Specht Grijp-Glandorf R J, De Heer E, Dekker-Nooren C C, Daha M R, Van Es L A
Department of Nephrology, University Hospital Leiden, The Netherlands.
Immunology. 1990 Jul;70(3):290-5.
The aim of this study was to characterize the epitope specificity and the idiotype(s) of gp330-specific autoantibodies in active Heymann nephritis (HN). To study this, two monoclonal autoantibodies (mAb), 3D9B and 5B8C, derived from Lewis rats with HN, were generated. These antibodies bound to different epitopes on the autoantigen gp330, as shown by in vitro studies. In order to characterize the idiotype(s) of the mAb, rabbit anti-idiotypic antisera were prepared. It was shown that the rabbit antisera directed against the 3D9B as well as the 5B8C idiotype were able to inhibit the binding of nephritic eluate to gp330. This indicated that the idiotypes recognized on 3D9B and 5B8C were relevant to the spectrum of idiotypes involved in HN. In order to investigate the nature of the two idiotypes of these mAb, cross-absorption and binding studies were performed. The rabbit antisera raised against one of the two idiotypes bound in a direct binding ELISA to both idiotypes. Since the observed cross-reactivity of the antisera suggested that the idiotypes of 3D9B and 5B8C were at least partly cross-reactive, homologous polyclonal and monoclonal anti-idiotypic antibodies were prepared by immunizing rats with 3D9B or 5B8C. The results from the binding and inhibition experiments strongly suggest that the idiotypes of 3D9B and 5B8C contain cross-reactive or public idiotopes.
本研究的目的是鉴定活动性海曼肾炎(HN)中gp330特异性自身抗体的表位特异性和独特型。为了研究这一点,从患有HN的Lewis大鼠中制备了两种单克隆自身抗体(mAb),即3D9B和5B8C。体外研究表明,这些抗体与自身抗原gp330上的不同表位结合。为了鉴定mAb的独特型,制备了兔抗独特型抗血清。结果显示,针对3D9B以及5B8C独特型的兔抗血清能够抑制肾炎洗脱液与gp330的结合。这表明在3D9B和5B8C上识别的独特型与HN中涉及的独特型谱相关。为了研究这些mAb的两种独特型的性质,进行了交叉吸收和结合研究。在直接结合ELISA中,针对两种独特型之一产生的兔抗血清与两种独特型均结合。由于观察到的抗血清交叉反应性表明3D9B和5B8C的独特型至少部分交叉反应,因此通过用3D9B或5B8C免疫大鼠制备了同源多克隆和单克隆抗独特型抗体。结合和抑制实验的结果强烈表明,3D9B和5B8C的独特型含有交叉反应性或公共独特型表位。