Directly Infected Resting CD4+T Cells Can Produce HIV Gag without Spreading Infection in a Model of HIV Latency

Abstract

Despite the effectiveness of highly active antiretroviral therapy (HAART) in treating individuals infected with HIV, HAART is not a cure. A latent reservoir, composed mainly of resting CD4+T cells, drives viral rebound once therapy is stopped. Understanding the formation and maintenance of latently infected cells could provide clues to eradicating this reservoir. However, there have been discrepancies regarding the susceptibility of resting cells to HIV infection in vitro and in vivo. As we have previously shown that resting CD4+T cells are susceptible to HIV integration, we asked whether these cells were capable of producing viral proteins and if so, why resting cells were incapable of supporting productive infection. To answer this question, we spinoculated resting CD4+T cells with or without prior stimulation, and measured integration, transcription, and translation of viral proteins. We found that resting cells were capable of producing HIV Gag without supporting spreading infection. This block corresponded with low HIV envelope levels both at the level of protein and RNA and was not an artifact of spinoculation. The defect was reversed upon stimulation with IL-7 or CD3/28 beads. Thus, a population of latent cells can produce viral proteins without resulting in spreading infection. These results have implications for therapies targeting the latent reservoir and suggest that some latent cells could be cleared by a robust immune response.

Author Summary

While HIV is a treatable disease due to effective antiviral therapies, these drugs do not cure HIV. When therapy is stopped, a pool of infected, long-lived, treatment resistant cells re-establishes infection. These latently infected cells, mainly resting CD4+T cells, are barriers to a cure. Studying and understanding the properties of these cells is therefore important to eradicating HIV. It is believed that these latent cells do not produce viral proteins and thus are invisible to the immune system. Here, we show using an in vitro HIV model that a population of latently infected cells can produce HIV Gag. Interestingly, this protein production does not result in the release of detectable infectious virus and so the latent cells are unaffected by antiviral therapy. We therefore examined why some latent cells can produce viral proteins without viral spread. We found that resting cells have the ability to make some of the components required for spreading infection but not all are in sufficient quantity. These results have important implications for treating the latent reservoir, as our work suggests that latent cells might be recognized by a boosted immune response.