Figure 1.
NHP model of CAR T-cell therapy reveals CD8+ CARneg T cells with an activation signature. (A) Schematic of sample preparation: T cells, CARpos, and CARneg cells were flow cytometrically sorted before infusion, from the product, at day 10 (peak expansion) and day 14 (the start of CAR T-cell contraction), and prepared for scRNA-seq and scTCR-seq. (B) CAR T-cell expansion and B-cell aplasia were tracked in animal R.315. (C) Uniform manifold approximation and projection (UMAP) with Leiden clustering of the scRNA-seq data set, colored by the time point. (D) UMAP of the scRNA-seq data set colored by flow cytometrically sorted CARpos and CARneg cells. (E) UMAP of the scRNA-seq data set, colored by the normalized CAR transcript counts. (F) Unadjusted CAR transcript counts in the sorted CARpos and CARneg populations. (G) decoupleR weighted sum (WSUM) analysis of a naïve vs memory gene signature in CD4+ CARpos and CARneg T cells and a naïve vs effector gene signature in CD8+ CARpos and CARneg T cells, with the analysis performed with cells isolated at the time of peak expansion. (H) Violin plots of normalized expression of the CD8+ effector molecules granzyme B (GZMB), IFN-γ, and perforin 1 (PRF1) in CD4+ and CD8+ CARpos and CARneg T cells.

NHP model of CAR T-cell therapy reveals CD8+ CARneg T cells with an activation signature. (A) Schematic of sample preparation: T cells, CARpos, and CARneg cells were flow cytometrically sorted before infusion, from the product, at day 10 (peak expansion) and day 14 (the start of CAR T-cell contraction), and prepared for scRNA-seq and scTCR-seq. (B) CAR T-cell expansion and B-cell aplasia were tracked in animal R.315. (C) Uniform manifold approximation and projection (UMAP) with Leiden clustering of the scRNA-seq data set, colored by the time point. (D) UMAP of the scRNA-seq data set colored by flow cytometrically sorted CARpos and CARneg cells. (E) UMAP of the scRNA-seq data set, colored by the normalized CAR transcript counts. (F) Unadjusted CAR transcript counts in the sorted CARpos and CARneg populations. (G) decoupleR weighted sum (WSUM) analysis of a naïve vs memory gene signature in CD4+ CARpos and CARneg T cells and a naïve vs effector gene signature in CD8+ CARpos and CARneg T cells, with the analysis performed with cells isolated at the time of peak expansion. (H) Violin plots of normalized expression of the CD8+ effector molecules granzyme B (GZMB), IFN-γ, and perforin 1 (PRF1) in CD4+ and CD8+ CARpos and CARneg T cells.

or Create an Account

Close Modal
Close Modal