ABSTRACT
Objective To identify interleukin 17 (IL-17) and B cell activating factor (BAFF) that could influence B cell biology by detecting the expression of BAFF in the serum and labial salivary glands from primary Sj(o)gren's syndrome (pSS) patients and to test the apoptosis rates of B cells cultured with Th17 cells which were transfected with IL-17-siRNA,BAFF-siRNA.Methods A total of 40 patients with pSS who were referred to the Department of Rheumatology and Immunology at Anhui Provincial Hospital from June 2011 to June 2012 were enrolled into this study.The expression of BAFF on salivary gland and serum from pSS patients and healthy controls were detected by ELISA and immunohistochemical examination (22 patients with pSS).Flow cytometry was used to detect B cell's apoptosis,BAFF and IL-17 interfered with amplified Th17 cells,and co-cultured with B cells.Immunoblot was used to detect supernatant antibody in 5 patients with pSS.Independent samples t test was used for statistical analysis.Results In all pSS specimens,infiltrating inflammatory cells expressed BAFF,so did some ductual cells,but acinar cells did not express these markers.There was no expression of BAFF in the controls.BAFF-positive cell numbers in the labial salivary glands of pSS patients with focal infiltrating lymphocytes were more than that with non-focal infiltrating lymphocytes (888±372 vs 164±161,t=5.94,P＜0.05),and the percentage of BAFF-positive lymphocytes over the total infiltrating lymphocytes in the salivary glands of pSS patients with focal infiltrating lymphocytes [(0.18 ±0.08) ％] was higher than those with non-focal infiltrating lymphocytes [(0.09 ±0.07) ％] (t =3.03,P＜0.05).The level of soluble BAFF in patients with pSS [(6.0±2.8) ng/ml] was significantly higher than the controls [(3.8±1.7) ng/ml,t=3.26,P＜0.05].BAFF or IL-17 transfected group,B cell apoptosis rate [(24± 5)％,(23±5)％] were significantly higher than the non-transfected group [(7±4)％],t=4.6,4.4; P＜0.05].And there was no significant difference when compared with cultured B cells (P＞0.05).Compared with the controls,no antibody could be detected in the supernatants.Conclusion BAFF may be involved in the process of local inflammatory damage of the pSS,it may have a synergistic effect with IL-17 on abnormal B cell function.
ABSTRACT
Objective To investigate the frequencies of CD4+CXCR5+T cells in the CD4+T cells of peripheral blood of patients with systemic lupus erythematosus (SLE) and the effect of glucocorticoid on it.Methods Frequencies of CD4+CXCR5+T cell were analyzed by flow cytometry in 45 active,20 inactive SLE patients and 20 healthy controls.Differences between groups and the effect of glucocorticoid were analyzed.Meanwhile, the expression of CXCR5 on CDI9+B cells was analyzed. Independent sample t test was used for statistical analysis between twogroups, ANOVA was applied for data analysis between 3 groups,,nonparameterical Spearman's analysis was used for correlation analysis and repeated measurement ANOVA were used to compare the parameters before and after treatment. Results The percentage of CD4+CXCR5+ in CD4+T cells was increased in patients with SLE compared with healthy controls[(16±7)% vs (12±3)%, P＜0.01].It was increased in patients with active SLE [(18±7)%] compared with healthy controls (P＜0.05) but there was no significant difference between inactive SLE[(11±4)%] and healthy controls(P＞0.05). The percentage in patients with LN was higher than that in patients without LN, but without significant difference[(18±7)%vs (14±7)%, P=0.05 ]. The percentage of CD4+CXCR5+T cells was positively correlated with SLEDAI,the titer of ANA and level of ESR but negatively correlated with the level of C3 (P<0.05 for each).No correlation was found between duration and the levels of CRP and immunoglobulin.. The percentage in patients with high anti-dsDNA group was also higher than that of the low group, but no differences were found between anti-Sm antibody positive and negative groups neither between anti-SSA/SSB antibody positive and negative groups(P＞0.05 for each).The expression level of CXCR5 on CD19+B cells in active SLE patients was lower than that of healthy controls[(85±11)% vs (94±3)%, P＜0.05 ]. The percentages of CD4+CXCR5+T cells in 10 untreated active SLE patients were decreased at day 1,day 3 and day 7 after being treated with dexamethasone (20mg/d) when compared with those before the treatment (P＜0.05 for each), but the percentages of CD19+CXCR5+B cells had no significant change (P＞0.05 for each).Conclusion These results demonstrate that the abnormality of CD4+CXCR5+T cells may play an important role in the pathogenesis of SLE.