Extended Data Fig. 6: T-Cell immunophenotype of CirAE patients Analyzed by Flow Cytometry.
(a) Left: uniform Manifold Approximation and Projection (UMAP) visualization of flow cytometry characterization of CD3+ T-cells from 9 patients at peak of expansion, stratified by CirAE. Middle: CAR percentage and median fluorescence intensity (MFI) in CD3+ T-cells at peak expansion, stratified by CirAE. Violin plots show the mirrored probability density of the data; dashed line indicates the median, and dotted lines denote the 25th and 75th percentiles. Right: proportions of CD4, CD8, and double-negative CART subsets at peak expansion, stratified by CirAE. Bar graphs show the median and the error bars denote the 25th and 75th percentiles. Violin plots show the mirrored probability density of the data; dashed line indicates the median, and dotted lines denote the 25th and 75th percentiles. Statistical comparisons between groups were performed using the two-tailed Wilcoxin test. (b) Flow cytometry analysis of peripheral blood mononuclear cells from 10 patients during CirAE and 6 non-CirAE patients at matched timepoints. CAR+ percentage (top) and MFI (bottom) of CART during CirAE in CD3+ (left), CD3+CD4+ (middle), CD3+CD8+ (right) T-cells. The top rightmost panel depicts absolute CART count (%CAR+ x ALC), stratified by CirAE. Violin plots show the mirrored probability density of the data; dashed line indicates the median, and dotted lines denote the 25th and 75th percentiles. Statistical comparisons between groups were performed using the two-tailed Wilcoxin test. (c) Comparison of peripheral blood (PB) CD4⁺ T-cell proportions between CirAE patients at the time of CirAE and time-matched non-CirAE controls. Left: unpaired comparison of CD4⁺ T-cell proportions at pre-lymphodepletion and CirAE/matched timepoints, stratified by CirAE; bar graphs show the median and the error bars denote the 25th and 75th percentiles. Right: comparison of CD4⁺ T-cell proportions at pre-lymphodepletion and CirAE/matched timepoints in paired longitudinal samples, stratified by CirAE. Statistical comparisons between groups were performed using the two-tailed paired-sample T-test. (d) Flow cytometry immunophenotyping of T-cells from matched PB and cerebrospinal fluid (CSF) samples during Cilta-cel#4’s delayed ICANS episode.
