In vivo killing of primary HIV-infected cells by peripheral-injected early memory–enriched anti-HIV duoCAR T cells
Kim Anthony-Gonda, … , Harris Goldstein, Boro Dropulić
Kim Anthony-Gonda, … , Harris Goldstein, Boro Dropulić
Published November 8, 2022
Citation Information: JCI Insight. 2022;7(21):e161698. https://doi.org/10.1172/jci.insight.161698.
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Research Article AIDS/HIV Therapeutics

In vivo killing of primary HIV-infected cells by peripheral-injected early memory–enriched anti-HIV duoCAR T cells

Abstract

HIV-specific chimeric antigen receptor–T cell (CAR T cell) therapies are candidates to functionally cure HIV infection in people with HIV (PWH) by eliminating reactivated HIV-infected cells derived from latently infected cells within the HIV reservoir. Paramount to translating such therapeutic candidates successfully into the clinic will require anti-HIV CAR T cells to localize to lymphoid tissues in the body and eliminate reactivated HIV-infected cells such as CD4+ T cells and monocytes/macrophages. Here we show that i.v. injected anti-HIV duoCAR T cells, generated using a clinical-grade anti-HIV duoCAR lentiviral vector, localized to the site of active HIV infection in the spleen of humanized mice and eliminated HIV-infected PBMCs. CyTOF analysis of preinfusion duoCAR T cells revealed an early memory phenotype composed predominantly of CCR7+ stem cell–like/central memory T cells (TSCM/TCM) with expression of some effector-like molecules. In addition, we show that anti-HIV duoCAR T cells effectively sense and kill HIV-infected CD4+ T cells and monocytes/macrophages. Furthermore, we demonstrate efficient genetic modification of T cells from PWH on suppressive ART into anti-HIV duoCAR T cells that subsequently kill autologous PBMCs superinfected with HIV. These studies support the safety and efficacy of anti-HIV duoCAR T cell therapy in our presently open phase I/IIa clinical trial (NCT04648046).

Authors

Kim Anthony-Gonda, Alex Ray, Hang Su, Yuge Wang, Ying Xiong, Danica Lee, Ariele Block, Vanessa Chilunda, Jessica Weiselberg, Lily Zemelko, Yen Y. Wang, Sarah Kleinsorge-Block, Jane S. Reese, Marcos de Lima, Christina Ochsenbauer, John C. Kappes, Dimiter S. Dimitrov, Rimas Orentas, Steven G. Deeks, Rachel L. Rutishauser, Joan W. Berman, Harris Goldstein, Boro Dropulić

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Figure 4

Anti-HIV duoCAR T cells recognize and potently kill HIV-infected monocytes.

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Anti-HIV duoCAR T cells recognize and potently kill HIV-infected monocyt...
(A–C) Purified and matured monocytes (A), PBMCs (B), or CD4+ T cells (C) from the same donor were infected with HIVBaL-LucR virus for 2 days (monocytes) or 3 days (PBMCs and CD4+ T cells). HIV-infected cells were either untreated (infected) or treated with donor-matched untransduced (UTD) T cells or MND-ΔW duoCAR T cells (duoCAR) at an E:T ratio of 1:1 for an additional 3 days. Uninfected cells were used as negative controls in the assay. The magnitude of HIV-1 infection was quantified 3 days after infection by measuring Renilla luciferase activity and was expressed as relative light units (RLU). Data are shown as mean ± SEM. The percent HIV-1 suppression is shown above the bar graph for MND-ΔW duoCAR T cells and was calculated relative to infected cells either left untreated or treated with UTD control T cells. The study shown is from 6 independent experiments with cells from 6 different HIV-1 seronegative donors. Statistical analysis was performed by unpaired Student’s t test. Significance is considered P < 0.05. (D) Sensitivity of anti-HIV duoCAR T cell killing against monocytes, PBMCs, and CD4+ T cells. Percent killing of HIV-infected cells was calculated relative to UTD control T cells. Data are shown as mean ± SEM. Data show n = 4 HIV seronegative donors for all E:T ratios except 1:100, for which data show n = 3. Statistical analysis was performed by 2-way ANOVA followed by Tukey’s multiple comparison post hoc test.