Skewed Memory T Cell Subsets and Dysregulated T-Follicular Helper Cells in Allergic Rhinitis

Skewed Memory T Cell Subsets and Dysregulated T-Follicular Helper Cells in Allergic Rhinitis | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL Allergy This is a preprint and has not been peer reviewed. Data may be preliminary. 26 April 2025 V1 Latest version Share on Skewed Memory T Cell Subsets and Dysregulated T-Follicular Helper Cells in Allergic Rhinitis Authors : Maryam Jafari , Eric Hjalmarsson , Eirini Paziou 0009-0009-8029-6414 , Marianne Petro , Agneta Karlsson , Ola Winquist , Susanna Kumlien Georén , and Lars Olaf Cardell 0000-0003-0538-9580 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.174566000.03531549/v1 474 views 236 downloads Contents Abstract Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Background: Allergic rhinitis (AR) is an IgE-mediated inflammatory condition characterized by Th2-skewed immune responses to allergens. Immune alterations in memory T cells, T follicular helper (Tfh) cells, and regulatory T (Treg) cells may influence baseline immune activity in AR patients even in the absence of allergen exposure, promoting sustained cytokine dysregulation, aberrant Tfh-B cell interactions, and impaired immune regulation. Aim: This study aimed to investigate the role and function of T cell subsets in AR by analyzing PBMCs from AR patients and healthy controls, focusing on immunological differences and their implications. Methods: Peripheral blood mononuclear cells from AR patients and healthy individuals were analyzed via flow cytometry to assess Th1, Th2, Th17, Tregs, and Tfh cells. Results: Off-season shifts in T cell populations were observed in AR patients compared to healthy controls. Memory T-helper (Th) cells in AR patients showed reduced frequencies of Th1/Th17 subsets and CD49d - CD27 + memory Th cells, indicating a skewed T cell profile. T follicular helper (Tfh) cells also displayed an altered distribution: AR patients had decreased Tfh1 and Tfh1/Th17 frequencies, but increased Tfh17 and Tfh2 populations, suggesting enhanced Tfh-driven B cell support. Regulatory T (Treg) cells were similarly dysregulated, with reduced frequencies of Th1/Th17-like CD49d - CD27 + Tregs and CD49d - CD27 - Tregs, reflecting impaired immune regulation. Conclusion: Off-season memory Th cell polarization, Tfh subset skewing, and Treg dysfunction in AR likely prime patients for exaggerated immune responses upon pollen exposure. These immunologic alterations provide a mechanistic link to the heightened Th2 cytokine release and IgE-mediated inflammation that trigger AR symptoms during pollen season. Skewed Memory T Cell Subsets and Dysregulated T-Follicular Helper Cells in Allergic Rhinitis Maryam Jafari 1 , Eric Hjalmarsson 1 , Eirini Paziou 1,2 , Marianne Petro 1 , Agnetha Karlsson 2 , Ola Winqvist 3 , Susanna Kumlien Georén 1 , Lars-Olaf Cardell 1,2** 1. Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden 2. Department of Otorhinolaryngology, Karolinska University Hospital, Stockholm, Sweden 3. ABC labs, Biomedicum, Stockholm Sweden Professor Lars Olaf Cardell Division of ENT Diseases Department of Clinical Science Intervention and Technology Karolinska Institutet, Stockholm, Sweden E-mail: [email protected] KEY WORDS: Allergic Rhinitis (AR), Immune Dysregulation, Th2 Skewing, T Follicular Helper (Tfh) Cells, Regulatory T (Treg) Cells, Memory T Cells, IgE Production, Immune Tolerance KEY MESSAGES: • Off-season immune dysregulation in AR patients suggests a primed allergic response before allergen exposure. • Tfh and Treg imbalances contribute to dysregulated IgE production and immune tolerance failure. • Targeting immune subsets may improve therapeutic interventions for better AR management. CAPSULE SUMMARY This study demonstrates persistent systemic immune alterations in allergic rhinitis, including imbalances in memory Th, Tfh, and Treg cells that remain evident even outside allergen exposure. These findings suggest that allergic rhinitis is characterized by ongoing immune dysregulation beyond the pollen season. AUTHORS’ CONTRIBUTIONS Conceptualization: MJ, EH, SKG, LOC. Data Curation: MJ, EH. Formal analysis: MJ, EH. Investigation: MJ, EH. Methodology: MJ, EH, SKG, LOC. Project administration: MJ, EH, SKG, LOC. Recourses: MJ, EH, EP, MP, AK. Supervision: EH, SKG, LOC. Validation: MJ, EH. Writing-original draft: MJ. Writing-review and editing: MJ, EH, EP, MP, AK, SKG, LOC. All authors have confirmed the manuscript. ABSTRACT Background: Allergic rhinitis (AR) is an IgE-mediated inflammatory condition characterized by