Skip to main content
SpringerLink
Log in

Phenotypic and functional analysis of lymphokine-activated killer (LAK) cell clones

Ability of CD3+, LAK cell clones to produce interferon-γ and tumor necrosis factor upon stimulation with tumor targets

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

Summary

Lymphokine-activated killer (LAK) cells are generated by the culture of peripheral blood lymphocytes with interleukin-2 (IL-2). A variety of cells, including T-lymphocytes and natural killer (NK) cells, can be activated by IL-2 to exhibit the ability to kill multiple tumor and “modified-self” targets. Recent reports indicate that culture conditions can determine the phenotype of cells expressing LAK activity. Using limiting dilution techniques, we first generated cloned LAK cells with three culture conditions: autologous human serum (AHS)+IL-2; AHS+IL-2+0.1 μg/ml phytohemagglutinin and fetal bovine serum and IL-2. We determined that all but one of the 47 LAK cell clones generated with the three culture conditions were CD3+ and T-cell like; one NK-like clone was observed. Clones that were cytotoxic for one target could generally kill multiple targets, and the absence of phytohemagglutinin did not significantly affect the ability of the LAK cell clones to kill multiple targets. The presence of phytohemagglutinin was, however, necessary for the long-term maintenance of proliferation and cytotoxic activity of the LAK cell clones. The mechanism by which LAK cells kill tumor targets is not known. We here demonstrate that LAK cells and LAK cell clones can produce interferon-γ and tumor necrosis factor (TNF) when stimulated with an erythroleukemia cell, K562. Five of the six CD3+, LAK cell clones tested could be stimulated by K562 cells to produce both interferon-γ and TNF. However, the ability of the cloned LAK cells to kill K562 cells, as measured in a 4-h 51Cr-release assay, did not correlate with their ability to produce these cytokines. Furthermore, specific antibodies that neutralize the cytotoxic activity of interferon-γ and TNF did not inhibit killing of K562 cells by LAK cells as measured with a 4-h cytotoxic assay. The cytostatic and cytotoxic activities of interferon-γ and TNF for tumor cells are well documented, but these cytolytic activities are slower acting and exhibit their maximum effect after 48–96 h. We here propose that LAK cells kill tumor targets by a combination of cell-to-cell-mediated killing and by the release of slower acting cytostatic/cytotoxic cytokines that can inhibit the growth of tumors some distance from the effector cells.

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. Ahrens PB, Ankel H (1987) The role of asparagine-linked carbohydrate in natural killer cell-mediated cytolysis. J Biol Chem 262:7575

    Google Scholar 

  2. Allavena P, Ortaldo JR (1984) Characteristics of human NK clones: target specificity and phenotype. J Immunol 132:2363

    Google Scholar 

  3. Allavena P, Scala G, Djeu JY, Procopio AD, Oppenheim JJ, Herberman RB, Ortaldo JR (1985) Production of multiple cytokines by clones of human large granular lymphocytes. Cancer Immunol Immunother 19:121

    Google Scholar 

  4. Chong AS-F, Hersh EM, Grimes WJ (1988) Blocking of lymphokine activated killer (LAK) cell mediated cytotoxicity by cell sized beads bearing tumor cell proteins. J Immunol 141:4418

    Google Scholar 

  5. Chong AS-F, Scuderi P, Grimes WJ, Hersh EM (1989) Tumor targets stimulate interleukin-2 activated killer (LAK) cells to produce interferon-γ (IFN-γ) and tumor necrosis factor. J Immunol (in press)

  6. Damle NK, Doyle LV, Bradley EC (1986) Interleukin 2-activated human killer cells are derived from phenotypically heterogeneous precursors. J Immunol 137:2814

    Google Scholar 

  7. Ferrini S, Miescher S, Zocchi MR, Von Fliedner V, Moretta A (1987) Phenotypic and functional characterization of recombinant interleukin 2 (rIL-2)-induced activated killer cells: analysis at the population and clonal levels. J Immunol 138:1297

    Google Scholar 

  8. Ferrini S, Moretta L, Pantaleo G, Moretta A (1987) Surface markers of human lymphokine-activated killer cells and their precursors. Analysis at the population and clonal level. Int J Cancer 39:18

    Google Scholar 

  9. Granger GA, Orr SL, Yamamoto RS (1985) Lymphotoxins, macrophage cytotoxins, and tumor necrosis factors: an interrelated family of antitumor effector molecules. J Clin Immunol 5:217

    Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  11. Grimm EA, Robb RJ, Roth JA, Neckers LM, Lachman LB, Wilson DJ, Rosenberg SA (1983) Lymphokine-activated killer cell phenomenon. III. Evidence that IL-2 is sufficient for direct activation of peripheral blood lymphocytes into lymphokine-activated killer cells. J Exp Med 158:1356

