Meyerhoff R Ryan, Scearce Richard M, Ogburn Damon F, Lockwood Brad, Pickeral Joy, Kuraoka Masa, Anasti Kara, Eudailey Joshua, Eaton Amanda, Cooper Melissa, Wiehe Kevin, Montefiori David C, Tomaras Georgia, Ferrari Guido, Alam S Munir, Liao Hua-Xin, Korber Bette, Gao Feng, Haynes Barton F
1 Duke Human Vaccine Institute, Duke University School of Medicine , Durham, North Carolina.
2 Department of Immunology, Duke University School of Medicine , Durham, North Carolina.
AIDS Res Hum Retroviruses. 2017 Aug;33(8):859-868. doi: 10.1089/AID.2016.0294. Epub 2017 May 30.
Antibodies that cross-react with multiple HIV-1 envelopes (Envs) are useful reagents for characterizing Env proteins and for soluble Env capture and purification assays. We previously reported 10 murine monoclonal antibodies induced by group M consensus Env, CON-6 immunization. Each demonstrated broad cross-reactivity to recombinant Envs. Here we characterized the Env epitopes to which they bind. Seven mapped to linear epitopes in gp120, five at the Env N-terminus, and two at the Env C-terminus. One antibody, 13D7, bound at the gp120 N-terminus (aa 30-42), reacted with HIV-1-infected CD4 T cells, and when expressed in a human IgG1 backbone, mediated antibody-dependent cellular cytotoxicity. Antibody 18F11 bound at the gp120 C-terminus (aa 445-459) and reactivity was glycan dependent. Antibodies 13D7, 3B3, and 16H3 bound to 100 percent of HIV-1 Envs tested in ELISA and sodium dodecyl sulfate/polyacrylamide gel electrophoresis/western blot analysis. These data define the epitopes of monoclonal antibody reagents for characterization of recombinant Envs, one epitope of which is also expressed on the surface of HIV-1-infected CD4 T cells.
与多种HIV-1包膜(Env)发生交叉反应的抗体是用于表征Env蛋白以及进行可溶性Env捕获和纯化分析的有用试剂。我们之前报道了由M组共识Env即CON-6免疫诱导产生的10种鼠单克隆抗体。每种抗体都对重组Env表现出广泛的交叉反应性。在此我们对它们所结合的Env表位进行了表征。七种抗体定位到gp120中的线性表位,其中五种位于Env N端,两种位于Env C端。一种抗体13D7在gp120 N端(氨基酸30 - 42)结合,与HIV-1感染的CD4 T细胞发生反应,当在人IgG1骨架中表达时,介导抗体依赖性细胞毒性。抗体18F11在gp120 C端(氨基酸445 - 459)结合,其反应性依赖于聚糖。在酶联免疫吸附测定(ELISA)以及十二烷基硫酸钠/聚丙烯酰胺凝胶电泳/蛋白质免疫印迹分析(SDS/PAGE/Western blot)中,抗体13D7、3B3和16H3与所检测的100%的HIV-1 Env发生结合。这些数据确定了用于表征重组Env的单克隆抗体试剂的表位,其中一个表位也在HIV-1感染的CD4 T细胞表面表达。