Fig. 6: Deficiencies of specific B-cell populations in toxicity patients (n = 8) identified in an independent validation NSCLC cohort.

a Heatmap showing normalised expression of the base B-cell panel markers for the 20 B-cell clusters identified with FlowSOM. The cluster IDs and relative frequencies are displayed as a bar graph on the right-hand side, along with the median scaled expression. b UMAP plots stratified according to condition, “no toxicity”, “toxicity” and “healthy controls”. Clusters highlighted by blue boxes correspond to clusters identified from (A). These clusters are attenuated in the “toxicity” cohort in this unsupervised analysis. The box labelled “A” is encircling clusters 16 (mustard) and 17 (lilac). The box labelled “B” is encircling cluster 4 (light blue). The box labelled “C” is encircling cluster 12 (light green). All samples are randomly downsampled to account for equally representative populations across samples. c Diffusion map stratified according to condition, “no toxicity”, “toxicity” and “healthy controls”. Clusters labelled 4, 12, 16, 17 and 20 are attenuated in the “toxicity” cohort in this unsupervised analysis. Cluster 4 is shown by the thin “light blue” line of cells to the right of the darker blue cells in cluster 3. All samples are randomly downsampled to account for equally representative populations across samples. d Bulk expression UMAP stratified according to IL-10 expression, which demonstrates less intense IL-10 expression in the toxicity cohort compared to the non-toxicity patients. Expression is largely confined to the Breg clusters demarcated by the red arrows. Matched IL-10 expression in the corresponding clusters in the toxicity patients is reduced.