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. 1985 Jun 1;161(6):1464–1482. doi: 10.1084/jem.161.6.1464

A model for the differentiation of human natural killer cells. Studies on the in vitro activation of Leu-11+ granular lymphocytes with a natural killer-sensitive tumor cell, K562

PMCID: PMC2187621  PMID: 3159818

Abstract

A subpopulation of low density granular lymphocytes that express the natural killer (NK) cell-associated Leu-11 antigen (IgG Fc receptor) were stimulated directly by coculture with an NK-sensitive tumor cell, K562. T lymphocytes (Leu-11-) responded only weakly when cocultured with K562. The response of Leu-11+ cells apparently did not require exogeneous factors or accessory cells. The K562-activated cells retained expression of Leu-11 antigen, acquired activation antigens, and were highly cytotoxic against NK-sensitive and -insensitive tumor cells. Anti-IL-2 receptor monoclonal antibody minimally inhibited the activation of Leu-11+ cells by K562, but completely inhibited the phytohemagglutinin-induced activation of the Leu-11- cells from the same individual. Leu-11+ cells can be divided into Leu-7-11+ and Leu- 7+11+ subpopulations using anti-Leu-7 antibody. These subsets were separated by two-color fluorescence-activated cell sorting and cocultured with K562. Proliferation by Leu-7-11+ cells was significantly greater than by Leu-11+7+ cells. Leu-7+11- granular lymphocytes and T lymphocytes (Leu-7-11-) typically proliferated only weakly when cocultured with K562. A proportion of the Leu-7-11+ cells acquired Leu-7 antigen after stimulation with K562, whereas the phenotype of Leu-7+11+, Leu-7+11-, and Leu-7-11- subsets was unaffected. These results demonstrate a developmental relationship between the Leu-7-11+ and Leu-7+11+ lymphocytes and suggest that Leu-7 antigen may be expressed late in the differentiation pathway of NK cells. The direct activation of highly purified Leu-11+ cells by coculture with K562 provides an in vitro model with which to study the activation and maturation of human NK cells.

