合成环瓜氨酸化蛋白短肽的免疫原性和致关节炎性研究  

Study on immunogenicity and arthritogenicity of a synthetic cyclic citrullinated peptide

作　　者：陈恩生[1,2] 崔明珠[1] 赵晓峰[3] 余焙佳[1] 肖长虹[1] 顾为望[2] CHEN En-Sheng CUI Ming-Zhu ZHAO Xiao-Feng YU Bei-Jia XIAO Chang-Hong GU Wei-Wang.(TCM-Integrated Hospital of Southern Medical University, Guangzhou 510515, China)

机构地区：[1]南方医科大学中医药学院,广州510515 [2]南方医科大学实验动物中心(比较医学研究所),广州510515 [3]南方医科大学中西医结合医院,广州510515

出　　处：《中国免疫学杂志》2017年第1期25-30,共6页Chinese Journal of Immunology

基　　金：广东省科技计划项目(No.2014A030304025)

摘　　要：目的:构建环瓜氨酸化蛋白短肽诱导性小鼠关节炎模型,探讨该短肽的免疫原性、致关节炎性。方法:36只DBA/1小鼠随机分为3组,于第0天、第21天分别以Ⅱ型胶原(CⅡ,CIA)、环瓜氨酸化波形蛋白短肽(CCit-Vim,CCV-IA)和偶联血蓝蛋白(KLH)的环瓜氨酸化波形蛋白短肽(CCit-Vim+KLH,CCV+K-IA)皮下注射。ELISA检测血清抗CⅡ抗体、抗CCit-Vim抗体、抗CCP抗体及TNF-α,间接免疫荧光法(IIF)检测抗大鼠食管角蛋白抗体(AKA),对关节炎指数(AI)、足容积、踝关节病理学进行评价。结果:CCV+K-IA小鼠关节炎发病率为25%(3/12),但关节炎出现时间晚,持续时间短,发病率及关节炎程度均低于CIA;CCV-IA无关节炎发生。CCV+K-IA产生抗CCit-Vim抗体高于CIA(P=0.031),产生抗CCP抗体反而低于CIA(P=0.007)。CCV+K-IA、CCV-IA产生的抗CⅡ抗体水平均低于CIA(P<0.05)。CCV+K-IA与CIA的TNF-α高于CCV-IA(P<0.05)。CCV+K-IA的AKA阳性率高于其余两组(50%vs CCV-IA 25%、CIA 16.67%)。CCV-IA和CCV+K-IA踝关节病理显示轻度滑膜增生,无滑膜血管翳形成及炎性细胞浸润。结论:偶联KLH的CCit-Vim短肽不仅具有强的免疫原性,而且具有致关节炎性;与CⅡ比较,CCit-Vim+KLH能诱导出更高的AKA阳性率。Objective： To establish a synthetic cyclic citrullinated peptide induced arthritis model in mice,explore immunogenicity and arthritogenicity of this peptide. Methods： 36 DBA /1 mice were randomly divided into three groups,which were injected the type Ⅱ collagen（ CⅡ,CIA） emulsion,cyclic citrullinated vimentin peptide（ CCit-Vim,CCV-IA） emulsion,cyclic citrullinated vimentin peptide conjugated KLH（ CCit-Vim ＋ KLH,CCV ＋ K-IA） emulsion on day 0 and 21,respectively. Using arthritis index（ AI）,paw swelling to evaluate the incidence of arthritis; ELISA tested serum anti-CCit-Vim antibody,anti-CⅡantibody,anti-CCP antibody and TNF-α,IIF detected AKA; Histopathology of the ankle joint was obsearved. Results： There were three mice appeared arthritis in CCV ＋ K-IA,the incidence rate of 25%,but arthritis occurs later time,short duration,and the incidence and extent of arthritis were lower than the CIA. CCV-IA no arthritis performance. CCV ＋ K-IA produce anti-CCit-Vim antibody were significantly higher than those in CIA（ 1. 32± 0. 59 vs 0. 78 ± 0. 27,P = 0. 031）. While Anti-CCP antibody of CCV ＋ K-IA were significantly lower than CIA（ 54. 73 ± 7. 33 vs64. 37 ± 9. 91,P = 0. 007）. The anti-CⅡ antibody in CCV ＋ K-IA and CCV-IA were lower than the CIA（ 15. 73 ± 2. 10,16. 71 ± 3. 03 vs 19. 50 ± 2. 36,P〈0. 05）. The TNF-α produced by CCV ＋ K-IA and CIA were both significantly higher than the CCV-IA（ 645. 61 ±35. 26,618. 98 ± 53. 32 vs 533. 63 ± 79. 49,P〈0. 05）. The AKA positive rate of CCV ＋ K-IA is 50%（ 6 /12）,significantly higher than CCV-IA 25%（ 3 /12） and CIA 16. 67%（ 2 /12）. Histopathology of the ankle showed that the CCV-IA and CCV ＋ K-IA have a mild synovial hyperplasia,no obvious synovial pannus formation and inflammatory cell infiltration. Conclusion： The cyclic citrullinated peptide conjugated KLH not only has stronger immunogenicity but also has arthritogenicity. It induced a higher positive rate of AKA than C Ⅱ.

关 键 词：类风湿关节炎 瓜氨酸化短肽 胶原诱导性关节炎 动物模型 

分 类 号：R392.9[医药卫生—免疫学]