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. 2000 Aug 16;83(5):637–641. doi: 10.1054/bjoc.2000.1275

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

U Elsässer-Beile 1, M Rindsfüser 1, T Grussenmeyer 1, W Schultze-Seemann 1, U Wetterauer 1
PMCID: PMC2363516  PMID: 10944605

Abstract

The mRNA expression of the cytokines IFN-γ, IL-10 and TNF-α and the proapoptotic factor Fas ligand (FasL) was compared in freshly isolated CD4+and CD8+tumour-infiltrating lymphocytes (TIL) and simultaneously obtained autologous CD4+and CD8+peripheral blood lymphocytes (PBL) from 20 patients with renal cell carcinomas (RCC). TIL were isolated from mechanically disaggregated tumour material and PBL from peripheral blood by gradient centrifugation. The cells of the interphase were depleted from tumour cells with anti-human epithelial antigen magnetic beads and then positive selection was performed with anti-CD4 or anti-CD8 magnetic beads. In these pure lymphocyte preparations the constitutive expression of cytokine and FasL 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-γ (P≤ 0.001), IL-10 (P≤ 0.05), and FasL (P≤ 0.001) were significantly higher than in the autologous CD4+PBL. Comparison of CD8+TIL and CD8+PBL revealed a significant higher expression of IFN-γ (P≤ 0.001), IL-10 (P≤ 0.01) and FasL mRNAs (P≤ 0.001) in the former. However, TNF-α mRNA levels were significantly lower in the CD8+TIL than in the CD8+PBL (P≤ 0.05). These data reflect a general in vivo activation of RCC infiltrating lymphocytes in the tumour surrounding. © 2000 Cancer Research Campaign