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Selected References

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  1. Abo T., Cooper M. D., Balch C. M. Characterization of HNK-1+ (Leu-7) human lymphocytes. I. Two distinct phenotypes of human NK cells with different cytotoxic capability. J Immunol. 1982 Oct;129(4):1752–1757. [PubMed] [Google Scholar]
  2. Abo T., Miller C. A., Balch C. M. Characterization of human granular lymphocyte subpopulations expressing HNK-1 (Leu-7) and Leu-11 antigens in the blood and lymphoid tissues from fetuses, neonates and adults. Eur J Immunol. 1984 Jul;14(7):616–623. doi: 10.1002/eji.1830140707. [DOI] [PubMed] [Google Scholar]
  3. Abo T., Miller C. A., Gartland G. L., Balch C. M. Differentiation stages of human natural killer cells in lymphoid tissues from fetal to adult life. J Exp Med. 1983 Jan 1;157(1):273–284. doi: 10.1084/jem.157.1.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Allavena P., Ortaldo J. R. Characteristics of human NK clones: target specificity and phenotype. J Immunol. 1984 May;132(5):2363–2369. [PubMed] [Google Scholar]
  5. Domzig W., Stadler B. M., Herberman R. B. Interleukin 2 dependence of human natural killer (NK) cell activity. J Immunol. 1983 Apr;130(4):1970–1973. [PubMed] [Google Scholar]
  6. Gidlund M., Orn A., Wigzell H., Senik A., Gresser I. Enhanced NK cell activity in mice injected with interferon and interferon inducers. Nature. 1978 Jun 29;273(5665):759–761. doi: 10.1038/273759a0. [DOI] [PubMed] [Google Scholar]
  7. Goto M., Zvaifler N. J. Characterization of the killer cell generated in the autologous mixed leukocyte reaction. J Exp Med. 1983 Apr 1;157(4):1309–1323. doi: 10.1084/jem.157.4.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Herberman R. R., Ortaldo J. R., Bonnard G. D. Augmentation by interferon of human natural and antibody-dependent cell-mediated cytotoxicity. Nature. 1979 Jan 18;277(5693):221–223. doi: 10.1038/277221a0. [DOI] [PubMed] [Google Scholar]
  9. Hercend T., Meuer S., Brennan A., Edson M. A., Acuto O., Reinherz E. L., Schlossman S. F., Ritz J. Identification of a clonally restricted 90 kD heterodimer on two human cloned natural killer cell lines. Its role in cytotoxic effector function. J Exp Med. 1983 Nov 1;158(5):1547–1560. doi: 10.1084/jem.158.5.1547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jondal M., Targan S. In vitro induction of cytotoxic effector cells with spontaneous killer cell specificity. J Exp Med. 1978 Jun 1;147(6):1621–1636. doi: 10.1084/jem.147.6.1621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lanier L. L., Benike C. J., Phillips J. H., Engleman E. G. Recombinant interleukin 2 enhanced natural killer cell-mediated cytotoxicity in human lymphocyte subpopulations expressing the Leu 7 and Leu 11 antigens. J Immunol. 1985 Feb;134(2):794–801. [PubMed] [Google Scholar]
  12. Lanier L. L., Le A. M., Phillips J. H., Warner N. L., Babcock G. F. Subpopulations of human natural killer cells defined by expression of the Leu-7 (HNK-1) and Leu-11 (NK-15) antigens. J Immunol. 1983 Oct;131(4):1789–1796. [PubMed] [Google Scholar]
  13. Leonard W. J., Depper J. M., Uchiyama T., Smith K. A., Waldmann T. A., Greene W. C. A monoclonal antibody that appears to recognize the receptor for human T-cell growth factor; partial characterization of the receptor. Nature. 1982 Nov 18;300(5889):267–269. doi: 10.1038/300267a0. [DOI] [PubMed] [Google Scholar]
  14. López-Botet M., Silva A., Rodríguez J., de Landazuri M. O. Generation of T cell blasts with NK-like activity in human MLC: cellular precursors, IL 2 responsiveness, and phenotype expression. J Immunol. 1982 Sep;129(3):1109–1115. [PubMed] [Google Scholar]
  15. Meuer S. C., Acuto O., Hussey R. E., Hodgdon J. C., Fitzgerald K. A., Schlossman S. F., Reinherz E. L. Evidence for the T3-associated 90K heterodimer as the T-cell antigen receptor. Nature. 1983 Jun 30;303(5920):808–810. doi: 10.1038/303808a0. [DOI] [PubMed] [Google Scholar]
  16. Ortaldo J. R., Sharrow S. O., Timonen T., Herberman R. B. Determination of surface antigens on highly purified human NK cells by flow cytometry with monoclonal antibodies. J Immunol. 1981 Dec;127(6):2401–2409. [PubMed] [Google Scholar]
  17. 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 May;130(5):2133–2141. [PubMed] [Google Scholar]
  18. Perussia B., Trinchieri G., Jackson A., Warner N. L., Faust J., Rumpold H., Kraft D., Lanier L. L. The Fc receptor for IgG on human natural killer cells: phenotypic, functional, and comparative studies with monoclonal antibodies. J Immunol. 1984 Jul;133(1):180–189. [PubMed] [Google Scholar]
  19. Phillips J. H., Le A. M., Lanier L. L. Natural killer cells activated in a human mixed lymphocyte response culture identified by expression of Leu-11 and class II histocompatibility antigens. J Exp Med. 1984 Apr 1;159(4):993–1008. doi: 10.1084/jem.159.4.993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Phillips J. H., Warner N. L., Lanier L. L. Correlation of biophysical properties and cell surface antigenic profile of Percoll gradient-separated human natural killer cells. Nat Immun Cell Growth Regul. 1983;3(2):73–86. [PubMed] [Google Scholar]
  21. Poros A., Klein E. Distinction of anti-K562 and anti-allocytotoxicity in in vitro-stimulated populations of human lymphocytes. Cell Immunol. 1979 Aug;46(1):57–68. doi: 10.1016/0008-8749(79)90245-4. [DOI] [PubMed] [Google Scholar]
  22. Rosenberg S. A., Grimm E. A., McGrogan M., Doyle M., Kawasaki E., Koths K., Mark D. F. Biological activity of recombinant human interleukin-2 produced in Escherichia coli. Science. 1984 Mar 30;223(4643):1412–1414. doi: 10.1126/science.6367046. [DOI] [PubMed] [Google Scholar]
  23. Rumpold H., Kraft D., Obexer G., Böck G., Gebhart W. A monoclonal antibody against a surface antigen shared by human large granular lymphocytes and granulocytes. J Immunol. 1982 Oct;129(4):1458–1464. [PubMed] [Google Scholar]
  24. Seeley J. K., Masucci G., Poros A., Klein E., Golub S. H. Studies on cytotoxicity generated in human mixed lymphocyte cultures. II. Anti-K562 effectors are distinct from allospecific CTL and can be generated from NK-depleted T cells. J Immunol. 1979 Sep;123(3):1303–1311. [PubMed] [Google Scholar]
  25. Strassmann G., Bach F. H., Zarling J. M. Depletion of human NK cells with monoclonal antibodies allows the generation of cytotoxic T lymphocytes without NK-like cells in mixed cultures. J Immunol. 1983 Apr;130(4):1556–1560. [PubMed] [Google Scholar]
  26. Svedersky L. P., Shepard H. M., Spencer S. A., Shalaby M. R., Palladino M. A. Augmentation of human natural cell-mediated cytotoxicity by recombinant human interleukin 2. J Immunol. 1984 Aug;133(2):714–718. [PubMed] [Google Scholar]
  27. Svedmyr E., Wigzell H., Jondal M. Sensitization of human lymphocytes against autologous or allogeneic lymphoblastoid cell lines: characteristics of the reactive cells. Scand J Immunol. 1974;3(4):499–508. doi: 10.1111/j.1365-3083.1974.tb01283.x. [DOI] [PubMed] [Google Scholar]
  28. Timonen T., Ortaldo J. R., Herberman R. B. Analysis by a single cell cytotoxicity assay of natural killer (NK) cells frequencies among human large granular lymphocytes and of the effects of interferon on their activity. J Immunol. 1982 Jun;128(6):2514–2521. [PubMed] [Google Scholar]
  29. Timonen T., Ortaldo J. R., Herberman R. B. Characteristics of human large granular lymphocytes and relationship to natural killer and K cells. J Exp Med. 1981 Mar 1;153(3):569–582. doi: 10.1084/jem.153.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Trinchieri G., Matsumoto-Kobayashi M., Clark S. C., Seehra J., London L., Perussia B. Response of resting human peripheral blood natural killer cells to interleukin 2. J Exp Med. 1984 Oct 1;160(4):1147–1169. doi: 10.1084/jem.160.4.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weiss A., Stobo J. D. Requirement for the coexpression of T3 and the T cell antigen receptor on a malignant human T cell line. J Exp Med. 1984 Nov 1;160(5):1284–1299. doi: 10.1084/jem.160.5.1284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. van de Griend R. J., van Krimpen B. A., Ronteltap C. P., Bolhuis R. L. Rapidly expanded activated human killer cell clones have strong antitumor cell activity and have the surface phenotype of either T gamma, T-non-gamma, or null cells. J Immunol. 1984 Jun;132(6):3185–3191. [PubMed] [Google Scholar]

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