Th2-skewed immune responses to allergens. Immune alterations in memory T cells, T follicular helper (Tfh) cells, and regulatory T (Treg) cells may influence baseline immune activity in AR patients even in the absence of allergen exposure, promoting sustained cytokine dysregulation, aberrant Tfh-B cell interactions, and impaired immune regulation. Aim: This study aimed to investigate the role and function of T cell subsets in AR by analyzing PBMCs from AR patients and healthy controls, focusing on immunological differences and their implications. Methods: Peripheral blood mononuclear cells from AR patients and healthy individuals were analyzed via flow cytometry to assess Th1, Th2, Th17, Tregs, and Tfh cells. Results: Off-season shifts in T cell populations were observed in AR patients compared to healthy controls. Memory T-helper (Th) cells in AR patients showed reduced frequencies of Th1/Th17 subsets and CD49d - CD27⁺ memory Th cells, indicating a skewed T cell profile. T follicular helper (Tfh) cells also displayed an altered distribution: AR patients had decreased Tfh1 and Tfh1/Th17 frequencies, but increased Tfh17 and Tfh2 populations, suggesting enhanced Tfh-driven B cell support. Regulatory T (Treg) cells were similarly dysregulated, with reduced frequencies of Th1/Th17-like CD49d - CD27⁺ Tregs and CD49d - CD27 - Tregs, reflecting impaired immune regulation. Conclusion: Off-season memory Th cell polarization, Tfh subset skewing, and Treg dysfunction in AR likely prime patients for exaggerated immune responses upon pollen exposure. These immunologic alterations provide a mechanistic link to the heightened Th2 cytokine release and IgE-mediated inflammation that trigger AR symptoms during pollen season. Word Count: 255 INTRODUCTION Allergic rhinitis (AR) is a prevalent immunological disorder affecting a significant portion of the global population. It is characterized by an exaggerated T-helper 2 (Th2)-mediated immune response to environmental allergens, leading to symptoms such as nasal congestion, rhinorrhea, sneezing, and itching (1). Traditionally, AR has been viewed as a localized disorder of the nasal mucosa; however, emerging research redefines it as a systemic immune condition with implications extending beyond the nasal cavity (2). This systemic involvement highlights the role of immune compartmentalization, where localized immune responses in the nasal mucosa are closely linked with systemic alterations in peripheral blood and other tissues(1). Such systemic immune dysregulation can contribute to associated comorbidities, notably asthma, underscoring the interconnected nature of mucosal immune responses across respiratory sites (1) . AR is classically driven by allergen-specific Th2 cells, which release cytokines (IL-4, IL-5, IL-13) upon allergen exposure, orchestrating IgE production and eosinophilic inflammation (3). However, accumulating evidence suggests that other T cell subsets, including Th1, Th17, T follicular helper (Tfh), and regulatory T (Treg) cells also play important roles in AR pathogenesis (3). Th1 cells (typically producing IFN-γ) and Th17 cells (producing IL-17) can modulate immune responses and disease severity, while Tregs are essential for maintaining tolerance and suppressing excessive inflammation. Dysregulation of these cell populations contributes to immune imbalance, potentially leading to chronic allergic inflammation (4). Emerging research has broadened the immunological landscape of AR to include a variety of T cell subsets such as Th17, T follicular helper (Tfh), and regulatory T (Treg) cells, each contributing uniquely to the pathology and persistence of this disease (5). Th17 cells, often linked with neutrophilic inflammation, contribute to immune dysregulation in AR (5). Tfh cells have emerged as critical players in allergic diseases, as they reside in lymphoid follicles and support B cell-mediated antibody production, including class switching to IgE (4). Elevated Tfh2 activity has been linked to AR pathogenesis through enhanced allergen-specific IgE generation. In contrast, T follicular regulatory (Tfr) cells, a subset of Tregs, normally counterbalance Tfh-driven responses (6). Regulatory T cells, pivotal in maintaining immune tolerance, are often found to be dysfunctional or insufficient in AR, leading to an unchecked allergen-specific immune response. This complex array of immune dysregulation not only facilitates the chronicity of symptoms but also presents potential targets for therapeutic intervention (7). Memory T cells play a pivotal role in AR, as they persist between allergen