Skip to main content
SpringerLink
Log in

Effects of cytokines on in vitro growth of tumor-infiltrating lymphocytes obtained from human primary and metastatic liver tumors

  • Original articles
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Summary

Tumor-infiltrating lymphocytes (TIL) were isolated from 22 human primary and metastatic liver tumors, and expanded in vitro in the presence of either interleukin-2 (IL-2, 100 U/ml) plus tumor necrosis factorα (TNFα, 1000 U/ml), IL-2 (1000 U/ml) plus IL-4 (1000 U/ml) or IL-2 (1000 U/ml) alone. TIL proliferated in culture in 20/22 cases. Among different cytokine combinations, TNFα and IL-2 were most effective in promoting the outgrowth of CD3+ CD8+ T lymphocytes (mean ± SEM: 90%±5) in the cultures of TIL from primary liver tumors. Cytotoxicity against autologous tumor cells was demonstrated in all early cultures of TIL from primary liver cancers in the presence of IL-2 plus TNFα. In contrast, cultures of TIL derived from colon cancer metastatic to liver had significantly lower levels of autotumor cytotoxicity and proportions of CD3+ CD8+ cells (40%±13) than those of TIL from primary liver tumors. The addition on day 0 of interferons (α or γ) to TIL cultured in the presence of TNFα and IL-2, significantly augmented cytotoxicity against autologous tumor. In contrast, incubation of TIL in the presence of IL-4 and IL-2 did not result in increased autotumor responses in the cultures of TIL from primary liver tumors. The expansion (-fold) of TIL (day 30) cultured in the presence of IL-2 alone compared to that in the presence of TNFα and IL-2 was significantly greater for hepatocellular carcinoma (median, 280 vs 260) than for autologous peripheral blood lymphocytes (36 vs 27), cholangiocarcinoma (42 vs 51) or TIL from metastatic colon cancer (39 vs 30). Outgrowth of TIL in IL-2 plus TNFα offers an opportunity for in vitro enrichment in cells with autotumor cytotoxicity in primary liver tumors. However, this cytokine combination was unable to promote and sustain growth of autotumor effectors from TIL in metastatic liver cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Anzano MA, Rieman D, Prichett W, Bowen-Pope DF, Greig R (1989) Growth factor production by human colon carcinoma cell lines. Cancer Res 49: 2898–2904

    Google Scholar 

  2. Cameron RB, McIntosh JK, Rosenberg SA (1988) Synergistic antitumor effects of combination immunotherapy with recombinant interleukin-2 and a recombinant hybrid α-interferon in the treatment of established murine hepatic metastases. Cancer Res 48: 5810–5817

    Google Scholar 

  3. Chen LK, Mathieu-Mahul D, Bach FH, Dausset J, Bensussan A, Sasportes M (1986) Recombinant interferon α can induce rearrangement of T-cell antigen receptor α-chain genes and maturation to cytotoxicity in T-lymphocyte clones in vitro. Proc Natl Acad Sci USA 83: 4887–4889

    Google Scholar 

  4. Chen LK, Tourvieille B, Burns GF, Bach FH, Mathieu-Mahul D, Sasportes M, Bensussan A (1986) Interferon: a cytotoxic T lymphocyte differentiation signal. Eur J Immunol 16: 767–770

    Google Scholar 

  5. Giovarelli M, Santoni A, Jemma C, Musso T, Giuffrida AM, Cavallo G, Landolfo S, Forni G (1988) Obligatory role of IFNγ in induction of lymphokine-activated and T lymphocyte killer activity, but not in boosting of natural cytotoxicity. J Immunol 141: 2831–2836

    Google Scholar 

  6. Gromo G, Geller RL, Inverardi L, Bach FH (1987) Signal requirements in the step-wise functional maturation of cytotoxic T lymphocytes. Nature 327: 424–426

