Expansion of tumor-T cell pairs from fine needle aspirates of melanoma metastases

Lymphocytes expanded fi om excised specimens can be used to characterize in tratumoral T cell responses. These analyses, however, are limited to one ti me point in the natural history of the removed tumor. The expansion of auto logous tumor cells and tumor-infiltrating lymphocytes (TIL) from fine needl e aspirates (FNA) of tumors potentially allows a dynamic evaluation of T ce ll responses within the same lesion at moments relevant to the disease cour se or response to therapy. Fourteen TIL cultures and 8 tumor cell lines cr ere generated from 18 FNA (12 patients). Five of six Tn that could be teste d against autologous tumor demonstrated specific reactivity. Two additional TIL for which no autologous tumor was available demonstrated recognition o f HLA-matched melanoma cell lines. Serial FNA of the same lesions were perf ormed in five HLA-A*0201 patients vaccinated with the emulsified melanoma A g (MA) epitopes: MART-1:27-35; tyrosinase:368-376(370D); gp100:280-288(288V ); and gp100:209-217 (210M). FNA material was separately cultured for a sho rt time in IL-2 (300 IU/ml) after stimulation with irradiated autologous PB MC pulsed with each peptide or FluM1:58-66 (1 mu mol/ml). No peptide-specif ic TIL could be expanded from prevaccination FN,I, However, after vaccinati on, TIL specific for gp100:280(g280), gp100:209 (g209), and MART-1:27-35 (M ART-1)-related epitopes were identified in three, three, and two patients, respectively, No Flu reactivity could be elicited in TIL, whereas it was co nsistently present in Dal allel PBMC cultures, This excluded PBMC contamina tion of the FNA material. This analysis suggests the feasibility of TIL exp ansion from minimal FNA material and localization of vaccine-specific T cel ls at the tumor site.