Cancer Immunology, Immunotherapy

, Volume 32, Issue 4, pp 245–250 | Cite as

Activation and expansion of tumour-infiltrating lymphocytes by anti-CD3 and anti-CD28 monoclonal antibodies

  • Erik W. P. Nijhuis
  • Elly v/d Wiel-van Kemenade
  • Carl G. Figdor
  • René A. W. van Lier
Original articles


Cytotoxic T lymphocytes from healthy donors can be expanded to high numbers from the peripheral blood using combinations of anti-CD3 and anti-CD28 monoclonal antibodies (mAb). We investigated whether these antibodies could also be used to induce outgrowth of tumour-infiltrating lymphocytes (TIL) from tumour tissue. In the initiation phase of TIL culture immobilized anti-CD3 antibodies together with anti-CD28 mAb and low-dose interleukin-2 induced a rapid expansion of T cells from various human tumour tissues. The cultured cells showed high levels of cytotoxic T lymphocyte activity, but low levels of lymphokine-activated killer cell activity were generated. This study shows that TIL can be efficiently expanded from tumour tissue by combinations of anti-CD3 and anti-CD28 antibodies. This protocol for cell expansion in vitro may substantially reduce the time required to reach sufficient numbers of TIL for re-infusion to the patient.


Cancer Research Tumour Tissue Cell Activity Human Tumour Killer Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    Ames IH, Gagne GM, Garcia AM, John PA, Scatorchia GD, Tomar RH, and McAfee JG (1989) Preferential homing of tumor-infiltrating lymphocytes in tumor bearing mice. Cancer Immunol Immunother 29: 93–100Google Scholar
  2. 2.
    Belldegrun A, Muul LM, and Rosenberg SA (1988) Interleukin 2 expanded tumor-infiltrating lymphocytes in human renal cell cancer: isolation, characterization, and antitumor activity. Cancer Res 48: 206–214Google Scholar
  3. 3.
    Bom-Noorloos AA, van Beek AAM, Melief CJM (1980) Cryopreservation of cells for immunological typing of Non-Hodgkin Lymphoma. Cancer Res 40: 2890–2894Google Scholar
  4. 4.
    de Jong R, Brouwer M, Rebel VI, van Seventer GA, Miedema F, and van Lier RAW (1990) Generation of alloreactive cytolytic T lymphocytes by immobilized anti-CD3 monoclonal antibodies. Immunology (in press)Google Scholar
  5. 5.
    Figdor CG, Bont WE, Touw I, de Roos J, Roosnek EE, de Vries JE (1982) Isolation of functionally different human monocytes by counterflow centrifugation elutriation. Blood 60: 46–53Google Scholar
  6. 6.
    Geppert TD, Lipsky PF (1987) Accessory cell independent proliferation of human T4 cells stimulated by immobilized antibodies to CD3. J Immunol 138: 1660–1666Google Scholar
  7. 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: 1419–1441Google Scholar
  8. 8.
    Knuth A, Woelfel T, Klehman E, Boon T, Meyer zum Buschenfelde KH (1989) Cytolytic T-cell clones against an autologous human melanoma: specificity study and definition of three antigens by immunoselection. Proc Natl Acad Sci USA 86: 2804–2808Google Scholar
  9. 9.
    Ledbetter JA, Martin PJ, Spooner E, Wofsy D, Tsu TT, Beatty PG, Gladstone PG (1985) Antibodies to Tp67 and Tp44 augment and sustain proliferative responses of activated T cells. J Immunol 135: 2331–2336Google Scholar
  10. 10.
    Lesslauer W, Koning F, Ottenhof T, Giphart M, Goulmy E, van Rood JJ (1986) T90/44 (9.3 antigen). A cell surface molecule with a function in human T cell activation. Eur J Immunol 16: 1289–1296Google Scholar
  11. 11.
    Lindsten T, June CH, Ledbetter JA, Stella G, Thompson GB (1989) Regulation of lymphokine messenger RNA stability by a surface-mediated T-cell activation pathway. Science 244: 339–343Google Scholar
  12. 12.
    Miesher S, Stoeck M, Qiao L, Barras C, Barrelet L, Von Fliedner V (1988) Proliferative and cytolytic potentials of purified human tumor-infiltrating T lymphocytes. Impaired response to mitogendriven stimulation despite T-cell receptor expression. Int J Cancer 42: 659–666Google Scholar
  13. 13.
