Monoclonal anti-idiotype antibody bearing the internal image of nasopharyngeal carcinoma associated antigen

具有鼻咽癌相关抗原内影像的抗独特型单克隆抗体研究(英文)

Objective To generate and characterize anti-idiotypic monoclonal antibody (Ab2) that bears the internal image of nasopharyngeal carcinoma (NPC) associated antigen. Methods Using NPC monoclonal antibody (Ab1) as immunogen, hybridoma cells were obtained by fusion of SP2/0 myeloma cells with immunized murine spleen cells. Positive clones were screened by Sandwich ELISA and a binding inhibition test. To determine whether Ab2 possess the internal image of the original antigen or not, mice were immunized with Ab2. ELISA and the competitive inhibition assay tested anti-anti-idiotypic antibodies (Ab3) in anti-sera. Cell-mediated immunity to tumors induced by Ab2 was investigated by a delayed-type hypersensitivity response and the mouse T-cell proliferation assay. Results Anti-idiotypic monoclonal antibodies against the monoclonal anti-NPC antibodies FC2 and HNL5 were generated that recognize NPC associated antigens. These Ab2, which were designated 2H4 and 5D3, could inhibit the binding of FC2 or HNL5 to NPC cell lines. Anti-sera from the immunized mice, which contained Ab3, could compete with FC2 or HNL5 for binding with NPC cell by a competitive inhibition assay. Mice immunized with 2H4 or 5D3 coupled with keyhole limpet hemocyanin (KLH), showed a positive and specific delayed-type hypersensitivity (DTH) reaction after stimulation by NPC cells. The mouse T cell proliferative assay indicated that there was a significantly higher proliferative response of the splenocytes in the experimental groups than that in control groups. Conclusions Anti-idiotypic antibodies 2H4 and 5D3 are Ab2 beta bearing the internal image of the epitope of NPC associated antigen. Either 2H4 or 5D3 expressing three-dimensional shapes that resemble the structure of natural antigens could induce humoral and cellular immune response.

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。