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