    Mukherji B, Guha A, Chakraborty NG, Sivanandham M, Nashed AL, Sporn JR, Ergin MT (1989) Clonal analysis of cytotoxic and regulatory T cell responses against human melanoma. J Exp Med 169: 1961–1976Google Scholar
  14. 14.
    Muul LM, Spiess P, Director EP, Rosenberg SA (1987) Identification of specific cytolytic immune responses against autologous tumor in humans bearing malignant melanoma. J Immunol 138: 989–995Google Scholar
  15. 15.
    Natali PG, Nicotra MR, Bigotti A, Venturo I, Marcenaro L, Giacomini P, Russo C (1989) Selective changes in expression of HLA class I polymorhic determinants in human solid tumors. Proc Natl Acad Sci USA 86: 6719–6723Google Scholar
  16. 16.
    Pisani RJ, Leibson PJ, McKean DJ (1989) In vitro activation of lymphocytes from nonsmall cell cancer patients by interleukin 2 and anti-CD3 antibody. Clin Immunol Immunopathol 50: 348–363Google Scholar
  17. 17.
    Radrizzani M, Gambacorti-Passerini C, Parmiani G, Fossati G (1989) Lysis by interleukin 2-stimulated tumor-infiltrating lymphocytes of autologous and allogeneic tumor cells. Cancer Immunol Immunother 28: 67–73Google Scholar
  18. 18.
    Ranges GE, Figari IS, Espevik T, Palladino MA (1987) Inhibition of cytotoxic T cell development by transforming growth factor beta and reversal by tumor necrosis factor alpha. J Exp Med 166: 991–1006Google Scholar
  19. 19.
    Rosenberg SA, Spiess P, Lafreniere R (1986) A new approach to adoptive immunotherapy of cancer with tumor infiltrating lymphocytes. Science 233: 1318–1321Google Scholar
  20. 20.
    Schoof DD, Jung SE, Eberlein TJ (1989) Human tumor-infiltrating lymphocyte (TIL) cytotoxicity facilitated by anti-T-cell receptor antibody. Int J Cancer 44: 219–224Google Scholar
  21. 21.
    Topalian SL, Solomaon D, Avis FP, Chang AE, Freerksen DL, Lineham WM, Lotze MT, Robertson CN, Seipp CA, Simon P, Simpson CG, Rosenberg SA (1988) Immunotherapy of patients with advanced cancer using tumor-infiltrating lymphocytes and recombinant interleukin-2: a pilot study. J Clin Oncol 6: 839–853Google Scholar
  22. 22.
    van Lier RAW, Brouwer M, Aarden LA (1988) Signals involved in T-cell activation: T-cell proliferation induced through the synergistic action of anti-CD2 and anti-CD28 monoclonal antibodies. Eur J Immunol 18: 167–172Google Scholar
  23. 23.
    van Lier RAW, Brouwer M, de Jong R, Groot M, de Groot E, Aarden LA (1989) Functional properties of the human T-cell differentiation antigen CD28. In: Knapp W, Dorken B, Gilks WR, Rieber EP, Schmidt RE, Stein H, von dem Borne AEGKr (eds) Leucocyte typing IV. Oxford University Press, Oxford, p 353Google Scholar
  24. 24.
    van Lier RAW, Brouwer M, Rebel VI, van Noesel CJM, Aarden LA (1989) Immobilized anti-CD3 monoclonal antibodies induce accessory cell independent lymphokine production, proliferation and helper activity in human T lymphocytes. Immunology 68: 45–50Google Scholar
  25. 25.
    Weiss A, Manger B, Imboden J (1986) Synergy between the T3/antigen receptor complex and Tp44 in the activation of human T cells. J Immunol 137: 819–825Google Scholar
  26. 26.
    Woelfel T, Klehman E, Muller C, Schutt K-H, zum Buschenfelde K-H Meyer, Knuth A (1989) Lysis of human melanoma cells by autologous cytolytic T cell clones. J Exp Med 170: 797–810Google Scholar
  27. 27.
    Yun Y SD, Hargrove ME, Ting CC (1989) In vivo antitumor activity of anti-CD3-induced activated killer cells. Cancer Res 49: 4770–4774Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Erik W. P. Nijhuis
    • 1
  • Elly v/d Wiel-van Kemenade
    • 2
  • Carl G. Figdor
    • 2
  • René A. W. van Lier
    • 1
  1. 1.c/o Publication SecretariatCentral Laboratory of the Netherlands Red Cross Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology of the University of AmsterdamAmsterdamThe Netherlands
  2. 2.Dutch Cancer InstituteAmsterdamThe Netherlands

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