Enhanced expression of IFN-gamma mRNA in CD4(+) or CD8(+) tumour-infiltrating lymphocytes compared to peripheral lymphocytes in patients with renal cell cancer

The mRNA expression of the cytokines IFN-gamma, IL-10 and TNF-alpha and the proapoptotic factor Fas ligand (FasL) was compared in freshly isolated CD4 (+) and CD8(+) tumour-infiltrating lymphocytes (TIL) and simultaneously obt ained autologous CD4(+) and CD8(+) peripheral blood lymphocytes (PBL) from 20 patients with renal cell carcinomas (RCC). TIL were isolated from mechan ically disaggregated tumour material and PBL from peripheral blood by gradi ent centrifugation. The cells of the interphase were depleted from tumour c ells with antihuman epithelial antigen magnetic beads and then positive sel ection was performed with anti-CD4, or anti-CD8 magnetic beads. In these pu re lymphocyte preparations the constitutive expression of cytokine and Fast mRNAs was determined by using a PCR-assisted mRNA amplification assay. In the CD4(+) TIL from the 20 patients with RCC, levels of mRNAs encoding for IFN-gamma (P less than or equal to 0.001), IL-10 (P less than or equal to 0 .05), and Fast (P less than or equal to 0.001) were significantly higher th an in the autologous CD4(+) PBL. Comparison of CD8(+) TIL and CD8(+) PBL re vealed a significant higher expression of IFN-gamma (P less than or equal t o 0.001), IL-10 (P less than or equal to 0.01) and Fast mRNAs (P less than or equal to 0.001) in the former. However, TNF-alpha mRNA levels were signi ficantly lower in the CD8(+) TIL than in the CD8(+) PBL (P less than or equ al to 0.05), These data reflect a general in vivo activation of RCC infiltr ating lymphocytes in the tumour surrounding, (C) 2000 Cancer Research Campa ign.