    Article  CAS  PubMed  Google Scholar 

  12. Herberman RB, Hiserodt J, Vujanovic N, Balch C, et al. (1987) Lymphokine-activated killer cell activity. Characteristics of effector cells and their progenitors in blood and spleen. Immunol Today 8:178

    Article  Google Scholar 

  13. Hersh EM, Scuderi P, Salmon S, Chong A, Grimes WJ (1988) A soluble anti-tumor factor produced during interleukin-2 (IL-2) induced generation of lymphokine activated killer (LAK) cells. Evidence for a dual mechanism of LAK cell action. Proc Am Assoc Cancer Res 29:98

    Google Scholar 

  14. Itoh K, Shiba K, Shimizu Y, Suzuki R, Kumagai K (1985) Generation of activated killer (AK) cells by recombinant interleukin 2 (rIL-2) in collaboration with interferon-γ (IFN-γ). J Immunol 134:3124

    Google Scholar 

  15. Itoh K, Tilden AB, Kumagai K, Balch CM (1985) Leu-11+ lymphocytes with natural killer (NK) activity are precursors of recombinant interleukin 2 (rIL-2)-induced activated killer (AK) cells. J Immunol 134:802

    Google Scholar 

  16. Lotze MT, Chang AE, Seipp CA, Simpson C, Vetto JT, Rosenberg SA (1986) High dose recombinant interleukin 2 in the treatment of patients with disseminated cancer: Responses, treatment-related morbidity, and histologic findings. J Am Med Assoc 256:3117

    Google Scholar 

  17. Mingari MC, Ferrini S, Pende D, Bottino C, Prigione I, Moretta A, Moretta L (1987) Phenotypic and functional analysis of human CD3+ and CD3− clones with “lymphokine-activated killer” (LAK) activity. Frequent occurrence of CD3+ LAK clones which produce interleukin-2. Int J Cancer 40:495

    Google Scholar 

  18. Moretta L, Pende D, Cozzani R, Merli A, Bagnasco M, Mingari MC (1986) T cell nature of some lymphokine-activated killer (LAK) cells. Frequency analysis of LAK precursors within human T cell populations and clonal analysis of LAK effector cells. Eur J Immunol 16:1623

    Google Scholar 

  19. Mule JJ, Yang J, Shu S, Rosenberg SA (1986) The anti-tumor efficacy of lymphokine-activated killer cells and recombinant interleukin 2 in vivo: direct correlation between reduction of established metastases and cytolytic activity of lymphokine-activated killer cells. J Immunol 136:3899

    Google Scholar 

  20. Nedwin GE, Svedersky LP, Bringman TS, Palladino MA, Goeddel DV (1985) Effect of interleukin 2, interferon-γ, and mitogens on the production of tumor necrosis factors α and β. J Immunol 135:2492

    Google Scholar 

  21. Ortaldo JR, Longo DL (1988) Human natural lymphocyte effector cells: definition, analysis of activity, and clinical effectiveness. J Nat Cancer Inst 80:999

    Google Scholar 

  22. Ortaldo JR, Mason A, Overton R (1986) Lymphokine-activated killer cells. Analysis of progenitors and effectors. J Exp Med 164:1193

    Google Scholar 

  23. Owen-Schaub LB, Gutterman JU, Grimm EA (1988) Synergy of tumor necrosis factor and interleukin 2 in the activation of human cytotoxic lymphocytes: effect of tumor necrosis factor and interleukin 2 in the generation of human lymphokine-activated killer cell cytotoxicity. Cancer Res 48:788

    Google Scholar 

  24. Peters PM, Ortaldo RJ, Shalaby MR, Svondersky LP, Nedwin GE, Bringman TS, Hass PE, Aggarwal BB, Herberman RB, Goeddel DV, Palladino MA (1986) Natural killer-sensitive targets stimulate production of TNF-α but not TNF-β (lymphotoxin) by highly purified human peripheral blood large granular lymphocytes. J Immunol 137:2592

    Google Scholar 

  25. Phillips JH, Lanier LL (1986) Dissection of the lymphokine-activated killer phenomenon. Relative contribution of peripheral blood natural killer cells and T lymphocytes to cytolysis. J Exp Med 164:814

    Google Scholar 

  26. Rayner AA, Grimm EA, Lotze MT, Chu EW, Rosenberg SA (1985) Lymphokine-activated killer (LAK) cells. Analysis of factors relevant to the immunotherapy of human cancer. Cancer 55:1327

