重组人Ⅱ型胶原250-270多肽对类风湿关节炎患者外周血T细胞的活化作用  

Enhanced T cell proliferative response to recombinant human type Ⅱ collagen peptide 250-270 in patients with rheumatoid arthritis

作　　者：李学义[1] 朱平[1] 樊春梅[1] 王彦宏[1] 刘玲[1] 史战国[1] 

机构地区：[1]第四军医大学西京医院临床免疫科,西安710032

出　　处：《中华风湿病学杂志》2003年第2期75-78,共4页Chinese Journal of Rheumatology

基　　金：国家重点科技攻关资助项目 (96 90 1 0 5 2 62 )

摘　　要：目的　研究重组人Ⅱ型胶原 2 5 0 2 70多肽 (recombinanthumancollagentypeⅡ pep tide 2 5 0 2 70 ,rhCⅡ 2 5 0 2 70 )对类风湿关节炎 (RA)患者外周血特异性T细胞的活化作用 ,为口服耐受治疗RA提供研究材料和实验依据。方法　不同浓度 (0 1、1 0、10、10 0、10 0 0 μg ml)的 6聚rhCⅡ 2 5 0 2 70体外刺激RA患者外周血单个核细胞 (PBMC) ;等浓度的融合表达的基础蛋白 (GST)和磷酸盐缓冲盐水 (PBS)作为阴性对照 ;化学合成CⅡ 2 5 0 2 70和十四烷酰佛波醇乙酯 (PMA)刺激组为阳性对照 ,体外培养 1～ 6d。流式细胞仪 (FCM)测定刺激前后各组培养细胞中T(CD3+ )淋巴细胞表面活化标志CD2 5的表达、5 溴脱氧尿嘧啶 (BrdU)在DNA中的掺入及细胞因子干扰素 (IFN) γ和白细胞介素 (IL) 4的分泌 ,以验证其对RA患者外周血特异性T细胞的活化作用。结果　rhCⅡ2 5 0 2 70合适的刺激浓度为 10 0 μg ml,在此浓度下刺激 4d ,RA患者外周血T细胞中CD2 5 + 、Br dU+ 细胞率分别为 (17± 4 ) %、(35± 5 ) % ,均显著高于同行对照GST组 (P <0 0 1) ,而和化学合成CⅡ 2 5 0 2 70刺激组差异无显著性 (P >0 0 5 )。同等条件下rhCⅡ 2 5 0 2 70刺激组T细胞中IL 4、IFN γ阳性细胞率分别为 (13 3± 4 0 ) %、(6 0± 2 1)Objective To investigate the activation of T cells from peripheral blood mononuclear cells (PBMC) in patients with rheumatoid arthritis (RA) induced by recombinant human type Ⅱ collagen peptide 250270 (rhCⅡ 250270).Methods The proliferation of T cells from RA patients,including CD25 expressed,DNA synthesized and IL 4 or IFN γ produced in responses to rhCⅡ (0 1,1 0,10,100 and 1 000 μg/ml),synthetic CⅡ 250270 (10 μg/ml) and GST protein were evaluated by flow cytometry system (FCM) on different time points (1～6 d).Results The results of flow cytometry indicated that 100 μg/ml of rhCⅡ 250270,cultured for 4 d was the optimal condition to activate T cells in vitro .The frequency of CD25 + and BrdU + T cells in PBMC from RA patients stimulated by rhCⅡ 250 270 were (17±4)% and (35±5)% respectively,which were significantly higher than those of GST′s groups,however,no significant difference was detected between rhCⅡ 250270 and synthesized CⅡ 250270 groups.The percentage of IL 4 and IFN γ produced cells in PBMC induced by rhCⅡ 250270 was much higher [(13 3±4 0)% and (6 0±2 1)%,respectively] compared with GST groups ( P <0 01).Conclusion The results demonstrate that the rhCⅡ 250270,as well as synthesized CⅡ 250270 could promote T cell proliferation from RA patients in vitro .It may provide a material for further researching the pathogenesis and therapy of rheumatoid arthritis.

关 键 词：类风湿关节炎 外周血T细胞 活化 多肽 重组人Ⅱ型胶原 

分 类 号：R593.22[医药卫生—内科学]