Hicar Mark D, Chen Xuemin, Kalams Spyros A, Sojar Hakimuddin, Landucci Gary, Forthal Donald N, Spearman Paul, Crowe James E
Department of Pediatrics, University at Buffalo, Buffalo, NY 14222, United States; Department of Microbiology and Immunology, University at Buffalo, Buffalo, NY 14222, United States.
Department of Pediatrics, Emory University School of Medicine, Atlanta, GA 30322, United States; Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322, United States.
Mol Immunol. 2016 Feb;70:94-103. doi: 10.1016/j.molimm.2015.12.002. Epub 2015 Dec 31.
Neutralizing antibodies (Abs) are thought to be a critical component of an appropriate HIV vaccine response. It has been proposed that Abs recognizing conformationally dependent quaternary epitopes on the HIV envelope (Env) trimer may be necessary to neutralize diverse HIV strains. A number of recently described broadly neutralizing monoclonal Abs (mAbs) recognize complex and quaternary epitopes. Generally, many such Abs exhibit extensive numbers of somatic mutations and unique structural characteristics. We sought to characterize the native antibody (Ab) response against circulating HIV focusing on such conformational responses, without a prior selection based on neutralization. Using a capture system based on VLPs incorporating cleaved envelope protein, we identified a selection of B cells that produce quaternary epitope targeting Abs (QtAbs). Similar to a number of broadly neutralizing Abs, the Ab genes encoding these QtAbs showed extensive numbers of somatic mutations. However, when expressed as recombinant molecules, these Abs failed to neutralize virus or mediate ADCVI activity. Molecular analysis showed unusually high numbers of mutations in the Ab heavy chain framework 3 region of the variable genes. The analysis suggests that large numbers of somatic mutations occur in Ab genes encoding HIV Abs in chronically infected individuals in a non-directed, stochastic, manner.
中和抗体(Abs)被认为是适当的HIV疫苗反应的关键组成部分。有人提出,识别HIV包膜(Env)三聚体上构象依赖性四级表位的抗体可能是中和多种HIV毒株所必需的。最近描述的一些广泛中和单克隆抗体(mAbs)识别复杂的四级表位。一般来说,许多此类抗体表现出大量的体细胞突变和独特的结构特征。我们试图表征针对循环HIV的天然抗体(Ab)反应,重点关注此类构象反应,而不进行基于中和的预先筛选。使用基于包含裂解包膜蛋白的病毒样颗粒(VLPs)的捕获系统,我们鉴定出了一系列产生靶向四级表位抗体(QtAbs)的B细胞。与许多广泛中和抗体相似,编码这些QtAbs的抗体基因显示出大量的体细胞突变。然而,当作为重组分子表达时,这些抗体未能中和病毒或介导ADCVI活性。分子分析表明,可变基因的抗体重链框架3区域存在异常大量的突变。分析表明,在慢性感染个体中,编码HIV抗体的抗体基因以非定向、随机的方式发生大量体细胞突变。