Representative gating strategy to identify T lymphocyte (A-N) and B lymphocyte (O-X) subsets. HC with a value ?0.05. The magnitude of parameter expression is color-coded with red for a relative increase in expression and blue for a relative decrease in expression. CM CD4+T cell, central memory CD4+T cell; EM CD4+T cell, effector memory CD4+T cell; CM CD8+T cell, central memory CD8+T cell; EM CD8+T cell, effector memory CD8+T cell; Th cell, helper T cell; Tfh cell, follicular helper T cell; Tc cell, cytotoxic T lymphocyte; Treg cell, regulatory T cell; Breg cell, regulatory B cell T lymphocyte The percentage of CD4+ T cells at different stages of differentiation were calculated, and significant differences between the AS patients and HCs are shown in Fig.?2. CCR7+ CD4+T cells including na?ve CD4+T cells (CD3+CD4+CD45RA+CCR7+, Fig. ?Fig.2a)2a) and central memory CD4+T cells (CD3+CD4+CD45RA?CCR7+, Sorbic acid Fig.?2c) were significantly increased in the AS group, but CCR7? CD4+T cells including terminally differentiated CD4+T cells (CD3+CD4+CD45RA+CCR7?, Fig.?2b), and effector memory CD4+T cells (CD3+CD4+CD45RA?CCR7?, Fig.?2d) were significantly decreased. Open in a separate window Fig. 2 Differences in CD4+ T cells and CD8+ T cells in the AS and HC groups at different stages of differentiation. value summary: *value summary: *value summary: *value summary: * em P /em ? ?0.05, ** em P /em ? ?0.01, *** em P /em ? ?0.001, **** em P /em ? ?0.0001. Treg cell, regulatory T cell; Tc cell, cytotoxic T lymphocyte; Tfh cell, follicular helper T cell; B10 cell, IL-10 producing regulatory B cell The number of regulatory lymphocytes detected in the blood of the AS patients changed significantly after Anbainuo treatment, with the percentage of Treg FLJ20315 cells (CD3+CD4+CD25+CD127?, Fig.?5b) and B10 cells (CD3?CD19+CD24+CD27+ CD38?IgD+IgM+, Fig.?5c) increasing significantly but immature Bregs (CD3?CD19+CD24+ CD27CD38+IgD+IgM+, Fig.?5g) decreasing significantly. Simultaneously, we measured the number of Th cells (Th1 cells, Th2 cells, Th17 cells), Tc cells (Tc1 cells, Tc2 cells, and Tc17 cells), and Tfh cells (Tfh1 cells, Tfh2 cells, and Tfh17 cells) before and after Anbainuo therapy. As shown in Fig.?5, the proportion of Tc1 cells (CD3+CD8+CXCR3+CCR4?CXCR5?, Fig.?5e) decreased, and the proportion of Tfh17 cells (CD3+CD4+CXCR3?CCR4?CXCR5+ CCR6+, Fig.?5f) increased after treatment. However, apart from immature Bregs and B10 cells, the proportion of various B cell subtypes did not change significantly after treatment with Anbainuo. Correlation between immune cells and disease activity In order to understand whether disease activity of AS patients is related to immune cell imbalance, we analyzed the correlation between disease activity indicators (CRP and ASDAS) and frequency of immune cells. But only the frequency of Tc1 cells (CD3+CD8+CXCR3+CCR4?CXCR5?) was found to be negatively correlated with CRP level ( em r /em ?=???0.182, em P /em ?=?0.041). To understand the correlation between changes in disease status (including CRP, BASDAI, and ASDAS) and changes in lymphocyte frequency after Anbainuo therapy, Spearmans rank correlation analyses showed that the decrease in CRP was positively correlated with the increase in the frequency of Tregs (CD3+CD4+CD25+CD127?) following Anbainuo therapy Sorbic acid for 12?weeks ( em r /em ?=?0.489, Sorbic acid em P /em ?=?0.018). Discussion As we know, the onset of AS suffers from the relationship between the host genetics, the intestinal microbiome, and the immune response . AS has long been associated with inheritance of the HLA allele B27 , and the pathogenic role of HLAB27 remains unclear despite intensive research. The arthritogenic peptide theory proposes that HLAB27 plays a central pathogenic role in the presentation of joint-specific peptides to CD8+ cytotoxic T cells..