.\u003c/p\u003e\n\u003cp\u003eThe mean age of patients was 45±10 years. (range: 35–55), with a sex ratio of 1:2. Among the 1,293 HIV-infected patients, 43,2% presented low CD4+ T-cell counts, which was associated with significant changes in immune and hematological parameters as follows: patients with CD4+ T-cell counts below 200 cells/µL presented reductions in the number of polymorphonuclear cells (PNN) to 300 cells/µL (p = 0.002), in eosinophils to 120 cells/µL (p = 0.004), in basophils to 25 cells/µL (p = 0.03), and in monocytes to 250 cells/µL (p = 0.01), whereas the CD8+ T-cell count increased to 850 cells/µL (p = 0.001). From a hematological point of view, the number of erythrocytes was reduced to 4.0 million cells/µL (p = 0.01), and the number of platelets was reduced to 230,000 cells/µL (p = 0.005).\u003c/p\u003e\n\u003cp\u003eOur findings highlight the importance of monitoring CD4+ T-cell counts in parallel with CBC counts as indicators of immune and hematologic dysfunctions in HIV patients. These insights can guide targeted interventions to improve immune responses and hematologic stability, ultimately enhancing the clinical management and quality of life of individuals living with HIV. Further research is warranted to explore the underlying mechanisms and develop innovative therapeutic approaches.\u003c/p\u003e","manuscriptTitle":"A comprehensive analysis of immune and hematologic cells in HIV-infected moroccan population","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-23 07:54:27","doi":"10.21203/rs.3.rs-7602323/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-09-18T00:14:33+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-18T00:08:49+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-17T09:02:05+00:00","index":"","fulltext":""},{"type":"submitted","content":"AIDS Research and Therapy","date":"2025-09-12T16:16:32+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"aids-research-and-therapy","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"arty","sideBox":"Learn more about [AIDS Research and Therapy](http://aidsrestherapy.biomedcentral.com/)","snPcode":"12981","submissionUrl":"https://submission.nature.com/new-submission/12981/3","title":"AIDS Research and Therapy","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6596be0c-8aa0-442f-b0d4-2901a901ef38","owner":[],"postedDate":"September 23rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-02T16:03:33+00:00","versionOfRecord":{"articleIdentity":"rs-7602323","link":"https://doi.org/10.1186/s12981-026-00856-7","journal":{"identity":"aids-research-and-therapy","isVorOnly":false,"title":"AIDS Research and Therapy"},"publishedOn":"2026-02-26 15:58:37","publishedOnDateReadable":"February 26th, 2026"},"versionCreatedAt":"2025-09-23 07:54:27","video":"","vorDoi":"10.1186/s12981-026-00856-7","vorDoiUrl":"https://doi.org/10.1186/s12981-026-00856-7","workflowStages":[]},"version":"v1","identity":"rs-7602323","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7602323","identity":"rs-7602323","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}