NK cells engineered with a chimeric antigen receptor delay HIV rebound and reshape HIV reservoir composition

Abstract Durable HIV remission requires strategies that eliminate or permanently silence the latent reservoir that persists despite antiretroviral therapy (ART). We engineer allogeneic human primary natural killer (NK) cells expressing a truncated CD4-based chimeric antigen receptor (D1D2 CAR) targeting the conserved CD4 binding site on HIV Env. D1D2 CAR NK cells selectively kill HIV-infected primary CD4 T cells in vitro and significantly delay viral rebound following ART interruption in humanized mice infected with barcoded CCR5-tropic HIV. Barcoded HIV tracking shows reductions in the number and diversity of rebounding viral RNA lineages, together with decreased inter-organ sharing of rebound-associated proviral barcodes and integration sites. Integration site and chromatin analyses suggest that rebound-associated proviruses remaining after D1D2 CAR-NK treatment are less frequently located in genic and transcriptionally active chromatin contexts. Thus, CAR-NK cells may target reactivating HIV-infected cells during early ART interruption and shape the composition of the reservoir contributing to rebound.