抗乙肝病毒核心抗体抗独特型抗体的研究——Ⅱ.制备抗人HBcAb的抗独特型抗体的探讨

The Study of Anti-Idiotype to Anti-[ (Anti-HBc)Id] Ⅱ. Murine Anti-Idiotypic Antibody to Human Anti-HBc

为了研究抗独特型抗体对乙型肝炎免疫反应的调节作用,我们用人的多克隆抗HBc作为免疫原,制备单克隆抗独特型抗体(抗-Id)。将人HBcAb用亲和层析纯化,并经SPA-Sepharose4B柱提取Fab片段。用4株抗正常人IgG的单克隆抗体与人HBcAb复合,制成免疫复合物,再用作免疫原,采用3种不同的免疫方案,除了采用国内外公认的抗原-抗体结合抑制试验检测外,还做了中和试验。在“独特型结合试验”中,在用人HBcAb包放的同时,还用正常人IgG作对照,以排除其抗异种免疫球蛋白的干扰。同时,还进行了交叉反应独特型的筛选工作。我们共对6300个克隆进行了检测,没有检出特异性的抗独特型抗体,却有1450个抗正常人IgG的阳性克隆。对出现上述结果的可能原因进行了探讨。

For the purpose of studying the modulation of anti-idiotypic antibody to immunologic reaction of hepatitis B, BALB/c mice were immunized with human poly-clonal anti-HBc-to prepare monoclonal anti-Id. Three different immunization protocols and three different treated immunogens were used to prepare the hybridomas so as to inhibit the production of anti-human IgG. Human HBcAb were purified by affinity chromatography and Fab fragments were extracted with SPA-Sepharose 4B;human HBcAb componded with a pool of anti-human IgG monoclonal antibodies depressed the response of immunized mice to human IgG; mice were given an intraperitoneal injection of 2.5 mg human IgG 14 days before by the standard method of immunization. The binding test and inhibition test were used to screen anti-idiotypic antibodies. 6 300 hybridoma clonels were screened, however, no specific anti-idiotypic antibo-dy was found, but 1450 positive clonels of anti-human IgG were detected. Different treated immunogens induced different positive clonal rates of anti-human IgG. Fab fragment purified with SPA-Sepharose 48 might cause the lowest proportion of anti-human IgG, both tolerogen and Fab fragment purified with gradient elution had a higher positive clonal rate of anti-human IgG. The reasons for these results were discussed in this report.