[改变的CII263-272肽的黏膜给药抑制胶原诱导的关节炎]

[Mucosal administration of altered CII263-272 peptide inhibits collagen-induced arthritis].

作者信息

Zhao Jin Xia, Li Ru, He Jing, Shi Jin Xia, Li Zhan Guo

机构信息

Department of Rheumatology & Immunology, Peking University People's Hospital, Beijing, China.

出版信息

Beijing Da Xue Xue Bao Yi Xue Ban. 2008 Dec 18;40(6):562-6.

PMID:19088823

Abstract

OBJECTIVE

To evaluate the effect of mucosal administration of altered collagen II(CII)263-272 peptide (267Q-->A, 270K-->A and 271G-->A) on collagen induced arthritis (CIA), and to explore the mechanism of the inhibitory effect of the altered CII263-272 peptide on CIA.

METHODS

CIA was induced in Lewis rats by immunization with bovine CII. Altered CII263-272 peptide was given intranasally beginning from the onset of arthritis (100 microg/dose, daily for 5 doses and continuing every other day for other 7 doses). Wild CII263-272 peptide (100 microg/dose) or PBS was administered as controls with the same procedure. Therapeutic effects were evaluated by arthritis scores, body weight change, and joint pathologic scores. The anti-CII antibody and its subtypes were measured with ELISA. The cytokines of IFN-gamma and IL-10 were measured with ELISA. The induction of regulatory T cells was assessed by FACS analysis of percentage of peripheral CD4(+)CD25(+) T cells, and by real-time PCR analysis of the expression of Foxp3 and TGF-beta mRNA.

RESULTS

(1) Following treatment with the altered CII263-272 peptide, arthritis scores were reduced and body weight was increased. The mean arthritis scores of rats treated with altered peptide, wild peptide and PBS were 2.50 +/- 2.43, 4.50 +/- 2.23 and 6.33 +/- 2.73, respectively. The altered peptide could retard the histologic lesion of the joints. (2) The titers of anti-CII antibodies IgG and IgG1 in the three groups were similar, but the IgG2a in altered peptide-treated rats decreased markedly as compared with PBS-treated rats (0.56 +/- 0.19 vs 0.95 +/- 0.29, P<0.05). The serum IFN-gamma in rats treated with altered peptide, wild peptide and PBS were (185.33 +/- 29.77), (231.62 +/- 41.82) and (220.64 +/- 83.61) ng/L, respectively (P<0.05). No difference was found in the levels of serum IL-10 among the three groups. (3) There was no significant difference in the percentage of peripheral CD4(+)CD25(+) T cells and the expression level of Foxp3 and TGF-beta mRNA.

CONCLUSION

Mucosal administration of altered CII263-272 peptide could effectively inhibit the progression of CIA. It can decrease the IgG2a subtype of anti-CII antibodies and IFN-gamma, and inhibit Th1 response in vivo. Altered C II263-272 peptide may be therapeutic for RA.

摘要

目的

评估经黏膜给予改变的Ⅱ型胶原蛋白（CII）263 - 272肽（267Q→A、270K→A和271G→A）对胶原诱导性关节炎（CIA）的影响，并探讨改变的CII263 - 272肽对CIA抑制作用的机制。

方法

通过用牛CII免疫Lewis大鼠诱导CIA。从关节炎发作开始经鼻给予改变的CII263 - 272肽（100μg/剂量，每日1次，共5次剂量，之后隔日1次，共7次剂量）。以相同程序给予野生型CII263 - 272肽（100μg/剂量）或PBS作为对照。通过关节炎评分、体重变化和关节病理评分评估治疗效果。用ELISA法检测抗CII抗体及其亚型。用ELISA法检测IFN - γ和IL - 10细胞因子。通过流式细胞术分析外周血CD4(+)CD25(+) T细胞百分比以及通过实时PCR分析Foxp3和TGF - β mRNA表达来评估调节性T细胞的诱导情况。

结果

（1）用改变的CII263 - 272肽治疗后，关节炎评分降低，体重增加。用改变的肽、野生肽和PBS治疗的大鼠平均关节炎评分分别为2.50±2.43、4.50±2.23和6.33±2.73。改变的肽可延缓关节的组织学损伤。（2）三组中抗CII抗体IgG和IgG1的滴度相似，但与PBS治疗的大鼠相比，改变的肽治疗的大鼠中IgG2a明显降低（0.56±0.19对0.95±0.29，P<0.05）。用改变的肽、野生肽和PBS治疗的大鼠血清IFN - γ分别为（185.33±29.77）、（231.62±41.82）和（220.64±83.61）ng/L（P<0.05）。三组血清IL - 10水平无差异。（3）外周血CD4(+)CD25(+) T细胞百分比以及Foxp3和TGF - β mRNA表达水平无显著差异。