Skip to main content
SpringerLink
Log in

Tumour-reactive lymphocytes stimulated in mixed lymphocyte and tumour culture

Clonal analysis of effector cells in cytotoxic and proliferative assays

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

Summary

Lymphocytes from cancer patients were stimulated in mixed culture with autologous tumour (MLTC) or pooled allogeneic lymphocytes (MLC). Both protocols induced increased uptake of 3H-thymidine at 5 days and the appearance of lymphoblasts. Blasts were isolated on discontinuous Percoll gradients and either expanded as bulk cultures or cloned directly under limiting dilution conditions in the presence of conditioned medium containing IL-2. Results with MLTC-blast-CTC have been reported elsewhere. MLC-activated cultures lysed autologous tumour but not autologous lymphoblasts. Lysis of some allogeneic tumours, lymphoblasts from members of the inducing pool, and K562 was also apparent. MLC activated cultures did not undergo restimulation in response to autologous tumour or lymphocytes but were restimulated by leukocytes from pool members.

MLTC clones showed autologous tumour-specific cytotoxic activity or cross-reactive proliferative responses with tumours of the same site and histology. The majority of MLC clones cytotoxic for autologous tumour were also specific and did not lyse allogeneic tumour, K562, or lymphoblasts from the inducing pool. Two clones lysed autologous tumour and pool members. None of the clones tested proliferated in response to autologous tumour following MLC activation but some were responsive to pool members and one clone was restimulated by autologous monocytes. No association was found between clone phenotype and function. The implication of these data is that the effector cells with activity against autologous tumour induced in MLC arose largely by transstimulation of in vivo-activated tumour reactive lymphocytes by IL-2 release rather than expansion of NK-like effectors or sharing of antigenic specificities between tumour and allogeneic lymphocytes. Since MLC activation of cancer patients lymphocytes does not induce proliferative responses to autologous tumour it is unlikely to be a useful procedure in preparing cells for immunotherapy protocols.

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.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bach FH, Bach ML, Zarling JM (1978) Pool-priming: A means of generating T lymphocytes cytotoxic to tumour or virus-infected cells. Lancet I:20

    Google Scholar 

  2. Bach FH, Leshem B, Kuperman OJ, Bunzendahl H, Alter BJ, Zarling JM (1982) Allosensitisation to obtain anti-tumour immunity. In: Fefer A, Goldstein A (eds) The potential role of T cells in cancer therapy. Raven Press, New York

    Google Scholar 

  3. Dallman MJ, Mason DW (1982) Role of thymus-derived and thymus-independent cells in murine skin allograft rejection. Transplantation 33:221

    Google Scholar 

  4. Fernandez-Cruz E, Woda BA, Feldman JD (1980) Elimination of syngeneic sarcomas in rats by a subset of T lymphocytes. J Exp Med 152:823

    Google Scholar 

  5. Fossati G, Baksari A, Taramelli D, Sensi ML, Pellegris G, Nava M, Parmiani G (1983) Lysis of autologous human melanoma cells by in vitro allosensitised peripheral blood lymphocytes. Cancer Immunol Immunother (in press)

  6. Greenberg PD, Cheever MA, Fefer A (1981) Eradication of disseminated murine leukaemia by chemoimmunotherapy with cyclophosphamide and adaptively transferred immune syngeneic Lyt 1+2− lymphocytes. J Exp Med 154:952

    Google Scholar 

  7. Grimm EA, Mazumder A, Zhang HZ, Rosenberg SA (1982) Lymphokine-activated killer cell phenomenon. Lysis of natural killer resistant fresh solid tumour cells by interleukin-2-activated autologous human peripheral blood lymphocytes. J Exp Med 155:1823–1841

    Article  CAS  PubMed  Google Scholar 

  8. Julius MH, Simpson E, Herzenberg LA (1973) Rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol 3:645–649

    Google Scholar 

  9. Lotze MT, Grimm EA, Mazumder A, Strausser JL, Rosenberg SA (1981) In vitro growth of cytotoxic human lymphocytes. IV. Lysis of fresh and cultured autologous tumour by lymphocytes cultured in T cell growth factor. Cancer Res 41:4420

    Google Scholar 

  10. Loveland BE, MacKenzie IFC (1982) Which T cells cause graft rejection? Transplantation 33:217

    Google Scholar 

  11. Mazumder A, Grimm EA, Zhang HZ, Rosenberg SA (1982) Lysis of fresh human solid tumours by autologous lymphocytes activated in vitro with lectins. Cancer Res 42:913

    Google Scholar 

  12. Ortaldo JR, Timonen TT, Vose BM, Alvarez JA (1982) Human natural killer cells as well as T cells maintained in continuous cultures with IL-2. In: Fefer A, Goldstein A (eds) The potential role of T cells in cancer therapy. Raven Press, New York, p 191

    Google Scholar 

  13. Pawelec GP, Hadam MR, Ziegler, A, Lohmeyer J, Rehbein A, Kumbier I, Wernet P (1982) Long-term culture, cloning and surface markers of mixed leukocyte culture-derived human T lymphocytes with natural killer-like cytotoxicity. J Immunol 128:1892

    Google Scholar 

  14. Strausser JL, Mazumder A, Grimm EA, Lotze MT, Rosenberg SA (1981) Lysis of human solid tumours by autologous cells sensitised in vitro to alloantigens. J Immunol 127:226

    Google Scholar 

  15. Vánky F, Klein E (1982) Specificity of auto-tumour cytotoxicity exerted by fresh, activated and propagated human T lymphocytes. Int J Cancer 29:547

