鼠源T细胞受体单克隆抗体对Ⅱ型胶原诱导DBA/1小鼠关节炎的治疗作用

Effect of mouse monoclonal antibody against T cell receptor oncollagen type Ⅱ-induced arthritis in DBA/1 mice

目的观察鼠源T细胞受体(mTCR)单抗对类风湿性关节炎的治疗作用。方法将DBA/1小鼠随机分为正常对照、模型对照、地塞米松0.5 mg·kg-1和mTCR单抗25 mg·kg-1治疗组,每组6只。除正常对照组外,其余各组小鼠分别在第1天和第22天皮内注射牛Ⅱ型胶原与完全弗氏佐剂的乳化剂,制备Ⅱ型胶原诱导的关节炎(CIA)小鼠模型。地塞米松治疗组于第29～41天皮下注射地塞米松磷酸钠注射液,隔日一次;mT-CR单抗组分别在第0天和第21天皮下注射mTCR单抗各1次;正常和模型对照组皮下注射等体积生理盐水。动态观察小鼠体质量、足掌厚度和关节炎指数的变化,并计算CIA的发生率。给药后第60天实验结束,处死小鼠,HE染色观察踝关节组织病理变化,多功能流式液相芯片分析仪测定小鼠血清γ干扰素(IFN-γ),肿瘤坏死因子α(TNF-α)和白细胞介素4(IL-4)的含量。结果在第42天,模型组小鼠CIA发生率为6/6,地塞米松和mTCR单抗治疗组分别为2/6和1/6;第49天,模型组和地塞米松治疗组CIA的发生率仍分别为6/6和2/6,mTCR单抗治疗组升高为5/6。在整个发病过程中,地塞米松和mTCR单抗治疗组小鼠的足掌厚度和关节炎指数较模型组降低(P<0.05,P<0.01),其中第38天时,正常对照组、模型组、地塞米松和mTCR单抗治疗组小鼠足掌厚度分别为2.34±0.06,2.96±0.49,2.61±0.44和(2.29±0.13)mm;关节炎指数分别为0.0±0.0,4.0±2.6,0.7±1.0和0.3±0.8。实验结束时,地塞米松和mTCR单抗治疗组血清IL-4含量分别为94±12和(105±38)ng·L-1,与模型组(67±8)ng·L-1比较明显升高(P<0.05);TNF-α含量分别为4.8±0.7和(3.8±0.8)ng·L-1,与模型组(6.7±1.5)ng·L-1比较明显降低(P<0.05);各组IFN-γ含量无明显变化。组织病理观察结果显示,与模型组相比,地塞米松和mTCR单抗治疗组小鼠踝关节炎症细胞浸润明显减轻,骨破坏减弱。结论 mTCR单抗对类风湿性关节炎可能具有治疗作用。

OBJECTIVE To investigate the effect of mouse monoclonal antibody against T cell re- ceptor （mTCR mAb） on rheumatoid arthritis. METHODS Male DBA/1 mice were randomly divided into normal control, type Ⅱ collagen-induced arthritis （CIA） model, dexamethasone and mTCR mAb treated groups. To induce the mouse CIA model, 100 pl of bovine type Ⅱ collagen and complete Freund adju- vant mixture emulsion was intracutaneously injected into each mouse on the 1st and 22nd day. Dexam- ethamoson 0.5 mg· kg -1 was sc given to CIA mice on the 29th to 41 st day, once every two days. mTCR mAb 25 mg· kg-1 was sc given on the 0 （before model preparation） and 21st day. The body mass, foot palm thickness, joint inflammatory index and pathological changes of each mouse were abserved. On the 60th day the experiment was finished, and the serum level of interferon-γ（ IFN-γ） , tumor necrosis factor-α（TNF-α） and interleukin-4（IL-4） was measured using Luminex200 LCD chip analysis platform. RESULTS On the 42nd day, the incidence of CIA in the model, dexamethasone and mTCR mAb groups was 6/6, 2/6 and 1/6, respectively. On the 49th day, the incidence in mTCR mAb group reached 5/6 but remained unchanged in the other groups. Compared with model group, the mice of dex- amethasone and mTCR mAb groups showed improvement of foot palm thickness, joint inflammatory index and pathological changes. On the 38th day, the thickness of paws in normal control, model, dexa- methasone and mTCR mAb groups was 2.34 ±0.06, 2.96±0.49, 2.61 ±0.44 and （2.29±0.13）mm, respectively, and the joint inflammatory index was 0.0 ±0.0, 4.0 ±2.6, 0.7 ±1.0 and 0.3 ±0.8. At the end of the experiment, the serum level of IL-4 in normal control, model, dexamethasone and mTCR mAb groups was 106 ±7, 67 ±8, 94 ±12 and （105 ±38） ng·L-1 , TNF-α was 0.8 ±0.3, 6.7 ±1.5, 4.8 ±0.8 and （3.8±0.8）ng·L-1 and IFN-7. was 2.0 ±0.0, 2.3 ±0.6, 2.3 ±0.6 and （2.8 ±1.9）ng·L-1. CONCLUSION mTCR mAb exerts therapeutic effect on rheumatoid arthritis.