Abstract
This study was undertaken to investigate the pathways involved in the interleukin 2 (IL-2)-driven growth of tumour-infiltrating lymphocytes (TILs). For this purpose, TIL lines and freshly isolated TILs obtained from 16 patients with solid cancer (three melanoma, seven primary colorectal carcinoma, four hepatic metastases from colorectal cancer and two lung cancer) were evaluated for (a) expression of IL-2 receptor (IL-2R) both at the RNA level and on the cell surface by flow cytometric analysis and (b) their proliferative activity in response to IL-2 and the role of IL-2R subunits in the IL-2-driven TIL growth. Northern blot analysis showed that TILs express a strong message for both the p55 and the p75 IL-2R. Accordingly, flow cytometric analysis demonstrated that TILs bear both IL-2R chains. TILs cultured in vitro in the presence of rIL-2 were able to proliferate in response to different concentrations of this cytokine. Monoclonal antibodies (MAbs) specifically recognising the p55 and p75 IL-2R chains (anti-Tac and TU27 respectively) exhibited a marked inhibitory effect on IL-2-driven growth when added individually or in appropriate combinations. Our results demonstrated that TILs are equipped with a fully functional IL-2 receptor system, thus suggesting the involvement of this structure in the activation and expansion of TILs following immunotherapy with IL-2.
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- Arima N., Kamio M., Imada K., Hori T., Hattori T., Tsudo M., Okuma M., Uchiyama T. Pseudo-high affinity interleukin 2 (IL-2) receptor lacks the third component that is essential for functional IL-2 binding and signaling. J Exp Med. 1992 Nov 1;176(5):1265–1272. doi: 10.1084/jem.176.5.1265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Balch C. M., Riley L. B., Bae Y. J., Salmeron M. A., Platsoucas C. D., von Eschenbach A., Itoh K. Patterns of human tumor-infiltrating lymphocytes in 120 human cancers. Arch Surg. 1990 Feb;125(2):200–205. doi: 10.1001/archsurg.1990.01410140078012. [DOI] [PubMed] [Google Scholar]
- Belldegrun A., Kasid A., Uppenkamp M., Topalian S. L., Rosenberg S. A. Human tumor infiltrating lymphocytes. Analysis of lymphokine mRNA expression and relevance to cancer immunotherapy. J Immunol. 1989 Jun 15;142(12):4520–4526. [PubMed] [Google Scholar]
- Hatakeyama M., Mori H., Doi T., Taniguchi T. A restricted cytoplasmic region of IL-2 receptor beta chain is essential for growth signal transduction but not for ligand binding and internalization. Cell. 1989 Dec 1;59(5):837–845. doi: 10.1016/0092-8674(89)90607-7. [DOI] [PubMed] [Google Scholar]
- Hatakeyama M., Tsudo M., Minamoto S., Kono T., Doi T., Miyata T., Miyasaka M., Taniguchi T. Interleukin-2 receptor beta chain gene: generation of three receptor forms by cloned human alpha and beta chain cDNA's. Science. 1989 May 5;244(4904):551–556. doi: 10.1126/science.2785715. [DOI] [PubMed] [Google Scholar]
- Ioannides C. G., Fisk B., Tomasovic B., Pandita R., Aggarwal B. B., Freedman R. S. Induction of interleukin-2 receptor by tumor necrosis factor alpha on cultured ovarian tumor-associated lymphocytes. Cancer Immunol Immunother. 1992;35(2):83–91. doi: 10.1007/BF01741854. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Itoh K., Tilden A. B., Balch C. M. Interleukin 2 activation of cytotoxic T-lymphocytes infiltrating into human metastatic melanomas. Cancer Res. 1986 Jun;46(6):3011–3017. [PubMed] [Google Scholar]
