AAV-Mediated CAR-T therapy eradicate HIV reservoirs with augmented safety and persistence

HIV persists in latent reservoirs within CD4 cells, posing a major barrier to cure. Chimeric antigen receptor (CAR) T cell therapy has shown promise against HIV but is limited by several key challenges, including persistent and protecting CAR T cells expression, particularly those expressing CD4-based receptors from HIV infection, preventing immune evasion through viral mutation, and effectively targeting transcriptionally silent reservoirs that lack antigen expression and thus remain undetectable to CAR-based therapies. AAV based CAR T therapy addresses these challenges by providing stable and safe CAR delivery with low integration risk, enhancing persistence while reducing insertional mutagenesis concerns. AAVCD4CAR transduced primary human T cells at high efficiency and viability; the resulting CAR T cells selectively recognized and killed HIV1 infected CD4+ T cells, significantly reducing viral p24 levels in infected PBMC cultures. In a humanized mouse model engrafted with HIV, AAVCD4CAR T cells led to sustained CAR expression on circulating T cells. Treated mice showed marked suppression of plasma viremia and preservation of CD4+ T cell counts compared to controls, without evidence of systemic toxicity. The CAR T cells exhibited effector cytokine production and memory markers, indicating functional activation and persistence. Moreover, we found that the AAV based CAR is safe and biocompatible and does not affect the physiological function of mice challenged with HIV. Taken together, these results verify that AAVCD4CAR is effective in clearing viral load from ex vivo human samples and in mice models and is safe and biocompatible. Our findings provide proof of concept that an AAVCD4CAR approach can induce durable anti-HIV immunity, suggesting a new avenue for curative therapy.