Fig. 1 |. BLT mouse-derived HIV-specific CAR T cells are functionally indistinguishable from human-derived CAR T cells in vitro.

αCD3/CD28 Dynabeads were used to activate purified human T cells from a BLT mouse and healthy human donor, and then cells were transduced with the CD4-based CAR.ζ construct coexpressing GFP. a, FACS plots identify CAR.ζ T cells from each T cell source as GFP⁺ and CD4⁺. b, Following 10 d of culture, CD8⁺ CAR.ζ T cells were mixed with HIV_YU2 Envelope⁺ K562 cells (K.Env) and upregulation of human cytokines was measured. c, Polyfunctionality profiles for combinatorial subsets of CD4⁺ and CD8⁺ CAR.ζ T cells producing 0 to 5 of the human cytokines GM-CSF, IFN-γ, IL-2, MIP-1β and TNF. Average of three unique donors per T cell source. d,e, HIV suppression assay as described in the Methods. d, FACS plots indicating the frequency of HIV-infected cells 6 d after coculturing with BLT mouse- or human-derived CAR.ζ T cells at denoted E/T ratios. e, Summary of frequency of HIV-infected cells (live CAR⁻ CD8⁻ cells) at 2, 4 and 6 d after coculture with BLT mouse- or human-derived CAR.ζ and untransduced (UTD) T cells at indicated E/T ratios. f,g, HIV elimination assay as described in the Methods. FACS plots (f) and summarized data (g) for frequency of active caspase-3 within HIV-infected cells (live CTV⁺ HIV_GAG⁺ cells) 24 h after coculture with BLT mouse- or human-derived CAR.ζ and UTD T cells at 1:1 E/T ratio. Each symbol represents the average of technical duplicates per donor (n = 3 biologically independent donors). e,g, Symbols and lines reflect mean and error bars indicate ±s.e.m. RD1 is another name for phycoerythrin (PE).