Abstract
A series of 62 lymphokine-activated killer cell (LAK) cultures from 44 different donors was investigated for the distribution of various CD markers during a cultivation period of 3 weeks. Great differences in the phenotypic pattern were found between different donors, but similar changes of the subset pattern of various donors allowed a classification of the LAK cultures into four distinct LAK types. LAK type 1 was characterised by low numbers of CD3+ cells and high values for CD56+ cells. In LAK type 2 cultures γ/δTCR+ cells extensively proliferated, whereas in LAK type 3 cultures the CD57 and CD8 values increased considerably. LAK type 4 cultures did not show any of these characteristics. The resulting phenotype of a LAK culture was donor-specific, as LAK cultures established from the same peripheral blood mononuclear cells (PBMC), fresh or after cryopreservation, or from PBMC obtained from the same donor at different venous punctures, always developed the same phenotype. A clear correlation between phenotype and killing activity could only be found for LAK type 1 cultures, which always developed high lytic activity. Long-term IL-2 stimulation induced high levels of perforin-positive cells in LAK cultures but the perforin content did not correlate with the cytotoxicity. The transcription pattern for various cytokines only varied slightly between the cultures. Messenger RNA for granulocyte/macrophage-colony-stimulating factor, interferon γ, tumour necrosis factor α, interleukin-4 (IL-4) and IL-5 were found in almost all cultures during the entire cultivation period, whereas mRNA for IL-2 was never detected. Most variations in the transcription pattern were observed for IL-6 and IL-7. However, no correlation could be found between the endogenous cytokine production and the phenotype or lytic activity of the LAK cultures. Further studies are required to determine the factors that cause lymphocyte subsets from a specific donor to proliferate preferentially under long-term IL-2 stimulation.
Similar content being viewed by others
References
Autran B, Leblond V, Sadat-Sowti B, Lefranc E, Got P, Sutton L, Binet JL, Debré P (1991) A soluble factor released by CD8+CD57+ lymphocytes from bone marrow transplanted patients inhibits cell-mediated cytolysis. Blood 77: 2237
Bergmann L, Weidmann E, Bungert B, Hechler P, Mitrou PS (1990) Influence of various cytokines on the induction of lymphokine-activated killer cells. Nat Immum Cell Growth Regul 9: 265
Crump WL, Owen-Schaub LB, Grimm EA (1989) Human recombinant interleukin-1 is synergistic with interleukin-2 in the generation of lymphokine-activated killer cells. Cancer Res 49: 149
Damle NK, Doyle LV, Bradley EC (1986) Interleukin 2-activated human killer cells are derived from phenotypically hetergeneous precursors. J Immunol 137: 2814
Ebina N, Gallardo D, Shau H, Golub SH (1990) IL-1 and IL-4 as reciprocal regulators of IL-2 induced lymphocyte cytotoxicity. Br J Cancer 62: 619
Farace F, Pallardy M, Angevin E, Hercend T, Escudier B, Triebel F (1994) Metastatic renal-cell carcinoma patients treated with interleukin 2 or interleukin 2 plus interferon γ: immunological monitoring. Int J Cancer 57: 814
Findley HW, Nasr S, Afify Z, Hnath R, Waldrep K, Ragab AH (1990) Effects of recombinant interferon-gamma and interleukin-2 on the generation of lymphokine-activated killer cells in vitro. Cancer Invest 8: 493
Fisch P, Malkovsky M, Braakman E, Sturm E, Bolhuis RLH, Prieve A, Sosman JA, Lam VA, Sondel PM (1990) γ/δ T cell clones and natural killer cell clones mediate distinct patterns of non-major histocompatibility complex-restricted cytolysis. J Exp Med 171: 1567
Fuchshuber PR, Lotzova E, Pollock RE (1991) Antitumor activity, growth, and phenotype of long-term IL-2 cultures of human NK and T lymphocytes. Lymphokine Cytokine Res 10: 51
Gallagher G, Stimson WH, Findlay J, Al-Azzawi F (1990) Interleukin-6 enhances the induction of human lymphokine-activated killer cells. Cancer Immunol Immunother 31: 49
Grimm EA, Mazumder A, Zhang HZ, Rosenberg SA (1982) Lymphokine activated killer cell phenomenon. Lysis of natural killer cell resistant fresh solid tumor cells by interleukin-2 activated autologous human peripheral blood lymphocytes. J Exp Med 155: 1823
Grimm EA, Ramsey KM, Mazumder A, Wilson DJ, Djeu JY, Rosenberg SA (1983) Lymphokine activated killer cell phenomenon. II. Precursor phenotype is serologically distinct from peripheral T lymphocytes, memory cytotoxic thymus-derived lymphocytes, and natural killer cells. J Exp Med 157: 884
