Evaluation of differential G-protein subunit markers for HIV latent infection in Jurkat and A3R5.7 cells
Human immunodeficiency virus (HIV) infects helper T-cells by binding to the CD4 and CCR5/CXCR4 receptors for viral entry. The available antiretroviral therapy (ART) prevents progression towards acquired immunodeficiency syndrome (AIDS), but interruption of ART causes HIV rebound. This rebound is due to latent HIV reservoirs. There is no diagnostic method available to detect latently infected T-cells. CCR5 and CXCR4 are G-protein coupled receptors (GPCRs), which regulate the activation of G-proteins. Our prevous studies have shown that during viral entry, HIV uses CXCR4/CCR5 to activate G-proteins to facilitate infection. We are interseted in knowing whether the establishment of HIV reservoir in T cells can alter T cell G-proteins to facilitate viral persistence. G-proteins are heterotrimers of α, β, and γ subunits, each subunit performs different signaling roles in cells. Our lab developed a Multiplex G-protein profiling system. This system can identify G-protein subunits expressed in lymphocytes. We profiled G-proteins in HIV-infected cells and uninfected cells. Lymphocytes were infected with HIV-1 for five days. p24 expression was then analyzed by intracellular staining using flow cytometry. Lymphocytes showed p24 expression and were prepared for RNA extraction. RNA from cells is then used for our Multiplex G-Protein profiling system. The expression of G-protein subunits was then analyzed by gel-electrophoresis. The identification of the specific G-proteins subunits and the pattern expressed in infected HIV cells may be a powerful tool for detecting HIV latent reservoirs.
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