强直性脊柱炎患者外周血中调节性B细胞亚型的表达及其与疾病活动度的关系  

Frequencies of human regulatory B cells in PBMC in ankylosing spondylitis patients and its clinical significance

作　　者：薛国辉[1] 华琳[1] 刘晓峰[1] 陈雪礼[1] 董亮[2] 潘京[1] 周南进[3,4] 

机构地区：[1]九江市第一人民医院/南昌大学附属九江医院检验科,江西九江332000 [2]九江市第一人民医院/南昌大学附属九江医院风湿免疫科,江西九江332000 [3]江西省医学科学研究院,南昌330006 [4]南昌大学免疫与生物治疗研究所,南昌330006

出　　处：《临床检验杂志》2015年第9期662-667,共6页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金(81160056)

摘　　要：目的探讨强直性脊柱炎(ankylosing spondylitis,AS)中调节性B细胞(regulatory B cells,Breg)的水平及与疾病活动度间的关系。方法选取确诊AS患者38例,按BASDAI评分分为活动期21例和非活动期17例,同时选取30例健康人对照者,AS患者与对照组间一般资料差异无统计学意义。ASDAS-CRP和BASDAI评分作为疾病活动度评价指标。用流式细胞术检测AS患者外周血CD19+CD24hiCD38hiBreg、CD19+CD24hiCD27+Breg、CD4+CD25highTreg及CD3+IL-17+CD8-TH17比例,荧光定量PCR检测RORγt和Foxp3 mRNA表达水平。结果 AS患者CD19+CD24hiCD38hiBreg比例为(2.10±0.16)%,显著低于对照组(3.44±0.80)%,其中活动期患者[(1.88±0.66)%]低于非活动期患者[(2.36±0.57)%],均低于对照组,差异均有统计学意义。AS患者CD19+CD24hiCD38hiBreg与Foxp3 mRNA成显著正相关性(r=0.369,P<0.05),与TH17、RORγt mRNA、BASDAI评分及ASDAS-CRP负相关(r分别为-0.338、-0.661、-0.411、-0.714,P均<0.05),而与Treg无相关性(r=-0.006,P>0.05)。而AS患者中CD19+CD24hiCD27+Breg亚群比例与对照组间差异则无统计学意义,但活动期患者比例有所降低,与非活动期患者及对照组比较,差异有统计学意义。AS患者中CD19+CD24hiCD27+Breg亚群比例与Foxp3 mRNA正相关(r=0.344,P<0.05),与ASDAS-CRP负相关(r=-0.383,P<0.05),但与TH17、Treg、RORγt mRNA及BASDAI评分均无显著相关性。结论 AS患者中存在异常的Breg表型主要为CD19+CD24hiCD38hiBreg,其在AS患者中显著降低,并与疾病活动度密切相关,且与RORγt和Foxp3 mRNA水平及TH17数量存在一定的关联。Objective To determine the frequencies of regulatory B cells（ Breg） in peripheral blood mononuclear cell（ PBMC） in ankylosing spondylitis（ AS） patients and its clinical association with AS. Methods Thirty-eight consecutive patients with AS were included in this study. Based on BASDAI scores,AS patients were divided into active group（ n = 21） and inactive group（ n = 17）. Thirty age- and sex-matched healthy individuals served as controls. ASDAS-CRP and BASDAI were used as indicators of AS disease activity.Flow cytometry was used to determine the frequencies of CD19＋CD24hiCD38hiBreg,CD19＋CD24hiCD27＋Breg,CD4＋CD25highTreg and CD3＋IL-17＋CD8-TH17 cells in the AS patients,and mRNA levels of RORγt and Fox P3 were examined by quantitative real-time PCR.The patients＇ perceived disease activity was recorded by BASDAI and ASDAS using serum CRP levels（ ASDAS-CRP）. Results The percentage of CD19＋CD24hiCD38hiBreg in AS patients [（ 2. 10 ± 0. 16%） ] was lower than that in healthy control [（ 2. 10 ± 0. 16） % vs（ 3. 44 ±0. 80） %]. Among AS patients,higher proportion of CD19＋CD24hiCD38hiBreg was observed in active patients compared with inactive patients [（ 1. 88 ± 0. 66） % vs（ 2. 36 ± 0. 57） %]. CD19＋CD24hiCD38hiBreg ratio was significantly and positively correlated with the levels of Foxp3 mRNA（ rFoxp3 mRNA= 0. 369,P〈0. 05）,and negatively related to TH17,RORγt mRNA,BASDAI and ASDASCRP socre（ rTH17=- 0. 338,P〈0. 05; rRORγt mRNA=- 0. 661,P〈0. 01; rBASDAI=- 0. 411,P〈0. 05; rASDAS-CRP=- 0. 714,P〈0. 01）,while they were slightly correlated with Treg（ rTreg=- 0. 006,P〈0. 05）. However,the frequency of CD19＋CD24hiCD27＋Breg,another type of defined Breg,did not show significantly different between AS patients and control. but the propotion of CD19＋CD24hiCD27＋Breg in active AS patients was lower than that in control or inactive cases,and the differences were statistically significant. In AS group,the frequency of CD19＋CD24hiCD27＋Breg wa

关 键 词：调节性B细胞 强直性脊柱炎 调节性T细胞 T辅助细胞17 

分 类 号：R446.6[医药卫生—诊断学]