Synergistic latency reversal by the SMAC mimetic AZD5582 and iBET JQ1, combined with Nef ablation, facilitates immune-mediated elimination of latently HIV-1-infected T-cells

The shock-and-kill strategy can reactivate latent HIV-1 in vivo but has failed to meaningfully reduce the reactivatable reservoir, suggesting insufficient immunological visibility of infected cells. To investigate latency reversal agent (LRA)-specific shortcomings, we developed a novel HIV-1 latency model - Jurkat E6.1 subclonal T-cell lines, each harbouring a single, full-length, NL4.3-based provirus with a GFPOPT reporter in the Env V5 loop - and quantified HIV-1 reactivation, Env surface presentation across LRAs and synergistically acting combinations. Bryostatin-1 induced GBP5-mediated restriction of surface Env and resistance to apoptosis. Bryostatin-1/JQ1 synergised for HIV-1 reactivation and boosted levels of cell-surface Env, but failed to reduce resistance to apoptosis. AZD5582/JQ1 boosted cell-surface Env levels, without co-induction of GBP5, T-cell activation or apoptotic resistance. HIV-1 Nef antagonised cytotoxic killing of HIV-1-reactivating T-cells - its knockout, combined with AZD5582/JQ1, achieved the highest immune-mediated elimination of latently infected T-cells, representing a promising HIV cure approach.