exposures and can rapidly respond upon re-exposure (8). Studies of nasal tissue in perennial AR have demonstrated increased infiltration of memory CD4 + T cells in allergic individuals, correlating with elevated local Th2 cytokine production (9). This suggests that allergic patients maintain a primed pool of memory T cells skewed toward allergy-promoting phenotypes(10). Circulating immune cells in AR patients may also reflect this bias even outside the pollen season (11). Reports indicate that AR patients exhibit signs of systemic T cell activation, including higher circulating T cell counts and cytokine levels (IL-6, IL-10, IFN-γ) compared to healthy individuals(12). Notably, reductions in certain Th1 and Th17 cell populations, along with decreased expression of the Th1-associated chemokine receptor CXCR3 on CD4 + T cells, suggest an imbalance favoring Th2-driven immunity and a suppressed Th1/Th17 axis in peripheral blood (10). This immune skewing likely contributes to the persistence of AR symptoms beyond direct allergen exposure (8). CD49d (integrin α4) and CD27 (TNF receptor superfamily member 7) are crucial markers for distinguishing functionally distinct T cell subsets involved in immune regulation, migration, and activation (13). CD49d facilitates T cell adhesion and migration, enabling immune cells to infiltrate inflamed tissues (13). It plays a pivotal role in Th1 and Th17 cell trafficking, and its altered expression in AR suggests an imbalance in immune cell distribution (14). In contrast, CD27 is essential for T cell survival, differentiation, and memory formation. It is highly expressed on naïve and central memory T cells but decreases as cells differentiate into effector subsets (15). Both CD49d and CD27 serve as valuable markers for understanding immune dysregulation in allergic diseases (16) . This study was designed to use flow cytometry to investigate persistent alterations in the balance of T cell subsets in patients with AR, even outside the pollen season. MATERIALS AND METHODS The samples analyzed in this study were collected from both patients with birch and/or timothy pollen-induced seasonal allergic rhinitis (AR), as well as from healthy individuals from the same cohort, during the winter/spring of 2022, prior to the natural pollen season. A total of 24 patients diagnosed with moderate to severe seasonal AR triggered by birch and/or timothy pollen were included (Table 1). Participants met the inclusion criteria of having a confirmed history of seasonal AR due to birch and/or timothy pollen, a positive skin prick test, and an allergen-specific IgE level of at least 0.35 kE/L in blood. Exclusion criteria comprised recent upper respiratory infections (within the past two weeks), recent use of local or systemic corticosteroids (within the past two months), antihistamine intake within 24 hours prior to enrollment, and a history of chronic rhinosinusitis with or without nasal polyposis. Additionally, 32 non-allergic healthy individuals were included as controls, all with no prior history of seasonal AR and a negative result in the ImmunoCAP Rapid test (Thermo Fisher Scientific, Waltham, MA, USA). Patients (N=24) Healthy controls (N=32) Age* 36 (18-57) 31 (19-61) Gender , n (%) Male 8 (33%) 14 (44%) Female 16 (67%) 18 (56%) Allergy Only birch, n (%) 1 (4%) N/A Only timothy, n (%) 10 (42%) N/A Both birch and timothy, n (%) 13 (54%) N/A *Median (range), N/A, not applicable. Ethics Statement All participants provided written informed consent prior to inclusion, and all procedures, including the handling of patient data, were conducted in compliance with the Declaration of Helsinki and approved by the Swedish Ethical Review Authority (Diary No. 2021-00325, 2021-06514-02). Flow cytometry Peripheral blood mononuclear cells (PBMCs) were isolated following standard procedures. BD Horizon™ Fixable Viability Stain 780 (BD Biosciences) was used for live/dead cell discrimination. PBMCs were then Fc-blocked and incubated in the dark for 5 minutes at room temperature (RT). Following viability staining and Fc-blocking, the cells were stained with a flow cytometry panel (Supplementary Table 1) to analyze T cell subtypes, including naïve, memory, regulatory, and effector populations. General T cells were identified using CD3 for total T cells and CD4 for helper T cells. Memory and naïve T cells were distinguished using CD45RO for memory cells, CD45RA for naïve cells, and CD45RB for further subset classification. Regulatory T cells (Tregs) were identified based on high expression of CD25 hi expression of CD127 low expression T follicular helper (Tfh) cells were characterized by CXCR5 expression, whereas CD49d and CD27 were used to delineate Tfh subsets. Th1 cells were identified using CXCR3 and CCR6 as a Th17 marker. These Th subset definitions are based on chemokine receptor expression patterns and serve as approximations of functional states. Specifically, Th1 cells were defined as CXCR3⁺CCR6 - , Th17 as CXCR3⁻CCR6⁺, and Th1/Th17 as CXCR3⁺CCR6⁺. While commonly used in flow cytometric immunophenotyping, these markers do not directly reflect cytokine production or transcription factor expression. Additional markers included CD161 for identifying Th17 and innate-like lymphocytes, and CD38 for assessing activation status. To exclude dead cells, Fixable Viability Stain 780 was used. All antibodies and fluorochromes were obtained from BD Biosciences, as specified in Supplementary Table 1. After staining, the cells were incubated in the dark for 15 minutes at RT. Following incubation, 3 mL of PBS was added, and the cells were centrifuged at 400 × g for 5 minutes. The pellet was then resuspended in 1% paraformaldehyde (PFA) solution for fixation. Samples were analyzed with flow cytometry using an LSR Fortessa X-20, and data analysis was performed using FlowJo version 10.7.1 (both from BD Biosciences). Statistical analysis Data analysis was conducted using GraphPad Prism software (Version 10, San Diego, California). Pairwise comparisons were performed using a paired Student’s t-test. If the data did not follow a normal distribution, the Wilcoxon matched-pairs signed-rank test was applied instead. Statistical significance was indicated as follows: p < 0.05 (* ), p < 0.01 ( ** ), p < 0.001 (***) and p < 0.0001 (****). RESULTS Memory T-helper subtypes in allergic rhinitis patients To investigate differences in memory T-helper (Th) cell subsets between allergic rhinitis (AR) patients and healthy controls (HC) during the pollen-free season, we analyzed the distribution of Th1, Th1/Th17, Th17, and Th2 cells (Figure 1A-D). No significant differences were observed in the frequencies of naïve CD45RA-expressing Th1, Th17, and Th2 cells in the circulation (Figure 1A, 1C, and 1D). In contrast, CD45RO-expressing Th1 and Th17 memory cells were significantly lower in blood samples from AR patients compared to HC (Figure 1B). Further analysis of CD49d and CD27 expression within memory Th1/Th17 cell subsets revealed that the frequency of CD49d - CD27⁺ memory cells was significantly lower in AR patients compared to HC (Figure 1E). Similarly, the frequency of CD49d - CD27 - memory cells was also significantly reduced in AR patients (Figure 1H). No significant differences were observed in the frequencies of CD49d⁺CD27⁺ and CD49d⁺CD27⁻ memory cell populations (Figure 1F and 1G). Figure 1. Comparison of memory Th subtypes in allergic rhinitis patients and healthy individuals before the natural pollen season. Panels (A-D) show the percentage of memory cells within the (A) Th1, (B) Th1/Th17, (C) Th17, and (D) Th2 subsets. Panels (E-H) compare the frequency of memory Th1/Th17 cells expressing specific marker combinations: (E) CD49d - CD27 + , (F) CD27 + CD49d + , (G) CD49d + CD27 - , and (H) CD49d - CD27 - . Data are shown for healthy controls (HC; n = 32) and allergic rhinitis patients (AR; n = 24) using samples collected before the natural pollen season. Statistical comparisons were made using an unpaired Student’s t-test. * = p < 0.05, **=P<0.001; ns = not significant. T-follicular helper subtypes expressing CD49d/CD27 markers The distribution of T-follicular helper (Tfh) cell subsets co-expressing CD49d and CD27 markers also revealed notable differences between AR patients and HC (Figure 2A-D). The frequency of Tfh1 cells expressing CD49d - CD27⁺ markers was significantly lower in AR patients compared to HC (Figure 2A). Similarly, Tfh1/Th17 cells expressing CD49d - CD27⁺ markers showed a significant reduction in AR patients (Figure 2B). In contrast, the frequencies of Tfh17 and Tfh2 cells expressing CD49d⁺ CD27⁺ markers were significantly higher in AR patients compared to HC (Figure 2C and 2D). Figure 2. Comparison of Tfh subtypes expressing CD49d - CD27 + and CD49d + CD27 + markers in allergic rhinitis patients and healthy individuals before the natural pollen season. Panels (A-D) display the percentage of Tfh cells co-expressing CD49d - CD27 + or CD49d + CD27 + markers within specific subsets: (A) Tfh1 (CD49d - CD27 + ), (B) Tfh1/Th17 (CD49d - CD27 + ), (C) Tfh17 (CD49d + CD27+), and (D) Tfh2 (CD49d + CD27 + ) cells. Data are shown for healthy controls (HC; n = 32) and allergic rhinitis patients (AR; n = 24). Statistical comparisons were performed using an unpaired Student’s t-test. *** = p < 0.001; **** = p < 0.0001. Regulatory T cells subtypes expressing CD49d and CD27 markers We next assessed differences in regulatory T cell (Treg) subsets co-expressing CD49d and CD27 markers in AR patients and HC before the natural pollen season (Figure 3A-D). The analysis revealed that the frequency of Treg Th1/Th17 cells expressing CD49d - CD27⁺ markers was significantly lower in AR patients compared to HC (Figure 3A). No significant difference was observed in the frequency of Treg Th1/Th17 cells expressing CD49d⁺ CD27⁺ markers (Figure 3B). Interestingly, the percentage of CD49d - CD27 - Treg Th1/Th17 cells was also significantly lower in AR patients compared to HC (Figure 3C). No significant differences were found for CD49d + CD27 - Treg Th1/Th17 cells (Figure 3D). Figure 3. Comparison of Treg subtypes expressing CD49d and CD27 markers in allergic rhinitis patients and healthy individuals before the natural pollen season . Panels (A-D) show the percentage of Treg cells expressing specific CD49d and CD27 marker combinations within subsets: (A) Treg Th1/Th17 (CD49d - CD27 + ), (B) Treg Th1/Th17 (CD49d + CD27 + ), (C) Treg Th1/Th17 (CD49d - CD27 - ), and (D) Treg Th1/Th17 (CD49d + CD27 - ). Data are presented for healthy controls (HC; n = 32) and allergic rhinitis patients (AR; n = 24). Statistical comparisons were performed using an unpaired Student’s t-test. ** = p < 0.01; *** = p < 0.001; ns = not significant. DISCUSSION In this study, we identified several key immunological differences in peripheral T cell subsets between allergic rhinitis patients and healthy controls in the absence of seasonal allergen exposure. First, AR patients had a lower frequency of memory Th1/Th17 cells. Second, within the Th1/Th17 memory subset, AR patients showed a decrease in CD27 + , while no corresponding increase in CD27 - cells was detected. Third, circulating Tfh cells in AR were skewed: Tfh1 and Tfh1/Th17 subsets were reduced, whereas Tfh17 and Tfh2 subsets were expanded. Fourth, we observed an imbalance in regulatory T cell subpopulations characterized by a loss of CD49d - CD27 + and CD49d - CD27 - Tregs in AR patients. We discuss each of these findings in turn and consider their implications in the context of existing literature on immunological mechanisms in allergic rhinitis. Our investigation into AR reveals profound systemic immune alterations in T cell subsets, affecting memory Th, Tfh, and Treg cells even outside the pollen exposure periods. These insights underscore the complexity of AR as a chronic and systemic condition marked by persistent immune dysregulation and compartmentalization. The persistence of memory Th cells and the imbalance in Tfh and Treg cells in AR patients emphasize a systemic immunological involvement that extends beyond the nasal mucosa (17). This systemic feature of AR suggests that immune responses are not isolated to the site of allergen contact but involve multiple immune compartments including peripheral blood and potentially lymph nodes, as evidenced by altered T cell dynamics (18). Our previous research highlights the significance of T cell localization in AR, illustrating how T cell responses in the nasal mucosa can reflect broader systemic immune processes(19). The results in Figure 1 demonstrate dysregulation in memory Th1/Th17 subsets. Our findings indicate a significant reduction in both CD49d - CD27⁺ and CD49d - CD27 - memory Th1/Th17 cells. The loss of CD49d - CD27⁺ cells, which are essential for maintaining the balance between effector and regulatory responses, suggests impaired immune regulation, potentially allowing for the expansion of Th2-driven inflammation. (20). As shown in Figure 2 , the altered expression of CD49d and CD27 extends to Tfh subsets, with a significant reduction in Tfh1 (CD49d - CD27⁺) and Tfh1/Th17 (CD49d - CD27⁺) cells, accompanied by an increase in Tfh2 (CD49d⁺CD27⁺) and Tfh17 (CD49d⁺CD27⁺) subsets. This shift indicates a bias toward Tfh-mediated IgE production, further reinforcing the role of Tfh cells in allergic inflammation. This shift suggests a tendency towards Tfh-mediated IgE production, reinforcing the role of Tfh cells in allergic inflammation (6). The increase of Tfh2 and Tfh17 populations aligns with previous studies highlighting their contribution to germinal center formation and allergen-specific IgE responses, a hallmark of AR pathophysiology (21). Dysregulation in memory Th1/Th17 subsets along with shifts in Tfh and Treg cells, suggesting a deviation from immune homeostasis