- Kawakami Y., Rosenberg S. A., Lotze M. T. Interleukin 4 promotes the growth of tumor-infiltrating lymphocytes cytotoxic for human autologous melanoma. J Exp Med. 1988 Dec 1;168(6):2183–2191. doi: 10.1084/jem.168.6.2183. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kim T. Y., von Eschenbach A. C., Filaccio M. D., Hayakawa K., Parkinson D. R., Balch C. M., Itoh K. Clonal analysis of lymphocytes from tumor, peripheral blood, and nontumorous kidney in primary renal cell carcinoma. Cancer Res. 1990 Sep 1;50(17):5263–5268. [PubMed] [Google Scholar]
- Kradin R. L., Kurnick J. T., Lazarus D. S., Preffer F. I., Dubinett S. M., Pinto C. E., Gifford J., Davidson E., Grove B., Callahan R. J. Tumour-infiltrating lymphocytes and interleukin-2 in treatment of advanced cancer. Lancet. 1989 Mar 18;1(8638):577–580. doi: 10.1016/s0140-6736(89)91609-7. [DOI] [PubMed] [Google Scholar]
- Minamoto S., Mori H., Hatakeyama M., Kono T., Doi T., Ide T., Uede T., Taniguchi T. Characterization of the heterodimeric complex of human IL-2 receptor alpha.beta chains reconstituted in a mouse fibroblast cell line, L929. J Immunol. 1990 Oct 1;145(7):2177–2182. [PubMed] [Google Scholar]
- Muul L. M., Spiess P. J., Director E. P., Rosenberg S. A. Identification of specific cytolytic immune responses against autologous tumor in humans bearing malignant melanoma. J Immunol. 1987 Feb 1;138(3):989–995. [PubMed] [Google Scholar]
- Noguchi M., Adelstein S., Cao X., Leonard W. J. Characterization of the human interleukin-2 receptor gamma chain gene. J Biol Chem. 1993 Jun 25;268(18):13601–13608. [PubMed] [Google Scholar]
- Pandolfi F., Boyle L. A., Trentin L., Kurnick J. T., Isselbacher K. J., Gattoni-Celli S. Expression of HLA-A2 antigen in human melanoma cell lines and its role in T-cell recognition. Cancer Res. 1991 Jun 15;51(12):3164–3170. [PubMed] [Google Scholar]
- Robb R. J., Greene W. C. Internalization of interleukin 2 is mediated by the beta chain of the high-affinity interleukin 2 receptor. J Exp Med. 1987 Apr 1;165(4):1201–1206. doi: 10.1084/jem.165.4.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg S. A. Immunotherapy and gene therapy of cancer. Cancer Res. 1991 Sep 15;51(18 Suppl):5074s–5079s. [PubMed] [Google Scholar]
- Rosenberg S. A., Packard B. S., Aebersold P. M., Solomon D., Topalian S. L., Toy S. T., Simon P., Lotze M. T., Yang J. C., Seipp C. A. Use of tumor-infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. A preliminary report. N Engl J Med. 1988 Dec 22;319(25):1676–1680. doi: 10.1056/NEJM198812223192527. [DOI] [PubMed] [Google Scholar]
- Schwartzentruber D. J., Topalian S. L., Mancini M., Rosenberg S. A. Specific release of granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, and IFN-gamma by human tumor-infiltrating lymphocytes after autologous tumor stimulation. J Immunol. 1991 May 15;146(10):3674–3681. [PubMed] [Google Scholar]
- Shimizu Y., Iwatsuki S., Herberman R. B., Whiteside T. L. Effects of cytokines on in vitro growth of tumor-infiltrating lymphocytes obtained from human primary and metastatic liver tumors. Cancer Immunol Immunother. 1991;32(5):280–288. doi: 10.1007/BF01789045. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith K. A. Interleukin-2: inception, impact, and implications. Science. 1988 May 27;240(4856):1169–1176. doi: 10.1126/science.3131876. [DOI] [PubMed] [Google Scholar]
- Takeshita T., Goto Y., Tada K., Nagata K., Asao H., Sugamura K. Monoclonal antibody defining a molecule possibly identical to the p75 subunit of interleukin 2 receptor. J Exp Med. 1989 Apr 1;169(4):1323–1332. doi: 10.1084/jem.169.4.1323. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takeshita T., Ohtani K., Asao H., Kumaki S., Nakamura M., Sugamura K. An associated molecule, p64, with IL-2 receptor beta chain. Its possible involvement in the formation of the functional intermediate-affinity IL-2 receptor complex. J Immunol. 1992 Apr 1;148(7):2154–2158. [PubMed] [Google Scholar]
