Fresh or thawed PBMCs were immunophenotyped for T cell subsets and reported as percentages of total PBMCs and as percentages of total T cells in the case of T cell subtypes. Specifically, PBMCs were stained with well-characterized antibodies against markers of interest, including CD3, CD4 plus CD8 according to standard protocols. Results were analyzed using a BD Fortessa multi-color flow cytometer and FlowJo 7.6.1 program (Ashland, OR). Further, immunophenotypic changes of major innate and adaptive immune cells of ICI were evaluated using a minimum of a million PMBCs collected before and after cancer therapy for individual lung cancer patients by a 24-color antibody panel using the “Aurora” Spectral cytometer (Cytek Biosciences, CA). The data analysis and statistical evaluations of this complex data set were performed using the Cytobank (Cytobank Inc., CA) [27].
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