Microbiology Graduate School Training Program, Department of Microbiology, Mount Sinai School of Medicine, New York, NY, USA.
J Virol. 2012 Jul;86(14):7484-95. doi: 10.1128/JVI.00230-12. Epub 2012 May 2.
Human immunodeficiency virus type 1 (HIV-1) infection can spread efficiently from infected to uninfected T cells through adhesive contacts called virological synapses (VSs). In this process, cell-surface envelope glycoprotein (Env) initiates adhesion and viral transfer into an uninfected recipient cell. Previous studies have found some HIV-1-neutralizing patient sera to be less effective at blocking VS-mediated infection than infection with cell-free virus. Here we employ sensitive flow cytometry-based infection assays to measure the inhibitory potency of HIV-1-neutralizing monoclonal antibodies (MAb) and HIV-1-neutralizing patient sera against cell-free and VS-mediated infection. To various degrees, anti-Env MAbs exhibited significantly higher 50% inhibitory concentration (IC(50)s) against VS-mediated infection than cell-free infection. Notably, the MAb 17b, which binds a CD4-induced (CD4i) epitope on gp120, displayed a 72-fold reduced efficacy against VS-mediated inocula compared to cell-free inocula. A mutant with truncation mutation in the gp41 cytoplasmic tail (CT) which is unable to modulate Env fusogenicity in response to virus particle maturation but which can still engage in cell-to-cell infection was tested for the ability to resist neutralizing antibodies. The ΔCT mutation increased cell surface staining by neutralizing antibodies, significantly enhanced neutralization of VS-mediated infection, and had reduced or no effect on cell-free infection, depending upon the antibody. Our results suggest that the gp41 CT regulates the exposure of key neutralizing epitopes during cell-to-cell infection and plays an important role in immune evasion. Vaccine strategies should consider immunogens that reflect Env conformations exposed on the infected cell surface to enhance protection against VS-mediated HIV-1 spread.
人类免疫缺陷病毒 1 型(HIV-1)感染可以通过称为病毒学突触(VS)的粘附接触从感染的 T 细胞高效传播到未感染的 T 细胞。在这个过程中,细胞表面包膜糖蛋白(Env)启动粘附和病毒转移到未感染的受体细胞中。先前的研究发现,一些 HIV-1 中和患者血清在阻断 VS 介导的感染方面不如无细胞病毒感染有效。在这里,我们采用灵敏的基于流式细胞术的感染测定法来测量 HIV-1 中和单克隆抗体(MAb)和 HIV-1 中和患者血清对无细胞和 VS 介导的感染的抑制效力。在不同程度上,抗 Env MAb 对 VS 介导的感染的 50%抑制浓度(IC 50 )表现出明显高于无细胞感染的抑制活性。值得注意的是,与无细胞接种物相比,结合 gp120 上的 CD4 诱导(CD4i)表位的 MAb 17b 对 VS 介导的接种物的功效降低了 72 倍。测试了不能响应病毒颗粒成熟而调节 Env 融合性但仍能进行细胞间感染的具有 gp41 细胞质尾巴(CT)截断突变的突变体抵抗中和抗体的能力。ΔCT 突变增加了中和抗体的细胞表面染色,显著增强了对 VS 介导的感染的中和作用,并且取决于抗体,对无细胞感染的影响降低或没有。我们的结果表明,gp41 CT 调节细胞间感染过程中关键中和表位的暴露,并在免疫逃避中起重要作用。疫苗策略应考虑反映感染细胞表面上暴露的 Env 构象的免疫原,以增强对 VS 介导的 HIV-1 传播的保护。