摘要
目的制备针对C34和C46单抗, 并以此为工具研究gp41表位,进一步 了解HIV-1包膜蛋白的作用机理和病毒的感染机制,为寻找新的治疗靶点提供抗体工具。 方法常规动物免疫、细胞融合、克隆化制备抗C34与C46的单克隆抗体,并鉴定其 特异性位点，还用ELISA法、MTT法及荧光分子探针技术对单抗的生物学活性进行研究。结果获得了3株抗C34单克隆抗体、2株抗C46单抗。此5个单抗均与C34结合 , 并抑制N36肽与C34肽复合物的形成；其中效价最高的1G1对H9/HIVⅢB细胞的生长有刺 激作用 ,对H9/HIVⅢB细胞和MT-2细胞的融合无明显影响。结论得到5株 可与C34反应,并可抑制C34与N36多肽结合的单抗, 其中1G1所识别的表位可能为增强性或刺激性表位。
Objective To generate monoclonal antibodies against C34， a polypeptide derived from HIV-1 gp41 core structure, and to characterize the epitope of gp41 recognized by McAb. Methods McAb clones against C34 and C46 were prepared by hybridoma technique and identified for epitopes by ELISA, for effect on cell proliferation by MTT, and for effect on cell fusion by molecular fluorescence probe. Results The three of anti-C34 McAb(1G1, 2F8, 2B7) and two of anti-C46 inhibited the binding of N46 and C34. The 1G1, an anti-C34 McAb with the highest titer, can enhance the proliferation of H9/HIV-1ⅢB cells , but can not influence the form of syncytium conformation of HIV-1 in-fected cells(H9/HIV-1ⅢB) with uninfected cells(MT-2). Conclusion These results suggested that the epitope recognized by 1 G1 may be an enhancive epitope which can stimulate the proliferation of HIV-1 infected cell.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2002年第2期89-91,共3页
Immunological Journal
基金
国家自然科学B类基金资助项目(30028021)
