Fig. 3: Multi-parameter immunofluorescence (mIF) of the patient-derived samples reveal that increased infiltration of T-cells and response to immunotherapy can be predicted by SA based Act+/Inf+ phenotypes.
From: The responses of HNSCC patients to immunotherapy are shown by two novel co-expression patterns
A CD8 and PD-L1 measurements using mIF in patients 4698 (Act+/Inf+) and 4321 (Act−/Inf−). B Average # of CD8+ T-cells per sample was plotted vs. the amplitude (\({\lambda }_{2}\left(k\right)\)) of Inf+ phenotype (process 2) of each sample k. (Pearson r correlation). C Schematic view of cell-proximity analysis performed using the R “phenoptr” package on the mIF images. CD8+ T-cells within the separation distance of 10, 20, 30, or 40 µm between the T-cells and PD-L1+ cancer cells were counted. D Quantification of CD8+ T-cells as shown in (C) (2-way ANOVA test ****p value < 0.0001). E Comparison of cytokine levels between individuals with activation-phenotype and patients without the phenotype (T-test P value *<0.05, **<0.01, *** <0.001, ****<0.0001). F “Before and after” treatment plots present the levels of cytokines associated with response to immunotherapy. For each patient, fold change of cytokine levels after immunotherapy (IM, anti-PD-1 gray, anti-GITR red, Combination orange) compared to the mean level of untreated explants (ctrl) was calculated. Patient with Act+ phenotype (4905) was compared to the Act+/Inf+ patients (4950 and 4698). (Wilcoxon signed-rank test and Mann–Whitney test were used, p value *<0.05, ***<0.001, ****<0.0001).