Keywords: renal cell carcinoma, TIL, cytokines, Fas ligand

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Selected References

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  1. Alderson M. R., Tough T. W., Davis-Smith T., Braddy S., Falk B., Schooley K. A., Goodwin R. G., Smith C. A., Ramsdell F., Lynch D. H. Fas ligand mediates activation-induced cell death in human T lymphocytes. J Exp Med. 1995 Jan 1;181(1):71–77. doi: 10.1084/jem.181.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Angevin E., Kremer F., Gaudin C., Hercend T., Triebel F. Analysis of T-cell immune response in renal cell carcinoma: polarization to type 1-like differentiation pattern, clonal T-cell expansion and tumor-specific cytotoxicity. Int J Cancer. 1997 Jul 29;72(3):431–440. doi: 10.1002/(sici)1097-0215(19970729)72:3<431::aid-ijc10>3.0.co;2-f. [DOI] [PubMed] [Google Scholar]
  3. Asselin-Paturel C., Echchakir H., Carayol G., Gay F., Opolon P., Grunenwald D., Chouaib S., Mami-Chouaib F. Quantitative analysis of Th1, Th2 and TGF-beta1 cytokine expression in tumor, TIL and PBL of non-small cell lung cancer patients. Int J Cancer. 1998 Jul 3;77(1):7–12. doi: 10.1002/(sici)1097-0215(19980703)77:1<7::aid-ijc2>3.0.co;2-y. [DOI] [PubMed] [Google Scholar]
  4. Belldegrun A., Muul L. M., Rosenberg S. A. Interleukin 2 expanded tumor-infiltrating lymphocytes in human renal cell cancer: isolation, characterization, and antitumor activity. Cancer Res. 1988 Jan 1;48(1):206–214. [PubMed] [Google Scholar]
  5. Berke G. The CTL's kiss of death. Cell. 1995 Apr 7;81(1):9–12. doi: 10.1016/0092-8674(95)90365-8. [DOI] [PubMed] [Google Scholar]
  6. Brouwenstijn N., Gaugler B., Krüse K. M., van der Spek C. W., Mulder A., Osanto S., van den Eynde B. J., Schrier P. I. Renal-cell carcinoma-specific lysis by cytotoxic T-lymphocyte clones isolated from peripheral blood lymphocytes and tumor-infiltrating lymphocytes. Int J Cancer. 1996 Oct 9;68(2):177–182. doi: 10.1002/(SICI)1097-0215(19961009)68:2<177::AID-IJC6>3.0.CO;2-V. [DOI] [PubMed] [Google Scholar]
  7. Brunner T., Mogil R. J., LaFace D., Yoo N. J., Mahboubi A., Echeverri F., Martin S. J., Force W. R., Lynch D. H., Ware C. F. Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates activation-induced apoptosis in T-cell hybridomas. Nature. 1995 Feb 2;373(6513):441–444. doi: 10.1038/373441a0. [DOI] [PubMed] [Google Scholar]
  8. Caignard A., Guillard M., Gaudin C., Escudier B., Triebel F., Dietrich P. Y. In situ demonstration of renal-cell-carcinoma-specific T-cell clones. Int J Cancer. 1996 May 16;66(4):564–570. doi: 10.1002/(SICI)1097-0215(19960516)66:4<564::AID-IJC23>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  9. Chen L., Linsley P. S., Hellström K. E. Costimulation of T cells for tumor immunity. Immunol Today. 1993 Oct;14(10):483–486. doi: 10.1016/0167-5699(93)90262-J. [DOI] [PubMed] [Google Scholar]
  10. Dhein J., Walczak H., Bäumler C., Debatin K. M., Krammer P. H. Autocrine T-cell suicide mediated by APO-1/(Fas/CD95) Nature. 1995 Feb 2;373(6513):438–441. doi: 10.1038/373438a0. [DOI] [PubMed] [Google Scholar]
  11. Elsässer-Beile U., Grussenmeyer T., Gierschner D., Schmoll B., Schultze-Seemann W., Wetterauer U., Schulte Mönting J. Semiquantitative analysis of Th1 and Th2 cytokine expression in CD3+, CD4+, and CD8+ renal-cell-carcinoma-infiltrating lymphocytes. Cancer Immunol Immunother. 1999 Jul;48(4):204–208. doi: 10.1007/s002620050566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Elsässer-Beile U., Wetterauer U., Schultze-Seemann W., Gallati H., Mönting J. S., von Kleist S. Analysis of the immune reactivity of infiltrating and peripheral lymphocytes from patients with renal cell carcinoma by measuring cytokine secretion. Cancer Immunol Immunother. 1996 Feb;42(2):93–98. doi: 10.1007/s002620050257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Filgueira L., Zuber M., Merlo A., Caetano V., Schultz E., Harder F., Spagnoli G. C., Heberer M. Cytokine gene transcription in renal cell carcinoma. Br J Surg. 1993 Oct;80(10):1322–1325. doi: 10.1002/bjs.1800801034. [DOI] [PubMed] [Google Scholar]
  14. Finke J. H., Rayman P., Edinger M., Tubbs R. R., Stanley J., Klein E., Bukowski R. Characterization of a human renal cell carcinoma specific cytotoxic CD8+ T cell line. J Immunother (1991) 1992 Jan;11(1):1–11. doi: 10.1097/00002371-199201000-00001. [DOI] [PubMed] [Google Scholar]
  15. Finke J. H., Rayman P., Hart L., Alexander J. P., Edinger M. G., Tubbs R. R., Klein E., Tuason L., Bukowski R. M. Characterization of tumor-infiltrating lymphocyte subsets from human renal cell carcinoma: specific reactivity defined by cytotoxicity, interferon-gamma secretion, and proliferation. J Immunother Emphasis Tumor Immunol. 1994 Feb;15(2):91–104. doi: 10.1097/00002371-199402000-00002. [DOI] [PubMed] [Google Scholar]
  16. Igarashi T., Murakami S., Takahashi H., Matsuzaki O., Shimazaki J. Changes on distribution of CD4+/CD45RA- and CD8+/CD11- cells in tumor-infiltrating lymphocytes of renal cell carcinoma associated with tumor progression. Eur Urol. 1992;22(4):323–328. doi: 10.1159/000474780. [DOI] [PubMed] [Google Scholar]
  17. Ito N., Nakamura H., Tanaka Y., Ohgi S. Lung carcinoma: analysis of T helper type 1 and 2 cells and T cytotoxic type 1 and 2 cells by intracellular cytokine detection with flow cytometry. Cancer. 1999 Jun 1;85(11):2359–2367. [PubMed] [Google Scholar]
  18. Kowalczyk D., Skorupski W., Kwias Z., Nowak J. Flow cytometric analysis of tumour-infiltrating lymphocytes in patients with renal cell carcinoma. Br J Urol. 1997 Oct;80(4):543–547. doi: 10.1046/j.1464-410x.1997.00408.x. [DOI] [PubMed] [Google Scholar]
