Fig. 5

Polyfunctional heat map and PAT PCA reveal distinct CD8+ CAR-T cell profiles across donors. a Polyfunctional heat map displaying major functional subsets secreted across the 4 donors’ CD8+ CAR-T samples. Hierarchical clustering is applied to attain a condensed set of functional groups that still faithfully represent the overall profile of the donors. The color-coding indicates how commonly each donor secrets the corresponding functional group/cluster. Donors 1 and 4 are significantly more polyfunctional than donors 2 and 3, and also uniquely secrete the 7-plex group containing GM-CSF, Granzyme B, IFN-γ, IL-8, IL-13, MIP-1α, and TNF-α. Donor 1 has a higher number of unique polyfunctional groups than donor 4, particularly groups not containing IL-8. The only polyfunctional groups secreted by all four donors contain Granzyme B, MIP-1 α with smaller amounts of IFN- γ. b PAT PCA visualization of the same dataset. Data points are color-coded based on donor. Those representing the same functional group are randomly offset, but remain within a radius proportional to the secretion frequency of the corresponding group (i.e., large groups = large circles, small groups = small circles). The principal components are labeled according to their correlation with specific cytokines. Like the CD4+ CAR-T samples, donor 2 (orange) has low polyfunctionality, donors 1 (blue) and 4 (green) are highly polyfunctional. Donor 3 has generally less polyfunctional subsets often comprised of combinations of Granzyme B, MIP-1α, IL-8, and TNF-α but lacks IFN-γ, and IL-13. Donor 4 largely spans the polyfunctional profiles of both donors 1 and 3, which can also be seen in the heat map