    Google Scholar 

  7. Itoh K, Platsoucas CD, Balch CM (1988) Autologous tumor-specific cytotoxic T lymphocytes in the infiltrate of human metastatic melanomas. Activation by interleukin 2 and autologous tumor cells, and involvement of the T cell receptor. J Exp Med 168: 1419–1441

    Google Scholar 

  8. Kawakami Y, Rosenberg SA, Lotze MT (1988) Interleukin 4 promotes the growth of tumor-infiltrating lymphocytes cytotoxic for human autologous melanoma. J Exp Med 168: 2183–2191

    Google Scholar 

  9. Landolfo S, Cofano F, Giovarelli M, Prat M, Cavallo G, Forni G (1985) Inhibition of interferon-gamma may suppress allograft reactivity by T lymphocytes in vitro and in vivo. Science 229: 176–179

    Google Scholar 

  10. McIntosh JK, Mulé JJ, Krosnick JA, Rosenberg SA (1989) Combination cytokine immunotherapy with tumor necrosis factorα, interleukin 2, and α-interferon and its synergistic antitumor effects in mice. Cancer Res 49: 1408–1414

    Google Scholar 

  11. Miescher S, Stoeck M, Qiao L, Barras C, Barrelet L, Von Fliedner V (1988) Preferential clonogenic deficit of CD8-positive T-lymphocytes infiltrating human solid tumors. Cancer Res 48: 6992–6998

    Google Scholar 

  12. Mulé JJ, Schwarz SL, Roberts AB, Sporn MB, Rosenberg SA (1988) Transforming growth factor-beta inhibits the in vitro generation of lymphokine-activated killer cells and cytotoxic T cells. Cancer Immunol Immunother 26: 95–100

    Google Scholar 

  13. Pross HF, Baines MT, Rubin P, Shragge P, Patterson MS (1981) Spontaneous human lymphocyte mediated cytotoxicity against tumor target cells. IX. The quantitation of natural killer cell activity. J Clin Immunol 1: 51–63

    Google Scholar 

  14. Ranges GE, Figari IS, Espevik T, Palladino MA (1987) Inhibition of cytotoxic T cell development by transforming growth factorβ and reversal by recombinant tumor necrosis factorα. J Exp Med 166: 991–998

    Google Scholar 

  15. Rosenberg SA, Spiess P, Lafreniere R (1986) A new approach to the adoptive immunotherapy of cancer with tumor-infiltrating lymphocytes. Science 233: 1318–1321

    Google Scholar 

  16. Rosenberg SA, Schwarz SL, Spiess PJ (1988) Combination immunotherapy for cancer: Synergistic antitumor interactions of interleukin-2, alpha interferon, and tumor-infiltrating lymphocytes. J Natl Cancer Inst 80: 1393–1397

    Google Scholar 

  17. Shimizu Y, Iwatsuki S, Herberman RB, Whiteside TL (1990) Clonal analysis of tumor-infiltrating lymphocytes from human primary and metastatic liver tumor. Int J Cancer (in press)

  18. Siegel JP (1988) Effects of interferon-γ on the activation of human T lymphocytes. Cell Immunol 111: 461–472

    Google Scholar 

  19. Simon MM, Hochgeschwender U, Brugger U, Landolfo S (1986) Monoclonal antibodies to interferon-γ inhibit interleukin 2-dependent induction of growth and maturation in lectin/antigen-reactive cytolytic T lymphocyte precursors. J Immunol 136: 2755–2762

    Google Scholar 

  20. Slingluff CL, Darrow T, Vevaert C, Quinn-Allen MA, Seigler HF (1988) Human cytotoxic T cells specific for autologous melanoma cells: Successful generation from lymph node cells in seven consecutive cases. J Natl Cancer Inst 80: 1016–1026

    Google Scholar 

  21. Spiess PJ, Yang JC, Rosenberg SA (1987) In vivo antitumor activity of tumor-infiltrating lymphocytes expanded in recombinant interleukin-2. J Natl Cancer Inst 79: 1067–1075