  19. Maeurer M. J., Martin D. M., Castelli C., Elder E., Leder G., Storkus W. J., Lotze M. T. Host immune response in renal cell cancer: interleukin-4 (IL-4) and IL-10 mRNA are frequently detected in freshly collected tumor-infiltrating lymphocytes. Cancer Immunol Immunother. 1995 Aug;41(2):111–121. doi: 10.1007/BF01527407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Martínez-Lorenzo M. J., Alava M. A., Anel A., Piñeiro A., Naval J. Release of preformed Fas ligand in soluble form is the major factor for activation-induced death of Jurkat T cells. Immunology. 1996 Dec;89(4):511–517. doi: 10.1046/j.1365-2567.1996.d01-782.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mitropoulos D., Kooi S., Rodriguez-Villanueva J., Platsoucas C. D. Characterization of fresh (uncultured) tumour-infiltrating lymphocytes (TIL) and TIL-derived T cell lines from patients with renal cell carcinoma. Clin Exp Immunol. 1994 Aug;97(2):321–327. doi: 10.1111/j.1365-2249.1994.tb06088.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nagoshi M., Goedegebuure P. S., Burger U. L., Sadanaga N., Chang M. P., Eberlein T. J. Successful adoptive cellular immunotherapy is dependent on induction of a host immune response triggered by cytokine (IFN-gamma and granulocyte/macrophage colony-stimulating factor) producing donor tumor-infiltrating lymphocytes. J Immunol. 1998 Jan 1;160(1):334–344. [PubMed] [Google Scholar]
  23. Nagoshi M., Sadanaga N., Joo H. G., Goedegebuure P. S., Eberlein T. J. Tumor-specific cytokine release by donor T cells induces an effective host anti-tumor response through recruitment of host naive antigen presenting cells. Int J Cancer. 1999 Jan 18;80(2):308–314. doi: 10.1002/(sici)1097-0215(19990118)80:2<308::aid-ijc22>3.0.co;2-o. [DOI] [PubMed] [Google Scholar]
  24. Nakagomi H., Pisa P., Pisa E. K., Yamamoto Y., Halapi E., Backlin K., Juhlin C., Kiessling R. Lack of interleukin-2 (IL-2) expression and selective expression of IL-10 mRNA in human renal cell carcinoma. Int J Cancer. 1995 Nov 3;63(3):366–371. doi: 10.1002/ijc.2910630311. [DOI] [PubMed] [Google Scholar]
  25. Ng S. Y., Gunning P., Eddy R., Ponte P., Leavitt J., Shows T., Kedes L. Evolution of the functional human beta-actin gene and its multi-pseudogene family: conservation of noncoding regions and chromosomal dispersion of pseudogenes. Mol Cell Biol. 1985 Oct;5(10):2720–2732. doi: 10.1128/mcb.5.10.2720-2732.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Olive C., Nicol D., Falk M. C. Characterisation of gamma delta T cells in renal cell carcinoma patients by polymerase chain reaction analysis of T cell receptor transcripts. Cancer Immunol Immunother. 1997 Mar;44(1):27–34. doi: 10.1007/s002620050351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ortegel J. W., Staren E. D., Faber L. P., Warren W. H., Braun D. P. Cytokine biosynthesis by tumor-infiltrating T lymphocytes from human non-small-cell lung carcinoma. Cancer Immunol Immunother. 2000 Feb;48(11):627–634. doi: 10.1007/s002620050011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pisa P., Halapi E., Pisa E. K., Gerdin E., Hising C., Bucht A., Gerdin B., Kiessling R. Selective expression of interleukin 10, interferon gamma, and granulocyte-macrophage colony-stimulating factor in ovarian cancer biopsies. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7708–7712. doi: 10.1073/pnas.89.16.7708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Raff T., van der Giet M., Endemann D., Wiederholt T., Paul M. Design and testing of beta-actin primers for RT-PCR that do not co-amplify processed pseudogenes. Biotechniques. 1997 Sep;23(3):456–460. doi: 10.2144/97233st02. [DOI] [PubMed] [Google Scholar]
  30. Schendel D. J., Gansbacher B., Oberneder R., Kriegmair M., Hofstetter A., Riethmüller G., Segurado O. G. Tumor-specific lysis of human renal cell carcinomas by tumor-infiltrating lymphocytes. I. HLA-A2-restricted recognition of autologous and allogeneic tumor lines. J Immunol. 1993 Oct 15;151(8):4209–4220. [PubMed] [Google Scholar]
  31. Türeci O., Sahin U., Vollmar E., Siemer S., Göttert E., Seitz G., Parkkila A. K., Shah G. N., Grubb J. H., Pfreundschuh M. Human carbonic anhydrase XII: cDNA cloning, expression, and chromosomal localization of a carbonic anhydrase gene that is overexpressed in some renal cell cancers. Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7608–7613. doi: 10.1073/pnas.95.13.7608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Van den Hove L. E., Van Gool S. W., Van Poppel H., Baert L., Coorevits L., Van Damme B., Ceuppens J. L. Phenotype, cytokine production and cytolytic capacity of fresh (uncultured) tumour-infiltrating T lymphocytes in human renal cell carcinoma. Clin Exp Immunol. 1997 Sep;109(3):501–509. doi: 10.1046/j.1365-2249.1997.4771375.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wang Q., Redovan C., Tubbs R., Olencki T., Klein E., Kudoh S., Finke J., Bukowski R. M. Selective cytokine gene expression in renal cell carcinoma tumor cells and tumor-infiltrating lymphocytes. Int J Cancer. 1995 Jun 9;61(6):780–785. doi: 10.1002/ijc.2910610607. [DOI] [PubMed] [Google Scholar]
  34. Whiteside T. L., Miescher S., Hurlimann J., Moretta L., von Fliedner V. Separation, phenotyping and limiting dilution analysis of T-lymphocytes infiltrating human solid tumors. Int J Cancer. 1986 Jun 15;37(6):803–811. doi: 10.1002/ijc.2910370602. [DOI] [PubMed] [Google Scholar]
  35. Whiteside T. L., Parmiani G. Tumor-infiltrating lymphocytes: their phenotype, functions and clinical use. Cancer Immunol Immunother. 1994 Jul;39(1):15–21. doi: 10.1007/BF01517175. [DOI] [PMC free article] [PubMed] [Google Scholar]

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