in AR patients (22). The observed dysregulation in Th1/Th17 subsets and Tfh cells likely results from phenotype switching or migration from peripheral blood to other compartments such as lymph nodes or nasal mucosa (23) . This migration is crucial for sustaining localized immune responses during allergen exposure and may contribute to the systemic manifestations of AR even during off-seasons (24). The role of Th17 cells extends beyond simple participation in AR pathology; their involvement is crucial in mediating neutrophilic inflammation via IL-8, a potent chemokine for neutrophil migration (25). Furthermore, the increase in Th17 cells, as we have previously documented, suggests a potential mechanism for sustained inflammation through IL8-driven neutrophil recruitment (2). This highlights the dual inflammatory pathways in AR, involving both eosinophilic and neutrophilic responses, which may contribute to disease severity and chronicity (2). The results in Figure 3 showed distinct alterations in Treg subsets, with a marked reduction in both CD49d - CD27 + and CD49d - CD27 - Th1/Th17-like Treg cell populations. This suggests a possible impairment in the suppressive function of Tregs, allowing unchecked activation of inflammatory pathways(12) . Since Tregs play a pivotal role in maintaining immune tolerance, their dysfunction in AR patients may contribute to persistent allergic inflammation (12). These findings are in line with previous studies demonstrating that a loss of functional Tregs correlates with heightened allergic responses and reduced immune tolerance in AR (12). The combined use of CD49d and CD27 to define functional subsets of Tfh and Treg cells is not widely established but provides valuable insights into T cell heterogeneity and activation state. CD27 has been used as a marker of differentiation and survival in T cells (15), while CD49d (integrin α4) plays a role in cell migration and activation (26). In our study, this dual-marker strategy allowed us to resolve phenotypic distinctions within both Tfh and Treg populations that are often difficult to distinguish using conventional markers. For instance, the loss of CD49d and CD27 in Tregs may reflect a shift toward a less suppressive or more dysfunctional phenotype. Similarly, distinct expression patterns within Tfh subsets may indicate different capacities to support B cell responses. This classification strategy is supported by previous work, which demonstrated that combined assessment of CD49d and CD27 expression can reveal functional diversity among CD4⁺ T cells in chronic immune conditions (27). CONCLUSION Given the persistent immune dysregulation observed in AR, our findings underscore the need for therapeutic strategies that address these long-standing alterations. Modulating dysregulated T cell responses, particularly by enhancing regulatory T (Treg) cell functionality or restoring the balance between memory Th and Tfh cell subsets, may provide a more comprehensive approach to AR management. Targeted strategies to restore immune balance could help control chronic inflammation and prevent disease progression. Moreover, integrating insights from various studies, including our own, may facilitate the development of more precise and effective management strategies for AR. Future treatments should prioritize correcting underlying immune dysfunction rather than solely focusing on symptom relief to achieve sustained disease control. COMPETING INTERESTS The authors declare no conflict of interest. FUNDING Swedish Research Council, Stockholm county (ALF), Konsul Berghs Stiftelse, Astma och allergiförbundet, and CIMED (centrum för innovativ medicin) AVAILABILITY OF DATA AND MATERIALS The data sets used and/or analyzed in this study are available from the corresponding author upon reasonable request. ACKNOWLEDGMENTS We thank all participants for their contributions to this study. AUTHORS LIST: Maryam jafari 1 1 https://orcid.org/0009-0009-5219-4998 Eric Hjalmarsson 2 2 https://orcid.org/0009-0007-7228-6131 Eirini Paziou 3 3 https://orcid.org/0009-0009-8029-6414 Marianne Petro 4 4 https://orcid.org/0009-0005-1996-1199 Agnetha Karlsson 5 5 https://orcid.org/0009-0005-2127-2624 Ola winqvist 6 6 https://orcid.org/0000-0003-0842-4441 Susanna Kumlien Georén 7 7 https://orcid.org/0000-0001-7292-9255 Lars Olaf Cardell 8 8 https://orcid.org/0000-0003-0538-9580 REFERENCES 1. Hjalmarsson E, Petro M, Georén SK, Winqvist O, Cardell LO. Upregulated expression of Notch1/4 - JAG-1/DLL-1 detected in allergic rhinitis. Allergy Asthma Clin Immunol. 