摘要
目的 研究HLA DRβ1特异性非T细胞结合肽对胶原性关节炎的抑制作用 ,探索类风湿关节炎新的免疫治疗方法。方法 以丙氨酸及甘氨酸替换Ⅱ型胶原 (CⅡ ) 2 6 3 2 72多肽中与T细胞受体结合的氨基酸 ,利用HLA DR1转基因抗原呈递细胞及CⅡ特异性T细胞组成的激活体系 ,从这些CⅡ 2 6 3 2 72修饰肽中筛选非T细胞结合肽。牛CⅡ +完全福氏佐剂诱导大鼠胶原性关节炎 (CIA)动物模型 ,应用不同剂量 (10、5 0、10 0 μg)的非T细胞结合肽 2 6 8A皮下注射进行为期 16d的治疗 ,观察大鼠关节肿胀的变化情况。酶联免疫吸附法 (ELISA)检测外周血CⅡ抗体和肿瘤坏死因了 (TNF) α浓度 ,HE染色检测CIA病理变化。结果 7条CⅡ 2 6 3 2 72修饰肽中以丙氨酸替换第 2 6 7、2 6 8、2 6 9、2 70位残基的 4条肽 (2 6 7A、2 6 8A、2 6 9A及 2 70A)和用丙氨酸和甘氨酸连续替换 2 6 8、2 6 9、2 70位残基的 1条肽 (Sub2 6 8 2 70 )对T细胞激活能力显著低于原型肽 (P <0 0 1)。利用 2 6 8A多肽对CIA进行治疗的结果发现 ,3个治疗组CIA缓解率显著高于对照组 (P <0 0 5或P <0 0 1) ;3个治疗组的关节炎评分显著低于对照组 (P <0 0 5或P <0 0 1)。 3个治疗组TNF α浓度及CⅡ抗体浓度均显著低于对照组 (P <0 0 5或P <0 0 1)。
Objective To evaluate the inhibitory effect of modified CⅡ 263 272 peptide with substitutions of TCR binding residues on collagen induced arthritis (CIA),and explore therapeutic strategy of rheumatoid arthritis.Methods A panel of modified peptides was synthesised with substitutions of TCR binding residues of CⅡ 263 272 with glycine and alanine.Hyporesponsive peptides(non T cell binding peptides) were selected via T cell proliferation assay.CIA model was induced by intradermal injection of bovine CⅡ 400 μg+complete Freund′s adjuvant.Modified CⅡ 263 272 peptides 268A was injected subcutaneously in different doses (10,50,100 μg,respectively) after onset of CIA.Subsequently the swollen joint score was recorded every day to evaluate the severity change of arthritis.TNF α and CⅡ antibodies in serum were examined by ELISA.Joint synovial HE staining was preformed after mouse sacrifice.Results Modified CⅡ 263 272 peptides with individual substitution of residues 267(Q),268(G),269(P),270(K) or consecutive substitution of 269,269,270 did not or barely stimulate T cell proliferation,compared to wild type CⅡ ( P <0 01).A significant higher response rate was found in the peptide receiving group compared with control after treatment ( P <0 05 or P <0 01,respectively).Compared with control group,the swollen joints scores were much lower in the treatment group on 10 days after modified CⅡ peptied administration ( P <0 05 or P <0 01,respectively).Concentration of TNF α and CⅡ antibody in 3 treatment groups are significantly lower than in control ( P <0 05 or P <0 01 respectively).Conclusion The non T cell binding peptides are hyporesponsive in T cell activation and show inhibitory effect on CIA.It may be a possible strategy in treatment of RA.
出处
《中华风湿病学杂志》
CAS
CSCD
2002年第6期397-401,共5页
Chinese Journal of Rheumatology
基金
国家杰出青年基金资助项目 (3 0 0 2 5 0 40 )
北京大学 985资助项目
