Guyver Carly J, Copland David A, Calder Claudia J, Sette Alessandro, Sidney John, Dick Andrew D, Nicholson Lindsay B
Department of Cellular and Molecular Medicine, School of Medical Sciences, University of Bristol, United Kingdom.
Invest Ophthalmol Vis Sci. 2006 May;47(5):2027-35. doi: 10.1167/iovs.05-0984.
Experimental autoimmune uveoretinitis (EAU) can be induced in C57BL/6 mice (I-A(b)) using human retinoid-binding protein-3 (hRBP-3, previously IRBP) residues 1-20. This study of a truncated murine peptide (mRBP-3 1-16) was conducted to determine its pathogenic potential and to characterize partially its interaction with specific T cells.
After immunization with mRBP-3 1-16 or hRBP-3 1-20, EAU was assessed by immunohistochemistry. The immune response was assessed by tritiated thymidine incorporation and cytokine production analyzed by enzyme-linked immunosorbent assay (ELISA). T-cell receptor (TCR)- and major histocompatibility complex (MHC)-binding of mRBP-3 1-16 was studied by modeling and by using altered peptide ligands (APLs) and T-cell clones.
mRBP-3 1-16 induced EAU in C57BL/6 mice, with severity and kinetics comparable to that after immunization with hRBP-3 1-20. T cells taken from mice immunized with mRBP-3 1-16 had a Th1 phenotype and proliferated in response to reactivation with mRBP-3 1-16, hRBP-3 1-20, or mRBP-3 1-16 APLs. mRBP-3 1-16 APLs elicited at least five distinct patterns of reactivity when tested with the mRBP-3 1-16-reactive T-cell clones.
mRBP-3 1-16 immunizes and causes EAU in C57BL/6 mice. The studies using T-cell clones and APLs demonstrate that the immune response to mRBP-3 1-16 is drawn from a diverse population of antigen-specific T cells with a Th1 phenotype. Modeling and analysis of clones indicate that nonpathogenic T cells of an mRBP-3 1-16-reactive T-cell line recognize the peptide in a single register.
使用人视黄醇结合蛋白-3（hRBP-3，以前称为IRBP）的1-20位残基可在C57BL/6小鼠（I-A(b)）中诱导实验性自身免疫性葡萄膜视网膜炎（EAU）。本研究对截短的鼠肽（mRBP-3 1-16）进行了检测，以确定其致病潜力，并部分表征其与特异性T细胞的相互作用。
用mRBP-3 1-16或hRBP-3 1-20免疫后，通过免疫组织化学评估EAU。通过氚标记胸腺嘧啶核苷掺入评估免疫反应，并通过酶联免疫吸附测定（ELISA）分析细胞因子产生。通过建模以及使用改变的肽配体（APL）和T细胞克隆研究mRBP-3 1-16与T细胞受体（TCR）和主要组织相容性复合体（MHC）的结合。
mRBP-3 1-16在C57BL/6小鼠中诱导出EAU，其严重程度和动力学与用hRBP-3 1-20免疫后的情况相当。从用mRBP-3 1-16免疫的小鼠中获取的T细胞具有Th1表型，并在再次用mRBP-3 1-16、hRBP-3 1-20或mRBP-3 1-16 APL激活后增殖。当用mRBP-3 1-16反应性T细胞克隆进行测试时，mRBP-3 1-16 APL引发了至少五种不同的反应模式。
mRBP-3 1-16免疫并在C57BL/6小鼠中引起EAU。使用T细胞克隆和APL的研究表明，对mRBP-3 1-16的免疫反应来自具有Th1表型的多种抗原特异性T细胞群体。对克隆的建模和分析表明，mRBP-3 1-16反应性T细胞系的非致病性T细胞以单一排列方式识别该肽。