    Google Scholar 

  27. Rayner AA, Grimm EA, Lotze MT, Wilson DJ, Rosenberg SA (1985) Lymphokine-activated killer (LAK) cell phenomenon. IV. Lysis by LAK cell clones of fresh human tumor cells from autologous and multiple allogeneic tumors. J Natl Cancer Inst 75:67

    Google Scholar 

  28. Roberts K, Lotze MT, Rosenberg SA (1987) Separation and functional studies of the human lymphokine-activated killer cell. Cancer Res 47:4366

    Google Scholar 

  29. Rosenstein M, Yron I, Kaufmann Y, Rosenberg SA (1984) Lymphokine-activated killer cells: Lysis of fresh syngeneic natural killer-resistant murine tumor cells by lymphocytes cultured in interleukin 2. Cancer Res 44:1946

    Google Scholar 

  30. Ruggiero V, Latham K, Baglioni C (1987) Cytostatic and cytotoxic activity of tumor necrosis factor on human cancer cells. J Immunol 138:2711

    Google Scholar 

  31. Sawada H, Abo T, Sugawara S, Kumagai K (1988) Prerequisite for the induction of lymphokine-activated killer cells from T lymphocytes. J Immunol 140:3668

    Google Scholar 

  32. Scala G, Allavena P, Djeu JY, Kasahara T, Ortadco JR, Herberman RB, Oppenheim JJ (1984) Human large granular lymphocytes are potent producers of interleukin-1. Nature 309:56

    Google Scholar 

  33. Scuderi P, Sterling KE, Raitano AB, Grogan TM, Rippe RA (1987) Recombinant interferon-γ stimulates the production of human tumor necrosis factor in vitro. J Interferon Res 7:164

    Google Scholar 

  34. Shiiba K, Suzuki R, Kawakami K, Ohuchi A, Kumagai K (1986) Interleukin 2-activated killer cells: generation in collaboration with interferon-γ and its suppression in cancer patients. Cancer Immunol Immunother 21:119

    Google Scholar 

  35. Silvennoinen O, Vakkila J, Hurme M (1988) Accessory cells, dendritic cells, or monocytes, are required for the lymphokine-activated killer cell induction from resting T cell but not from natural killer cell precursors. J Immunol 141:1404

    Google Scholar 

  36. Skibber JM, Lotze MT, Muul LM, Uppenkamp IK, Ross W, Rosenberg SA (1987) Human lymphokine-activated killer cells: further isolation and characterization of the precursor and effector cell. Natl Immunol Cell Growth Regul 6:291

    Google Scholar 

  37. Sondel PM, Hank JA, Kohler PC (1987) Status and potential of interleukin-2 for the treatment of neoplastic disease. Oncology 1:41

    Google Scholar 

  38. Sverdersky LP, Nedwin GE, Goeddel DV, Palladino MA (1985) Interferon-γ enhances induction of lymphotoxin in recombinant interleukin 2-stimulated peripheral blood mononuclear cells. J Immunol 134:1604

    Google Scholar 

  39. Taswell C (1981) Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis. J Immunol 126:1614

    Google Scholar 

  40. Tilden AB, Itoh K, Balch CM (1987) Human lymphokine-activated killer (LAK) cells: Identification of two types of effector cells. J Immunol 138:1068

    Google Scholar 

  41. Williamson BD, Carswell EA, Rubin BY, Prendergast JS, Old LJ (1983) Human tumor necrosis factor produced by human B-cell lines: synergistic cytotoxic factor interaction with human interferon. Proc Natl Acad Sci USA 80:5397

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biochemistry, Arizona Cancer Center, University of Arizona, 85724, Tucson, AZ

    Anita S. -F. Chong & William J. Grimes

  2. Department of Microbiology/Immunology, Arizona Cancer Center, University of Arizona, 85724, Tucson, AZ

    Philip Scuderi & Evan M. Hersh

  3. Department of Internal Medicine, Arizona Cancer Center, University of Arizona, 85724, Tucson, AZ, USA

    Anita S. -F. Chong, Alexandre Aleksijevic & Evan M. Hersh

Authors
  1. Anita S. -F. Chong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexandre Aleksijevic
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Philip Scuderi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Evan M. Hersh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. William J. Grimes
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work is supported in part by grants from the Arizona Disease Research Commission (3364-000000-1-1-AP-6621) and the National Institutes of Health (Grants GM 34121, CA-17094 and CA-23074)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chong, A.S.F., Aleksijevic, A., Scuderi, P. et al. Phenotypic and functional analysis of lymphokine-activated killer (LAK) cell clones. Cancer Immunol Immunother 29, 270–278 (1989). https://doi.org/10.1007/BF00199215

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00199215

Keywords

Search

Navigation