    Google Scholar 

  22. Sporn MB, Roberts AB, Wakefield LM, Assoian RK (1986) Transforming growth factor-β: Biological function and chemical structure. Science 233: 532–534

    Google Scholar 

  23. Takagi S, Chen K, Schwarz RE, Iwatsuki S, Herberman RB, Whiteside TL (1989) Functional and phenotypic analysis of tumorinfiltrating lymphocytes isolated from human primary and metastatic liver tumors and cultured in recombinant interleukin-2. Cancer 63: 102–111

    Google Scholar 

  24. Topalian SL, Solomon D, Rosenberg SA (1989) Tumor-specific cytolysis by lymphocytes infiltrating human melanomas. J Immunol 142: 3714–3725

    Google Scholar 

  25. Trenn G, Takayama H, Hu-Li J, Paul WE, Sitkovsky MV (1988) B cell stimulatory factor I (IL-4) enhances the development of cytotoxic T cells from Lyt-2+ resting muring T lymphocytes. J Immunol 140: 1101–1106

    Google Scholar 

  26. Uchida A, Moore M (1984) Lysis of fresh human tumor cells by autologous large granular lymphocytes and T-lymphocytes: two distinct killing activities induced by coculture with autologous tumor. J Natl Cancer Inst 73: 1285–1292

    Google Scholar 

  27. Vaccarello L, Wang YL, Whiteside TL (1990) Sustained outgrowth of autotumor-reactive T lymphocytes from human solid tumors in the presence of tumor necrosis factor-alpha and interleukin 2. Hum Immunol 28: 216–227

    Google Scholar 

  28. Wang YL, Si L, Kanbour A, Herberman RB, Whiteside TL (1989) Lymphocytes infiltrating human ovarian tumors: synergy between tumor necrosis factor-α and interleukin 2 in the generation of CD8+ effectors from tumor-infiltrating lymphocytes. Cancer Res 49: 5979–5985

    Google Scholar 

  29. Whiteside TL, Miescher S, MacDonald HR, Von Fliedner V (1986) Separation of tumor-infiltrating lymphocytes from tumor cells in human solid tumors. A comparison between velocity sedimentation and discontinuous density gradients. J Immunol Methods 90: 221–233

    Google Scholar 

  30. Whiteside TL, Miescher S, Hurlimann J, Moretta L, Von Fliedner V (1986) Clonal analysis and in situ characterization of lymphocytes infiltrating human breast carcinomas. Cancer Immunol Immunother 23: 169–178

    Google Scholar 

  31. Widmer MB, Grabstein KH (1987) Regulation of cytolytic T-lymphocyte generation by B-cell stimulatory factor. Nature 326: 795–798

    Google Scholar 

  32. Yoo YK, Heo DS, Hata K, Van Thiel DH, Whiteside TL (1990) Tumor-infiltrating lymphocytes from human colon carcinomas. Functional and phenotypic characteristics after long-term culture in recombinant interleukin 2. Gastroenterology 98: 259–268

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Room 7201.2, 203 DeSoto Street, 15 213, Pittsburgh, PA, USA

    Yukihiro Shimizu, Ronald B. Herberman & Theresa L. Whiteside

  2. Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, USA

    Shunzaburo Iwatsuki

  3. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, USA

    Yukihiro Shimizu, Ronald B. Herberman & Theresa L. Whiteside

  4. Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, USA

    Ronald B. Herberman

Authors
  1. Yukihiro Shimizu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shunzaburo Iwatsuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ronald B. Herberman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Theresa L. Whiteside
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by ACS grants IM27 077 and IM588 A (TLW) and Organ Transplant Program Project 1P01-CA-4744501 AZ

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shimizu, Y., Iwatsuki, S., Herberman, R.B. et al. Effects of cytokines on in vitro growth of tumor-infiltrating lymphocytes obtained from human primary and metastatic liver tumors. Cancer Immunol Immunother 32, 280–288 (1991). https://doi.org/10.1007/BF01789045

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01789045

Keywords

Search

Navigation