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目的：探讨Ｈｅｙｍａｎｎ肾炎（ＨＮ）致病原受体相关蛋白（ＲＡＰ）羧基端１０８个氨基酸多肽链上是否存在致肾炎的抗原决定簇。方法：应用ＮｅｓｔｅｄＰＣＲ技术，扩增了编码ＲＡＰ羧基端１０８个氨基酸多肽链的ｃＤＮＡ。应用基因重组及亚克隆技术，高效表达了特异性的ＲＡＰ融合蛋白（ＲＡＰ－ｆｐ），并制备了针对该ＲＡＰ－ｆｐ的多抗血清。结果：该多抗血清体外免疫荧光定位于肾小管刷状缘。体内结合试验的直接免疫荧光和免疫电镜表明，在肾小球足突细胞和肾小球基膜之间，形成了典型的被动型Ｈｅｙｍａｎｎ肾炎（ＰＨＮ）免疫复合物。结论：本实验说明天然羧基端ＲＡＰ在体内能被循环抗体结合，在原位形成免疫复合物。首次证明了在ＲＡＰ羧基端１０８个氨基酸多肽链上至少存在一个病理性抗原决定簇。
OBJECTIVE: To investigate the presence or absence of nephritis epitopes on the carboxy-terminal 108 amino acid peptide of Heymann nephritis (HN) -related pathogenic receptor-associated protein (RAP). Methods: NestedPCR technique was used to amplify the cDNA encoding the carboxy terminal 108 amino acids of RAP. Specific RAP fusion protein (RAP-fp) was highly expressed by gene recombination and subcloning techniques and multi-antiserum against this RAP-fp was prepared. Results: The multi-antiserum immunofluorescence in vitro located in the brush border of renal tubules. Direct immunofluorescence and immunoelectron microscopy in vivo binding assays have shown that a typical passive Heymann nephritis (PHN) immune complex is formed between glomerular and green glomerular basement membranes. CONCLUSIONS: This experiment demonstrates that natural carboxy-terminal RAPs can be bound by circulating antibodies in vivo to form immune complexes in situ. It was first demonstrated that there is at least one pathogenic epitope on the 108 amino acid polypeptide chain of RAP.