HIV-1gp41上ELDKWA中和表位及单克隆抗体的研究

发布时间：2018-01-01 02:00

本文关键词：HIV-1gp41上ELDKWA中和表位及单克隆抗体的研究　出处：《清华大学》2009年博士论文　论文类型：学位论文

更多相关文章： HIV gp41 单克隆抗体 ELDKWA表位

【摘要】： 位于人免疫缺陷病毒(HIV-1)跨膜蛋白gp41上的ELDKWA表位是重要中和表位,与之结合的人源单克隆抗体2F5具有广泛的中和活性。然而,国际期刊论文多次报导,通过人工构建含有该表位的免疫原,所诱导出表位特异性的单克隆抗体,均未表现出与2F5相同的中和活性。为了揭示其原因,本论文工作围绕ELDKWA表位及针对该表位的单克隆抗体,继续深入研究和探讨了单克隆抗体具有的特性。本研究以gp41上的重要中和表位ELDKWA为靶位点,设计表位多肽抗原并制备出单克隆抗体。对本实验室原有的和重新制备的单克隆抗体的研究发现:(1)实验所用免疫原的设计影响抗体应答的效果,即使是相同表位多肽抗原,诱导出的单克隆抗体与重组表达的gp41的结合能力也存在显著差异;(2)在膜融合实验中,这些单抗各自表现出不同的抑制活性;(3)体外病毒中和实验(前期工作)和假病毒中和实验表明,不同的单抗在中和能力方面存在巨大差异,能够中和HIV-1 B亚型野毒株92US657的单抗,不能中和通过转染人工构建的B亚型假病毒株JRFL,同时,具有假病毒中和活性的单抗,对于野毒株92US657也不具有中和效果。这些研究结果提示,针对同一表位的单抗,其与抗原的结合能力、抑制膜融合的能力以及中和能力和中和活性也可能受到病毒株本身序列及抗体空间结构的影响。
[Abstract]:The ELDKWA epitope located on the transmembrane protein gp41 of human immunodeficiency virus (HIV-1) is an important neutralization epitope, with which the human monoclonal antibody 2F5 has extensive neutralization activity. It has been reported many times in international journals that the epitope specific monoclonal antibody induced by artificial construction of the immunogen containing the epitope does not exhibit the same neutralization activity as 2F5. In this paper, the characteristics of ELDKWA epitopes and monoclonal antibodies against them have been studied. In this study, ELDKWA, an important neutralizing epitope on gp41, was used as the target. The epitope peptide antigen was designed and monoclonal antibody was prepared. It was found that the design of immunogen used in the experiment affected the effect of antibody response. Even with the same epitope polypeptide antigen, the binding ability of the induced monoclonal antibody to the recombinant gp41 was significantly different. (2) in the membrane fusion experiment, these McAbs showed different inhibitory activities. In vitro neutralization experiments (previous work) and pseudoviral neutralization experiments showed that there were significant differences in neutralization ability of different McAbs. The McAb of HIV-1 B subtype wild strain 92US657 could not neutralize the constructed virus strain JRFL, and the McAb with pseudovirus-neutralizing activity could not be neutralized. These results suggest that the monoclonal antibody against the same epitope has the ability to bind to the antigen. The ability of inhibiting membrane fusion, neutralization ability and neutralization activity may also be affected by the sequence of virus strain and the space structure of antibody.
【学位授予单位】：清华大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392

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