Knospe V, Donoso L A, Banga J P, Yue S, Kasp E, Gregerson D S
Department of Ophthalmology, University of Minnesota, Minneapolis.
Curr Eye Res. 1988 Nov;7(11):1137-47. doi: 10.3109/02713688809001885.
We examined the binding of seven murine monoclonal antibodies raised to S-antigen, an immunopathogenic, 404 residue photoreceptor cell autoantigen which induces experimental autoimmune uveoretinitis. S-antigen has also been identified as arrestin, a protein involved in the regulation of phototransduction. One additional monoclonal antibody (C10C10), raised to a synthetic peptide (peptide N) corresponding to residues 281 to 302 in bovine S-antigen, was also studied. In preliminary studies we examined the specificity of the antibody response to bovine S-antigen in sera from Balb/c mice. Western blots of mouse sera on the cyanogen bromide digest of bovine S-antigen demonstrated that all animals produced antibody which recognized epitopes within the C-terminal cyanogen bromide peptide designated CB46. Mice of the H-2d haplotype, including the Balb/c strain often used to produce monoclonal antibodies, showed little activity to cyanogen bromide peptides other than CB46. Also, all seven of the monoclonals raised to S-antigen are specific for epitopes in the CB46 peptide. The epitopes recognized by the monoclonal antibodies could be grouped into four distinct sites defined by peptides AE-1 (A2G5), peptide AA (PDS-1), peptide 19-OV (A9C6), and peptide 199 (BDS-1,2,3 and 4). The mono-clonal antibody, C10C10, raised to peptide N recognizes an epitope in the N peptide and binds to a larger cyanogen bromide peptide designated CB123 as well as intact S-antigen. Fine mapping of these epitopes was done with various subpeptides. None of the antibodies bound the known immunopathogenic peptide, peptide M, which resides in CB123 although the C10C10 antibody binds a peptide adjacent to peptide M.
我们检测了针对S抗原产生的七种鼠单克隆抗体的结合情况。S抗原是一种具有免疫致病性的、由404个氨基酸残基组成的光感受器细胞自身抗原，可诱发实验性自身免疫性葡萄膜视网膜炎。S抗原也被鉴定为抑制蛋白，一种参与光转导调节的蛋白质。还研究了另一种针对与牛S抗原中281至302位氨基酸残基相对应的合成肽（肽N）产生的单克隆抗体（C10C10）。在初步研究中，我们检测了Balb/c小鼠血清中针对牛S抗原的抗体反应的特异性。用牛S抗原的溴化氰消化物对小鼠血清进行的蛋白质免疫印迹分析表明，所有动物均产生了识别指定为CB46的C端溴化氰肽内表位的抗体。H-2d单倍型的小鼠，包括常用于产生单克隆抗体的Balb/c品系，对除CB46以外的溴化氰肽几乎没有反应。此外，针对S抗原产生的所有七种单克隆抗体均对CB46肽中的表位具有特异性。单克隆抗体识别的表位可分为由肽AE-1（A2G5）、肽AA（PDS-1）、肽19-OV（A9C6）和肽199（BDS-1、2、3和4）定义的四个不同位点。针对肽N产生的单克隆抗体C10C10识别N肽中的一个表位，并与一个更大的指定为CB123的溴化氰肽以及完整的S抗原结合。用各种亚肽对这些表位进行了精细定位。尽管C10C10抗体与肽M相邻的一个肽结合，但没有一种抗体与存在于CB123中的已知免疫致病肽肽M结合。
