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目的 制备和鉴定具有鼻咽癌相关抗原内影像的抗独特型单克隆抗体 (Ab2 )。方法 用鼻咽癌单抗 (Ab1)免疫小鼠 ,免疫鼠脾细胞与骨髓瘤细胞SP2 / 0融合获得杂交瘤细胞系。用SandwichELISA和结合抑制试验筛选阳性克隆。为进一步证实Ab2是否具有鼻咽癌相关抗原的内影像 ,用Ab2免疫小鼠获得抗血清 ,以ELISA和竞争抑制试验检测Ab3。用迟发性变态反应和T细胞增殖试验检测Ab2诱导的细胞免疫反应。结果 选用两株抗鼻咽癌单抗FC2、HNL5 (Ab1)为免疫原 ,制备两株抗相应Ab1独特型的抗独特型抗体 2H4和5D3(Ab2 )。 2H4、5D3能分别与FC2、HNL5特异性结合 ,并能抑制相应Ab1与靶细胞HNE2的反应 ,诱导产生能与相应Ab1竞争结合靶抗原的抗抗独特型抗体 (Ab3)。并能诱发特异性迟发型超敏反应 (DTH)和细胞增殖反应。结论 抗独特型抗体 2H4、5D3具有鼻咽癌相关抗原的内影像 (Ab2 β) ,能模拟鼻咽癌相关抗原诱导体液和细胞免疫反应。
Objective To prepare and identify an anti-idiotypic monoclonal antibody (Ab2) with an internal image of nasopharyngeal carcinoma-associated antigen. Methods Mice were immunized with nasopharyngeal carcinoma monoclonal antibody (Ab1) and the murine spleen cells were fused with myeloma cells SP2 / 0 to obtain hybridoma cell lines. Sandwich ELISA and binding inhibition assays were used to screen positive clones. To further confirm whether Ab2 has an internal image of NPC associated antigen, antisera were obtained from mice immunized with Ab2 and Ab3 detected by ELISA and competition inhibition assays. Ab2-induced cellular immune responses were examined using the Delayed Allergy and T-Cell Proliferation Assay. Results Two monoclonal antibodies against nasopharyngeal carcinoma FC2 and HNL5 (Ab1) were used as immunogens to prepare two anti-idiotypic antibodies 2H4 and 5D3 (Ab2). 2H4 and 5D3 can specifically bind to FC2 and HNL5, respectively, and can inhibit the reaction between the corresponding Ab1 and the target cell HNE2 to induce the production of an anti-idiotypic antibody (Ab3) that can compete with the corresponding Ab1 for binding to the target antigen. And can induce specific delayed-type hypersensitivity (DTH) and cell proliferation response. Conclusion Anti-idiotypic antibodies 2H4 and 5D3 have the internal image of nasopharyngeal carcinoma-associated antigen (Ab2 β), which can simulate the nasopharyngeal carcinoma-associated antigen-induced humoral and cellular immune responses.