    Google Scholar 

  16. Vánky F, Stjernswärd J (1979) Lymphocyte stimulation (by autologous tumour biopsy cells). In: Herberman RB, McIntire KR (eds) Immunodiagnosis of cancer, part 2. Marcel Dekker, New York, p 998

    Google Scholar 

  17. Vánky F, Klein E, Stjernswärd J, Nilsonne U, Rodriguez L, Peterffy A (1979a) Human tumour-lymphocyte interaction in vitro. II. Conditions which improve the capacity of biopsy cells to stimulate autologous lymphocytes. Cancer Immunol Immunother 5:63

    Google Scholar 

  18. Vánky F, Vose BM, Fopp M, Klein E (1979b) Human tumour-lymphocyte interactions in vitro. VI. Specificity of primary and secondary autologous lymphocyte-mediated cytotoxicity. J Natl Cancer Inst 62:1407

    Google Scholar 

  19. Vánky F, Gorsky T, Gorsky Y, Masucci M-G, Klein E (1982) Lysis of tumour biopsy cells by autologous T lymphocytes activated in mixed cultures and propagated with T cell growth factor. J Exp Med 155:83–95

    Google Scholar 

  20. Vose BM (1978) Cytotoxicity of adherent cells associated with some human tumours and lung tissues. Cancer Immunol Immunother 5:173

    Google Scholar 

  21. Vose BM (1980) Specific T-cell-mediated killing of autologous lung tumour cells. Cell Immunol 55:12

    Google Scholar 

  22. Vose BM (1982a) Quantitation of proliferative and cytotoxic precursor cells directed against human tumours: Limiting dilution analysis in peripheral blood and at the tumour site. Int J Cancer 30:135

    Google Scholar 

  23. Vose BM (1982b) Separation of tumour and host cell populations from human neoplasms. In: Reid E (ed). Methodological surveys, vol II. Ellis Harwood, Chichester p 45

    Google Scholar 

  24. Vose BM, Bonnard GD (1982a) Specific cytotoxicity against autologous tumour and proliferative responses of human lymphocytes grown in interleukin 2. Int J Cancer 29:33

    Google Scholar 

  25. Vose BM, Bonnard GD (1982b) Limiting dilution analysis of the precursor cells capable of specific cytotoxic or proliferative responses to human tumour cells and their expansion with lectin-free interleukin-2. In: Serron B (ed) Current concepts in human immunology and cancer immunomodulation. Elsevier Biomedical Press, Amsterdam, p 385

    Google Scholar 

  26. Vose BM, Bonnard GD (1983) Limiting dilution analysis of the frequency of human T cells and large granular lymphocytes proliferating in response to interleukin-2. I. The effect of lectin on the proliferative and cytotoxic activity of cultured lymphoid cells. J Immunol (in press)

  27. Vose BM, Howell A (1983) Cultured human antitumour T cells and their potential for therapy. In: Herberman RB (ed) Clinical and basic tumour immunology. Martinus Hyhoff, The Hague (in press)

    Google Scholar 

  28. Vose BM, Moore M (1979) Suppressor cell activity of lymphocytes infiltrating human lung and breast tumours. Int J Cancer 24:579

    Google Scholar 

  29. Vose BM, Moore M (1981) Cultured human T-cell lines kill autologous tumour. Immunol Lett 3:237

    Google Scholar 

  30. Vose BM, Vánky F, Klein E (1977) Lymphocyte cytotoxicity against autologous tumour biopsy cells in humans. Int J Cancer 20:512

    Google Scholar 

  31. Vose BM, Vánky F, Fopp M, Klein E (1978a) In vitro generation of cytotoxicity against autologous human tumour biopsy cells. Int J Cancer 21:588

    Google Scholar 

  32. Vose BM, Vánky F, Fopp M, Klein E (1978b) Restricted autologous lymphocytotoxicity in lung neoplasia. Br J Cancer 38:375

    Google Scholar 

  33. Vose BM, Gallagher P, Moore M, Schofield PF (1981) Specific and non-specific lymphocyte cytotoxicity in colon cancer. Br J Cancer 44:846

    Google Scholar 

  34. Vose BM, Riccardi C, Marchmont RJ, Bonnard GD (1983) Clonal analysis of human natural killer cells. In: Herberman RB (ed) NK cells and other natural effector cells. Academic Press, New York, p 879

    Google Scholar 

  35. Zarling JM, Robins HI, Raich PC, Bach FH, Bach ML (1978) Generation of cytotoxic T lymphocytes to autologous human leukaemia cells by sensitisation to pooled allogeneic normal cells. Nature 274:269

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Immunology, Paterson Laboratories, Christie Hospital and Holt Radium Institute, M20 9BX, Manchester, England

    B. M. Vose & W. White

Authors
  1. B. M. Vose
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. W. White
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Abbreviations used in this paper: PBL, peripheral blood lymphocytes; TIL, tumour infiltrating lymphocytes; MLTC, mixed lymphocyte tumour culture; IL-2, interleukin-2; MLC, mixed lymphocyte culture; LSM, lymphocyte separation medium; BSS, balanced salt solution; HuSe, human serum; PBS, phosphate-buffered saline; CTC, cultured T cells; PHA, phytohaemagglutinin; CM, cultured medium; NK, natural killer; FcR, receptor for the Fc portion of IgG

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vose, B.M., White, W. Tumour-reactive lymphocytes stimulated in mixed lymphocyte and tumour culture. Cancer Immunol Immunother 15, 227–236 (1983). https://doi.org/10.1007/BF00199170

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00199170

Keywords

Search

Navigation