Myers L K, Cooper S W, Terato K, Seyer J M, Stuart J M, Kang A H
Department of Pediatrics, University of Tennessee, Memphis.
Clin Immunol Immunopathol. 1995 Apr;75(1):33-8. doi: 10.1006/clin.1995.1049.
The murine model of collagen-induced arthritis is characterized by the development of an immune response against joint cartilage. Arthritis can be significantly suppressed by the administration of type II collagen (CII) or one of the CNBr peptides, CB11 (CII 124-402) as a tolerogen prior to immunization. We have previously shown that two synthetic peptides, representing sequences CII 260-270 and CII 181-209, are effective tolerogens. In this paper, we now characterize the T cell determinant with CII 181-209. A series of synthetic peptides overlapping CII 181-209 and analogs of chick CII 181-209 containing site-directed amino acid substitutions was developed and cultured with T cells from DBA/1 mice immunized with CII. Supernatants were collected and analyzed for the presence of the T cell lymphokine IFN-gamma. These data indicate the critical T cell determinant to be located within CII 190-200. This conclusion is further supported by the observation that an unodecapeptide representing CII 190-200 was just as effective as CII 181-209 in suppressing arthritis and anti-CII antibody response when tested as a tolerogen. Analogs containing single amino acid substitutions at residues 191, 194, 197, 198, or 200 were significantly less effective in inducing T cell responses. Each of these peptide analogs was then given as neonatal tolerogens to DBA/1 mice. Mice were subsequently immunized and observed for the development of arthritis. These studies identified residues 194, 197, 198, and 200, and probably residue 191, as critical for tolerance and the suppression of arthritis. Elucidation of the fine structures of T cell determinants which are critical for suppression of arthritis should allow these techniques to be used for developing specific immunotherapeutic approaches to autoimmune arthritis.
胶原诱导性关节炎的小鼠模型的特征是针对关节软骨产生免疫反应。在免疫前给予II型胶原（CII）或作为耐受原的CNBr肽之一CB11（CII 124 - 402），可显著抑制关节炎。我们之前已经表明，代表序列CII 260 - 270和CII 181 - 209的两种合成肽是有效的耐受原。在本文中，我们现在对CII 181 - 209的T细胞决定簇进行了表征。开发了一系列与CII 181 - 209重叠的合成肽以及含有定点氨基酸取代的鸡CII 181 - 209类似物，并与用CII免疫的DBA/1小鼠的T细胞一起培养。收集上清液并分析T细胞淋巴因子IFN-γ的存在情况。这些数据表明关键的T细胞决定簇位于CII 190 - 200内。当作为耐受原进行测试时，代表CII 190 - 200的十一肽在抑制关节炎和抗CII抗体反应方面与CII 181 - 209同样有效，这一观察结果进一步支持了这一结论。在残基191、194、197、198或200处含有单个氨基酸取代的类似物在诱导T细胞反应方面明显效果较差。然后将这些肽类似物中的每一种作为新生耐受原给予DBA/1小鼠。随后对小鼠进行免疫并观察关节炎的发展情况。这些研究确定了残基194、197、198和200，可能还有残基191，对于耐受和关节炎的抑制至关重要。阐明对关节炎抑制至关重要的T细胞决定簇的精细结构，应该能够使这些技术用于开发针对自身免疫性关节炎的特异性免疫治疗方法。