Hameed A, Gohlsen KJ, Cheng L, Fox WM, Hruban RH, Podack ER (1992) Immunhistochemical identification of cytotoxic lymphocytes using human perforin monoclonal antibody. Am J Pathol 140: 1025
Herberman RB, Hiserodt J, Vujanovic N, Balch C, Lotzova E, Bolhuis R, Golub S, Lanier LL, Phillips JH, Riccardi C, Ritz J, Santoni A, Schmidt RE, Uchida A (1987) Lymphokine-activated killer cell activity. Characteristics of effector cells and their progenitors in blood and spleen. Immunol Today 8: 178
Kaur I, Voss SD, Gupta RS, Schell K, Fisch P, Sondel PM (1993) Human peripheral γδ T cells recognize hsp60 molecules on Daudi Burkitt's lymphoma cells. J Immunol 150: 2046
Kjeldsen-Kragh J, Quayle AJ, Skalhegg BS, Sioud M, Forre O (1993) Selective activation of resting human γδ T lymphocytes by interleukin-2. Eur J Immunol 23: 2092
Koberda J, Bergmann L, Mitrou PS, Hoelzer D (1991) High release of tumor necrosis factor α, interferon γ and interleukin-6 by adherent lymphokine-activated killer cells phenotypically derived from T cells. J Cancer Res Clin Oncol 117: 425
Kovacs EJ, Beckner SK, Longo DL, Varesio L, Young HA (1989) Cytokine gene expression during the generation of human lymphokine-activated killer cells: early induction of interleukin 1β by interleukin 2. Cancer Res 49: 940
Krähenbühl O, Tschopp J (1991) Perforin-induced pore formation. Immunol Today 12: 399
Limb GA, Meager A, Woolley J, Wadhwa M, Biggerstaff J, Brown KA (1989) Release of cytokines during generation of lymphokine-activated killer (LAK) cells by IL-2. Immunology 68: 514
Mehta S, Flanagan P, Blackinton D, Wanebo H (1992) Lymphokine-activated effector cells: modulation of activity by cytokines. Lymphokine Cytokine Res 11: 73
Moser R, Groscurth P, Fehr J (1990) Promotion of transendothelial neutrophil passage by human thrombin. J Cell Sci 96: 737
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 α and interleukin 2 in the generation of human lymphokine-activated killer cell cytotoxicity. Cancer Res 48: 788
Quan CP, Watanabe S, Vuillier F, Pires R, Matsuo T, Stanislawski M, Pillot J, Bouvet JP (1993) Purification and partial amino acid sequence of suppressive lymphokine from a CD8+CD57+ human T hybridoma. Immunology 78: 205
Roussel E, Gerrard JM, Greenberg AH (1990) Long-term cultures of human peripheral blood lymphocytes with recombinant human interleukin-2 generate a population of virtually pure CD3+CD16−CD56− large granular lymphocyte LAK cells. Clin Exp Immunol 82: 416
Sadat-Sowti B, Debré P, Idziorek T, Guillon JM, Hadida F, Okzenhendler E, Katlama C, Mayaud C, Autran B (1991) A lectin-binding soluble factor released by CD8+CD57+ lymphocytes from AIDS patients inhibits T cell cytotoxicity. Eur J Immunol 21: 737
Saraya KA, Balkwill FR (1993) Temporal sequence and cellular origin of interleukin-2 stimulated cytokine gene expression. Br J Cancer 67: 514
Singh SM, Sone S, Inamura N, Ogura T (1989) Up-regulation by granulocyte-macrophage colony-stimulating factor (GM-CSF) of induction of lymphokine (IL-2)-activated killer (LAK) cells by human blood monocytes. Int J Cancer 44: 170
Spagnoli GC, Juretic A, Schultz-Thater E, Dellabona P, Filgueira L, Hörig H, Zuber M, Garotta G, Heberer M (1993) On the relative role of interleukin-2 and interleukin-10 in the generation of lymphokine-activated killer cell activity. Cell Immunol 146: 391
Sturm E, Braakman E, Fisch P, Vreugdenhil RJ, Sondel P, Bolhius RLH (1990) Human Vγ2 T cell receptor-γδ lymphocytes show specificity to Daudi Burkitt's lymphoma cells. J Immunol 145: 3202
Testa U, Care A, Montesoro E, Fossati C, Giannella G, Masciulli R, Fagioli M, Bulgarini D, Habetswallner D, Isacchi G, Pelicci PG, Peschle C (1990) Interleukin-2-dependent long-term cultures of low-density lymphocytes allow the proliferation of lymphokineactivated killer cells with natural killer, Tiγ/δ or TNK phenotype. Cancer Immunol Immunother 31: 11
Vollenweider I, Vrbka E, Fierz W, Groscurth P (1993) Heterogeneous binding and killing behaviour of human γ/δTCR+ lymphokine-activated killer cells against K562 and Daudi cells. Cancer Immunol Immunother 36: 331
Author information
Authors and Affiliations
Additional information
This work was supported by the Sasselia Foundation, Zürich
Rights and permissions
About this article
Cite this article
Vollenweider, I., Moser, R. & Groscurth, P. Development of four donor-specific phenotypes in human long-term lymphokine-activated killer cell cultures. Cancer Immunol Immunother 39, 305–312 (1994). https://doi.org/10.1007/BF01519983
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01519983