A. Tumor infiltrating lymphocytes (TIL) were isolated from a resected melanoma, expanded in the presence of IL-2, and frozen for storage in liquid nitrogen. Thawed TIL were rested for 24 hrs in the absence of any cytokines. Upon stimulation with PMA plus Ionomycin for 6 hours, both CD8+ and CD4+ T cells secreted high levels of the IL-4 and little or no IFN-γ. T cells were tested by ICS. Results are representative of 3 separate experiments.
B. TIL were stimulated as above with PMA plus Ionomycin and lysed. mRNA was extracted and subjected to RT-PCR. cDNA for 49 different T cell products was measured by qPCR, and values were normalized to the quantity of actin mRNA. Values shown represent those factors for which PMA treatment resulted in >5 X induction of specific message.