Zhu Ping, Li Xiao-Yan, Wang Hong-Kun, Jia Jun-Feng, Zheng Zhao-Hui, Ding Jin, Fan Chun-Mei
Department of Clinical Immunology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, Shaanxi Province, PR China.
Clin Immunol. 2007 Jan;122(1):75-84. doi: 10.1016/j.clim.2006.08.004. Epub 2006 Oct 11.
Oral antigen is an attractive approach for the treatment of autoimmune and inflammatory diseases. Establishment of immune markers and methods in evaluating the effects of antigen-specific cellular and humoral immune responses will help the application of oral tolerance in the treatment of human diseases. The present article observed the effects of chicken collagen II (CII), the recombinant polymerized human collagen II 250-270 (rhCII 250-270) peptide and synthesized human CII 250-270 (syCII 250-270) peptide on the induction of antigen-specific autoimmune response in rheumatoid arthritis (RA) peripheral blood mononuclear cells (PBMC) and on the specific cellular and humoral immune response in collagen-induced arthritis (CIA) and mice fed with CII (250-270) prior to immunization with CII. In the study, proliferation, activation and intracellular cytokine production of antigen-specific T lymphocytes were simultaneously analyzed by bromodeoxyuridine (BrdU) incorporation and flow cytometry at the single-cell level. The antigen-specific antibody and antibody-forming cells were detected by ELISA and ELISPOT, respectively. CII (250-270) was found to have stimulated the response of specific lymphocytes in PBMC from RA patients, including the increase expression of surface activation antigen marker CD69 and CD25, and DNA synthesis. Mice, fed with CII (250-270) before CII immunization, had significantly lower arthritic scores than the mice immunized with CII alone, and the body weight of the former increased during the study period. Furthermore, the specific T cell activity, proliferation and secretion of interferon (IFN)-gamma in spleen cells were actively suppressed in CII (250-270)-fed mice, and the serum anti-CII, anti-CII (250-270) antibody activities and the frequency of specific antibody-forming spleen cells were significantly lower in CII (250-270)-fed mice than in mice immunized with CII alone. These observations suggest that oral administration of CII (250-270) can suppress the cellular and humoral immune response in collagen-induced arthritis, and the simultaneous analysis of antigen-specific cellular and humoral immune responses at single-cell level will help the understanding of the oral tolerance mechanisms in CIA and the development of innovative therapeutic intervention for RA.
口服抗原是治疗自身免疫性疾病和炎症性疾病的一种有吸引力的方法。建立免疫标志物以及评估抗原特异性细胞免疫和体液免疫反应效果的方法，将有助于口服耐受在人类疾病治疗中的应用。本文观察了鸡胶原蛋白II（CII）、重组聚合人胶原蛋白II 250 - 270（rhCII 250 - 270）肽和合成人CII 250 - 270（syCII 250 - 270）肽对类风湿关节炎（RA）外周血单个核细胞（PBMC）中抗原特异性自身免疫反应的诱导作用，以及对胶原诱导性关节炎（CIA）和在以CII免疫前喂食CII（250 - 270）的小鼠中特异性细胞免疫和体液免疫反应的影响。在该研究中，通过溴脱氧尿苷（BrdU）掺入和流式细胞术在单细胞水平同时分析抗原特异性T淋巴细胞的增殖、活化和细胞内细胞因子产生情况。分别通过ELISA和ELISPOT检测抗原特异性抗体和抗体形成细胞。发现CII（250 - 270）可刺激RA患者PBMC中特异性淋巴细胞的反应，包括表面活化抗原标志物CD69和CD25的表达增加以及DNA合成。在以CII免疫前喂食CII（250 - 270）的小鼠，其关节炎评分显著低于仅用CII免疫的小鼠，且前者在研究期间体重增加。此外，喂食CII（250 - 270）的小鼠脾脏细胞中特异性T细胞活性、增殖和干扰素（IFN）-γ分泌受到积极抑制，喂食CII（250 - 270）的小鼠血清抗CII、抗CII（250 - 27）抗体活性以及特异性抗体形成脾脏细胞的频率显著低于仅用CII免疫的小鼠。这些观察结果表明，口服CII（250 - 270）可抑制胶原诱导性关节炎中的细胞免疫和体液免疫反应，并且在单细胞水平同时分析抗原特异性细胞免疫和体液免疫反应将有助于理解CIA中的口服耐受机制以及开发针对RA的创新治疗干预措施。
