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Fig. 4 | Journal for ImmunoTherapy of Cancer

Fig. 4

From: Single-cell multiplexed cytokine profiling of CD19 CAR-T cells reveals a diverse landscape of polyfunctional antigen-specific response

Fig. 4

Polyfunctional heat map and PAT PCA reveal distinct CD4+ CAR-T cell profiles across donors. a Polyfunctional heat map displaying major functional subsets secreted across the 4 donors’ CD4+ 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. Donor 1, closely followed by donor 4, has the highest frequencies of most expressed functional groups. Donor 3 is less polyfunctional, while donor 2 has virtually no secreted polyfunctional groups. The group GM-CSF, Granzyme B, IL-13 and TNF-α is expressed exclusively by the CD4+ CARs of donors 1 and 4, but not by the CARs of donor 2 or donor 3. Similarly, the 7-plex group containing GM-CSF, Granzyme B, IFN-γ, IL-8, IL-13, MIP-1α, and TNF-α is unique to these two donors. Functional groups not containing GM-CSF or IL-13 are expressed at similar frequencies by donor 3 as they are by donors 1 and 4. 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. The lack of donor 2 (orange) subsets indicates the lower polyfunctionality of this sample, while the presence of numerous donor 1 (blue) and 4 (green) groups in the right area of the graph indicates the highly-polyfunctional makeup of these two samples. Donor 3 has generally less polyfunctional subsets, typically including combinations of Granzyme B, MIP-1α, IL-8, and TNF-α but lacking IFN-γ, IL-13, and GM-CSF. Donor 4 largely spans the polyfunctional profiles of both donors 1 and 3

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