- Topalian S. L., Solomon D., Rosenberg S. A. Tumor-specific cytolysis by lymphocytes infiltrating human melanomas. J Immunol. 1989 May 15;142(10):3714–3725. [PubMed] [Google Scholar]
- Tsudo M., Karasuyama H., Kitamura F., Tanaka T., Kubo S., Yamamura Y., Tamatani T., Hatakeyama M., Taniguchi T., Miyasaka M. The IL-2 receptor beta-chain (p70). Ligand binding ability of the cDNA-encoding membrane and secreted forms. J Immunol. 1990 Jul 15;145(2):599–606. [PubMed] [Google Scholar]
- Tsudo M., Kitamura F., Miyasaka M. Characterization of the interleukin 2 receptor beta chain using three distinct monoclonal antibodies. Proc Natl Acad Sci U S A. 1989 Mar;86(6):1982–1986. doi: 10.1073/pnas.86.6.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uchiyama T., Broder S., Waldmann T. A. A monoclonal antibody (anti-Tac) reactive with activated and functionally mature human T cells. I. Production of anti-Tac monoclonal antibody and distribution of Tac (+) cells. J Immunol. 1981 Apr;126(4):1393–1397. [PubMed] [Google Scholar]
- Vaccarello L., Wang Y. L., Whiteside T. L. Sustained outgrowth of autotumor-reactive T lymphocytes from human ovarian carcinomas in the presence of tumor necrosis factor alpha and interleukin 2. Hum Immunol. 1990 Jun;28(2):216–227. doi: 10.1016/0198-8859(90)90022-h. [DOI] [PubMed] [Google Scholar]
- Voss S. D., Leary T. P., Sondel P. M., Robb R. J. Identification of a direct interaction between interleukin 2 and the p64 interleukin 2 receptor gamma chain. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2428–2432. doi: 10.1073/pnas.90.6.2428. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Voss S. D., Robb R. J., Weil-Hillman G., Hank J. A., Sugamura K., Tsudo M., Sondel P. M. Increased expression of the interleukin 2 (IL-2) receptor beta chain (p70) on CD56+ natural killer cells after in vivo IL-2 therapy: p70 expression does not alone predict the level of intermediate affinity IL-2 binding. J Exp Med. 1990 Oct 1;172(4):1101–1114. doi: 10.1084/jem.172.4.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Waldmann T. A. The interleukin-2 receptor. J Biol Chem. 1991 Feb 15;266(5):2681–2684. [PubMed] [Google Scholar]
- Wang H. M., Smith K. A. The interleukin 2 receptor. Functional consequences of its bimolecular structure. J Exp Med. 1987 Oct 1;166(4):1055–1069. doi: 10.1084/jem.166.4.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wang Y. L., Si L. S., Kanbour A., Herberman R. B., Whiteside T. L. Lymphocytes infiltrating human ovarian tumors: synergy between tumor necrosis factor alpha and interleukin 2 in the generation of CD8+ effectors from tumor-infiltrating lymphocytes. Cancer Res. 1989 Nov 1;49(21):5979–5985. [PubMed] [Google Scholar]
- Yagita M., Itoh K., Tsudo M., Schaub L. B., Platsoucas C. D., Balch C. M., Grimm E. A. Involvement of both Tac and non-Tac interleukin 2-binding peptides in the interleukin 2-dependent proliferation of human tumor-infiltrating lymphocytes. Cancer Res. 1989 Mar 1;49(5):1154–1159. [PubMed] [Google Scholar]
- Zambello R., Trentin L., Pizzolo G., Bulian P., Masciarelli M., Feruglio C., Agostini C., Raimondi R., Chisesi T., Semenzato G. Cell membrane expression and functional role of the p75 subunit of interleukin-2 receptor in lymphoproliferative disease of granular lymphocytes. Blood. 1990 Nov 15;76(10):2080–2085. [PubMed] [Google Scholar]

