To investigate whether NSCLC cells could directly impact T cell differentiation, CD4+ T cells from peripheral blood mononuclear cells (PBMCs) of healthy donors were pre-incubated with human NSCLC cell line A549 cells for 12 h, activated with anti-CD3/CD28 beads for 4 days, and detected for T cell differentiations by quantifying lineage-determining transcriptional factors and cytokines with flow cytometry (Fig. S1A). This evidence concerns the gene CD28 and non-small cell lung carcinoma.