2023;19(1):41.2. Arebro J, Ekstedt S, Hjalmarsson E, Winqvist O, Kumlien Georén S, Cardell LO. A possible role for neutrophils in allergic rhinitis revealed after cellular subclassification. Sci Rep. 2017;7:43568.3. Stelmaszczyk-Emmel A, Zawadzka-Krajewska A, Kopatys A, Demkow U. Th1, Th2, Th17, and regulatory cytokines in children with different clinical forms of allergy. Adv Exp Med Biol. 2013;788:321-8.4. Hjalmarsson E, Hellkvist L, Karlsson A, Winquist O, Kumlien Georén S, Westin U, et al. A 5-Year Open-Label Follow-up of a Randomized Double-Blind Placebo-Controlled Trial of Intralymphatic Immunotherapy for Birch and Grass Allergy Reveals Long-term Beneficial Effects. J Investig Allergol Clin Immunol. 2023;33(5):362-72.5. Shamji MH, Layhadi JA, Sharif H, Penagos M, Durham SR. Immunological Responses and Biomarkers for Allergen-Specific Immunotherapy Against Inhaled Allergens. J Allergy Clin Immunol Pract. 2021;9(5):1769-78.6. Kobayashi T, Iijima K, Dent AL, Kita H. Follicular helper T cells mediate IgE antibody response to airborne allergens. J Allergy Clin Immunol. 2017;139(1):300-13.e7.7. Iinuma T, Okamoto Y, Morimoto Y, Arai T, Sakurai T, Yonekura S, et al. Pathogenicity of memory Th2 cells is linked to stage of allergic rhinitis. Allergy. 2018;73(2):479-89.8. Yao Y, Wang ZC, Wang N, Zhou PC, Chen CL, Song J, et al. Allergen immunotherapy improves defective follicular regulatory T cells in patients with allergic rhinitis. J Allergy Clin Immunol. 2019;144(1):118-28.9. Pawankar RU, Okuda M, Okubo K, Ra C. Lymphocyte subsets of the nasal mucosa in perennial allergic rhinitis. Am J Respir Crit Care Med. 1995;152(6 Pt 1):2049-58.10. Yu X, Wang M, Cao Z. Reduced CD4(+)T Cell CXCR3 Expression in Patients With Allergic Rhinitis. Front Immunol. 2020;11:581180.11. Kim AS, Doherty TA, Karta MR, Das S, Baum R, Rosenthal P, et al. Regulatory B cells and T follicular helper cells are reduced in allergic rhinitis. J Allergy Clin Immunol. 2016;138(4):1192-5.e5.12. Noval Rivas M, Chatila TA. Regulatory T cells in allergic diseases. J Allergy Clin Immunol. 2016;138(3):639-52.13. Shamji MH, Sharif H, Layhadi JA, Zhu R, Kishore U, Renz H. Diverse immune mechanisms of allergen immunotherapy for allergic rhinitis with and without asthma. J Allergy Clin Immunol. 2022;149(3):791-801.14. Rossi B, Dusi S, Angelini G, Bani A, Lopez N, Della Bianca V, et al. Alpha4 beta7 integrin controls Th17 cell trafficking in the spinal cord leptomeninges during experimental autoimmune encephalomyelitis. Front Immunol. 2023;14:1071553.15. Remedios KA, Meyer L, Zirak B, Pauli ML, Truong HA, Boda D, et al. CD27 Promotes CD4(+) Effector T Cell Survival in Response to Tissue Self-Antigen. J Immunol. 2019;203(3):639-46.16. Pizzarello CR, Jackson CM, Herman K, Seppo AE, Rebhahn J, Scherzi T, et al. A Phenotypically Distinct Human Th2 Cell Subpopulation Is Associated With Development of Allergic Disorders in Infancy. Allergy. 2025.17. Chen M, Meng Y, Shi X, Zhu C, Zhu M, Tang H, et al. Identification of ENTPD1 as a novel biomarker linking allergic rhinitis and systemic lupus erythematosus. Sci Rep. 2024;14(1):18266.18. Varricchi G, Harker J, Borriello F, Marone G, Durham SR, Shamji MH. T follicular helper (Tfh ) cells in normal immune responses and in allergic disorders. Allergy. 2016;71(8):1086-94.19. Fransson M, Adner M, Erjefält J, Jansson L, Uddman R, Cardell LO. Up-regulation of Toll-like receptors 2, 3 and 4 in allergic rhinitis. Respir Res. 2005;6(1):100.20. Wambre E, Bajzik V, DeLong JH, O’Brien K, Nguyen QA, Speake C, et al. A phenotypically and functionally distinct human T(H)2 cell subpopulation is associated with allergic disorders. Sci Transl Med. 2017;9(401).21. Gowthaman U, Chen JS, Eisenbarth SC. Regulation of IgE by T follicular helper cells. J Leukoc Biol. 2020;107(3):409-18.22. Guo LP, Yan M, Niu RB, Liu L, Yang JR, Chen RL, et al. Role of Th2, Th17 and Treg Cells and relevant cytokines in pathogenesis of allergic rhinitis. Allergy Asthma Clin Immunol. 2024;20(1):40.23. Gosset P, Tillie-Leblond I, Malaquin F, Durieu J, Wallaert B, Tonnel AB. Interleukin-8 secretion in patients with allergic rhinitis after an allergen challenge: interleukin-8 is not the main chemotactic factor present in nasal lavages. Clin Exp Allergy. 1997;27(4):379-88.24. Pawankar R, Mori S, Ozu C, Kimura S. Overview on the pathomechanisms of allergic rhinitis. Asia Pac Allergy. 2011;1(3):157-67.25. Fan X, Shu P, Wang Y, Ji N, Zhang D. Interactions between neutrophils and T-helper 17 cells. Front Immunol. 2023;14:1279837.26. Wi T, Choi Y, Kim J, Choi YS, Pipkin ME, Choi J. Efficient gene deletion of Integrin alpha 4 in primary mouse CD4 T cells using CRISPR RNA pair-mediated fragmentation. Front Immunol. 