Conformation of HIV-1 Envelope governs rhesus CD4 usage and simian-human immunodeficiency virus replication

Abstract

Infection of rhesus macaques with simian-human immunodeficiency viruses (SHIVs) is the preferred model system for vaccine development because SHIVs encode HIV-1 envelope glycoproteins (Env) – a key target of HIV-1 neutralizing antibodies. Since the goal of vaccines is to prevent new infections, SHIVs encoding circulating HIV-1 Env are desired as challenge viruses. Development of such biologically relevant SHIVs has been challenging as they fail to infect rhesus macaques, mainly because most circulating HIV-1 Env do not use rhesus CD4 (rhCD4) receptor for viral entry. Most primary HIV-1 Env exist in a closed conformation and occasionally transit to downstream, open conformation through an obligate intermediate conformation. Here, we provide genetic evidence that open Env conformations can overcome the rhCD4 entry barrier and increase replication of SHIVs in rhesus lymphocytes. Consistent with prior studies, we found that circulating HIV-1 Env do not use physiological levels of rhCD4 efficiently for viral entry. However, using HIV-1 Env with single amino acid substitutions that alter their conformational state, we found that transitions to intermediate and open Env conformation allow usage of rhCD4 for viral entry. To this end, we engineered these single amino acid substitutions in the transmitted/founder HIV-1BG505 Env encoded by SHIV-BG505 and found that open Env conformation enhances SHIV replication in rhesus lymphocytes. Lastly, CD4-mediated SHIV pull-down, sensitivity to soluble CD4, and fusogenicity assays indicated that open Env conformation promotes efficient rhCD4 binding and viral-host membrane fusion. These findings identify conformational state of HIV-1 Env as a major determinant for rhCD4 usage, viral fusion, and SHIV replication.

HIV-1 Envelope Glycoproteins Proteolytic Cleavage Protects Infected Cells from ADCC Mediated by Plasma from Infected Individuals

“…Besides Env-CD4 interaction, there are also structural features of HIV-1 Env that can modulate the sensitivity of HIV-1 to ADCC responses mediated by nnAbs present in plasma from infected individuals. Natural polymorphisms in the Phe43 cavity (notably in CRF01_AE strains) and mutations of conserved residues in the trimer association domain have been shown to modulate Env conformation [25,[56][57][58][59] and as a result, the susceptibility of cells infected with these viruses to ADCC responses [51,60,61]. Similarly, proteolytic cleavage has been reported to stabilize a "closed" Env conformation [62][63][64][65], since mutations in the furin cleavage site resulted in the spontaneous sampling of downstream conformations, including Env State 2A [40,55,63].…”