Abstract
The effect of feeder cells on oncolytic activity of lymphocyte subsets and their growth was evaluated in long-term human bone marrow interleukin-2 (IL-2) cultures. Two B-lymphoblastoid cell lines (Daudi and Epstein-Barr-virus-transformed BSM) and two human leukemias, AML-M5, were used as feeder cells. The most prominent effects were seen in cultures stimulated with Daudi cells. In these cultures, cytotoxic activity was 100–1000 times increased against a broad range of target cells and the total cellular expansion was more than 40 times higher than in control cultures. This Daudi-related effect appeared to be mediated by natural killer (NK) cells, since cellular expansion occurred mostly in the CD16+ and CD56+ CD3− NK cell subset. In cultures stimulated with BSM and acute myelogenous leukemia (AML) feeder cells, the increase in proliferation was similar, but the enhancement of cytotoxicity, even though significant, was less prominent. Although all feeder cells were effective in stimulation of bone marrow reactivity, the highest cytotoxicity was always observed with feeder cells autologous to the targets, indicating some degree of specificity. This was especially evident in cultures stimulated with autologous versus allogeneic AML feeder cells. In contrast to Daudistimulated IL-2 cultures, in which the highest expansion of CD3− CD56+ NK cells was observed, in BSM and AML cultures, the CD3+ CD56+/- T cell subsets were more prolific. This indicates that the response and phenotypic heterogeneity of bone marrow cultures depends on the type of feeder cells used. This observation indicates that the preferential stimulation of a pertinent lymphocyte subset for therapeutic purposes may be possible.
Key words: Feeder cells, Bone marrow cultures, IL-2, Lymphocyte subsets
Footnotes
Recipient of Florence Maude Thomas Cancer Research Professorship
References
- 1.Agah R, Malloy B, Kerner M, Mazumder A. Generation and characterization of IL-2 activated bone marrow cells as a potent graft vs tumor effector in transplantation. J Immunol. 1989;143:3093. [PubMed] [Google Scholar]
- 2.Dixon WJ (1982) Biomedical Computer Programs (BMDP), Statistical Software, University of California Press
- 3.Fuchshuber RP, Lotzová E, Pollock RE. Identification of human peripheral blood lymphocytes (PBL) responsible for longterm IL-2 dependent antitumor cytotoxicity. Proc 80th Am Assoc Cancer Res. 1989;30:339. [Google Scholar]
- 4.Kobayashi N, Fritz L, Ryan M, Hewick RM, Clark SC, Chan S, Loedon R, Sherman F, Perussia B, Trinchieri G. Identification and purification of natural killer cell stimulatory factor (NKSF), a cytokine with multiple biologic effects on human lymphocytes. J Exp Med. 1989;170:827. doi: 10.1084/jem.170.3.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Lotzová E. Centrifugal elutriation allows enrichment of natural killing and separates xenogeneic and allogeneic reactivity. In: Herberman RB, editor. Natural cell-mediated immunity against tumors. New York: Academic Press; 1980. pp. 131–137. [Google Scholar]
- 6.Lotzová E. Cytotoxicity and clinical application of activated NK cells. Med Oncol Tumor Pharmacother. 1989;6:93. doi: 10.1007/BF02985229. [DOI] [PubMed] [Google Scholar]
- 7.Lotzová E, Ades EW. Natural killer cells: definition, heterogeneity, lytic mechanism, function and clinical application. Nat Immun Cell Growth Regul. 1989;8:1. [PubMed] [Google Scholar]
- 8.Lotzová E, Herberman RB. Reassessment of LAK phenomenology: a review. Nat Immun Cell Growth Regul. 1987;6:109. [PubMed] [Google Scholar]
- 9.Lotzová E, Savary CA. Generation of NK cell activity from human bone marrow. J Immunol. 1987;139:279. [PubMed] [Google Scholar]
- 10.Lotzová E, Savary CA, Herberman RB. Induction of NK cell activity against fresh leukemia in culture with interleukin-2. J Immunol. 1987;138:2718. [PubMed] [Google Scholar]
- 11.Ortaldo JR, Mason A, Overton R. Lymphokine-activated killer cells. Analysis of progenitors and effectors. J Exp Med. 1986;164:1193. doi: 10.1084/jem.164.4.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12.Perussia B, Starr S, Abraham S, Fanning V, Trinchieri G. Human natural killer cells analyzed by B73.1, a monoclonal antibody blocking Fc receptor functions: I. Characterization of the lymphocyte subset reactive with B73.1. J Immunol. 1983;130:2133. [PubMed] [Google Scholar]
- 13.Perussia B, Ramoni C, Anegon I, Cuturi MC, Faust J, Trinchieri G. Preferential proliferation of natural killer cells among peripheral blood mononuclear cells cocultured with B lymphoblastoid cell lines. Nat Immun Cell growth Regul. 1987;6:171. [PubMed] [Google Scholar]
- 14.Phillips JH, Lanier LL. Dissection of the lymphokine-activated killer phenomenon: relative contribution of peripheral blood natural killer cells and T lymphocytes to cytolysis. J Exp Med. 1986;164:814. doi: 10.1084/jem.164.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Pollock RE, Zimmerman SO, Fuchshuber P, Stanford SD, Lotzová E. Current methods for deriving lytic unit values from human cytotoxicity data. Is there a better way. J Clin Lab Analysis. 1990;4:274–282. doi: 10.1002/jcla.1860040408. [DOI] [PubMed] [Google Scholar]
- 16.Timonen T, Ortaldo JR, Herberman RB. Characteristics of human large granular lymphocytes and relationship to natural killer and K cells. J Exp Med. 1981;155:569. doi: 10.1084/jem.153.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17.Trinchieri G, Perussia B. Human natural killer cells: biologic and pathologic aspects. Lab Invest. 1984;50:489. [PubMed] [Google Scholar]
- 18.Vanky F, Klein E. Auto-tumor lysis by blood lymphocytes in vitro. Strongly activated lymphocytes lack selectivity. Cancer Immunol Immunother. 1989;29:125. doi: 10.1007/BF00199287. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19.Yamada S, Ortaldo JR. Regulation of LGL's and human T cell growth and function by rIL-2. J Leukocyte Biol. 1987;42:263. doi: 10.1002/jlb.42.3.263. [DOI] [PubMed] [Google Scholar]