2024;15:1445341.27. Pizzarello CR, Jackson CM, Herman K, Seppo AE, Rebhahn J, Scherzi T, et al. A Phenotypically Distinct Human Th2 Cell Subpopulation Is Associated With Development of Allergic Disorders in Infancy. Allergy. 2025;80(4):949-64. Information & Authors Information Version history V1 Version 1 26 April 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Collection Allergy Keywords allergy treament basic mechanisms biomarkers ent (rhinitis flow cytometry ige immunotherapy and tolerance induction nasal polyps...) sinusitis t cells Authors Affiliations Maryam Jafari Karolinska Institutet Institutionen for klinisk vetenskap intervention och teknik View all articles by this author Eric Hjalmarsson Karolinska Institutet Institutionen for klinisk vetenskap intervention och teknik View all articles by this author Eirini Paziou 0009-0009-8029-6414 Karolinska Institutet Institutionen for klinisk vetenskap intervention och teknik View all articles by this author Marianne Petro Karolinska Institutet Institutionen for klinisk vetenskap intervention och teknik View all articles by this author Agneta Karlsson Karolinska Institutet Institutionen for klinisk vetenskap intervention och teknik View all articles by this author Ola Winquist Medicines Company Stockholm Sweden View all articles by this author Susanna Kumlien Georén Karolinska Institutet Institutionen for klinisk vetenskap intervention och teknik View all articles by this author Lars Olaf Cardell 0000-0003-0538-9580 [email protected] Karolinska Institutet Institutionen for klinisk vetenskap intervention och teknik View all articles by this author Metrics & Citations Metrics Article Usage 474 views 236 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Maryam Jafari, Eric Hjalmarsson, Eirini Paziou, et al. Skewed Memory T Cell Subsets and Dysregulated T-Follicular Helper Cells in Allergic Rhinitis. Authorea . 26 April 2025. DOI: https://doi.org/10.22541/au.174566000.03531549/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . Format Please select one from the list RIS (ProCite, Reference Manager) EndNote BibTex Medlars RefWorks Direct import Tips for downloading citations document.getElementById('citMgrHelpLink').addEventListener('click', function() { popupHelp(this.href); return false; }); $(".js__slcInclude").on("change", function(e){ if ($(this).val() == 'refworks') $('#direct').prop("checked", false); $('#direct').prop("disabled", ($(this).val() == 'refworks')); }); View Options View options PDF View PDF Figures Tables Media Share Share Share article link Copy Link Copied! Copying failed. Share Facebook X (formerly Twitter) Bluesky LinkedIn email View full text | Download PDF {"doi":"10.22541/au.174566000.03531549/v1","type":"Article"} Now Reading: Share Figures Tables Close figure viewer Back to article Figure title goes here Change zoom level Go to figure location within the article Download figure Toggle share panel Toggle share panel Share Toggle information panel Toggle information panel Go to previous graphic Go to next graphic Go to previous table Go to next table All figures All tables View all material View all material xrefBack.goTo xrefBack.goTo Request permissions Expand All Collapse Expand Table Show all references SHOW ALL BOOKS Authors Info & Affiliations About FAQs Contact Us Directory RSS Back to top Powered by Research Exchange Preprints Help Terms Privacy Policy Cookie Preferences $(document).ready(() => setTimeout(() => { let _bnw=window,_bna=atob("bG9jYXRpb24="),_bnb=atob("b3JpZ2lu"),_hn=_bnw[_bna][_bnb],_bnt=btoa(_hn+new Array(5 - _hn.length % 4).join(" ")); $.get("/resource/lodash?t="+_bnt); },4000)); (function(){function c(){var b=a.contentDocument||a.contentWindow.document;if(b){var d=b.createElement('script');d.innerHTML="window.__CF$cv$params={r:'a010821f5a6952ad',t:'MTc3OTY2OTgzMQ=='};var a=document.createElement('script');a.src='/cdn-cgi/challenge-platform/scripts/jsd/main.js';document.getElementsByTagName('head')[0].appendChild(a);";b.getElementsByTagName('head')[0].appendChild(d)}}if(document.body){var a=document.createElement('iframe');a.height=1;a.width=1;a.style.position='absolute';a.style.top=0;a.style.left=0;a.style.border='none';a.style.visibility='hidden';document.body.appendChild(a);if('loading'!==document.readyState)c();else if(window.addEventListener)document.addEventListener('DOMContentLoaded',c);else{var e=document.onreadystatechange||function(){};document.onreadystatechange=function(b){e(b);'loading'!==document.readyState&&(document.onreadystatechange=e,c())}}}})();