Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Jul;57(7):2057–2065. doi: 10.1128/iai.57.7.2057-2065.1989

Selective proliferation of natural killer cells among monocyte-depleted peripheral blood mononuclear cells as a result of stimulation with staphylococcal enterotoxin B.

P Garcia-Peñarrubia 1, M P Lennon 1, F T Koster 1, R O Kelley 1, A D Bankhurst 1
PMCID: PMC313841  PMID: 2731983

Abstract

In vitro stimulation of monocyte-depleted peripheral blood mononuclear cells with staphylococcal enterotoxin B (SEB) resulted in selective proliferation of cells which express the phenotypic and functional characteristics of natural killer (NK) cells. This culture system provides an easy method for obtaining highly purified NK cells, by sequential incubation of monocyte-depleted cells with SEB and then with interleukin-2 (IL-2). After culture for 4 to 5 days in the presence of SEB, 98 to 100% of the cells expressed the CD16 (Leu11) and HNK-1 (Leu19) antigens. This purification occurred through the death of lymphocytes lacking NK cell markers and marked proliferation of NK cells themselves, which leads to an enrichment of the NK cell population. Activation of NK cells was detected by the appearance of the gamma interferon receptor and IL-2 receptor antigens. This homogeneous population showed the morphology of large granular lymphocytes, were potent effectors of cell-mediated cytotoxicity against K562 and Daudi tumor cell lines, and were able to kill gram-positive and gram-negative bacteria. IL-2 was necessary to maintain the activation and proliferation after SEB stimulation for 4 days. Moreover, the maximum frequency of binding to K562 cells (60.6%) was similar to that recently found (58 +/- 3%) (P. Garcia-Peñarrubia, F. T. Koster, and A. D. Bankhurst, J. Immunol. Methods 118:199-208, 1989) with fresh and highly purified NK cells. This method can be used as a source of highly purified NK cells to study their functional properties and applications to the treatment of cancer.

Full text

PDF
2057

Images in this article

2063Image
on p.2063

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abo T., Sugawara S., Amenomori A., Itoh H., Rikiishi H., Moro I., Kumagai K. Selective phagocytosis of gram-positive bacteria and interleukin 1-like factor production by a subpopulation of large granular lymphocytes. J Immunol. 1986 May 1;136(9):3189–3197. [PubMed] [Google Scholar]
  2. Baker P. J., Hiernaux J. R., Fauntleroy M. B., Prescott B., Cantrell J. L., Rudbach J. A. Inactivation of suppressor T-cell activity by nontoxic monophosphoryl lipid A. Infect Immun. 1988 May;56(5):1076–1083. doi: 10.1128/iai.56.5.1076-1083.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buxser S., Bonventre P. F., Archer D. L. Specific receptor binding of staphylococcal enterotoxins by murine splenic lymphocytes. Infect Immun. 1981 Sep;33(3):827–833. doi: 10.1128/iai.33.3.827-833.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Donnelly R. P., Rogers T. J. Immunosuppression induced by staphylococcal enterotoxin B. Cell Immunol. 1982 Sep 1;72(1):166–177. doi: 10.1016/0008-8749(82)90294-5. [DOI] [PubMed] [Google Scholar]
  5. Garcia-Peñarrubia P., Bankhurst A. D., Koster F. T. Experimental and theoretical kinetics study of antibacterial killing mediated by human natural killer cells. J Immunol. 1989 Feb 15;142(4):1310–1317. [PubMed] [Google Scholar]
  6. Garcia-Peñarrubia P., Koster F. T., Bankhurst A. D. The maximum conjugate frequency (alpha max) characterizes killer cell populations. J Immunol Methods. 1989 Mar 31;118(2):199–208. doi: 10.1016/0022-1759(89)90007-0. [DOI] [PubMed] [Google Scholar]
  7. Garcia-Peñarrubia P., Koster F. T., Kelley R. O., McDowell T. D., Bankhurst A. D. Antibacterial activity of human natural killer cells. J Exp Med. 1989 Jan 1;169(1):99–113. doi: 10.1084/jem.169.1.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Glasebrook A. L. Conjugate formation by primary and secondary populations of murine immune T lymphocytes. J Immunol. 1978 Nov;121(5):1870–1877. [PubMed] [Google Scholar]
  9. Greaves M. F., Brown G. Purification of human T and B lymphocytes. J Immunol. 1974 Jan;112(1):420–423. [PubMed] [Google Scholar]
  10. Grimm E. A., Mazumder A., Zhang H. Z., Rosenberg S. A. 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. 1982 Jun 1;155(6):1823–1841. doi: 10.1084/jem.155.6.1823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henkart P. A. Mechanism of lymphocyte-mediated cytotoxicity. Annu Rev Immunol. 1985;3:31–58. doi: 10.1146/annurev.iy.03.040185.000335. [DOI] [PubMed] [Google Scholar]
  12. Herberman R. B., Reynolds C. W., Ortaldo J. R. Mechanism of cytotoxicity by natural killer (NK) cells. Annu Rev Immunol. 1986;4:651–680. doi: 10.1146/annurev.iy.04.040186.003251. [DOI] [PubMed] [Google Scholar]
  13. Kawaguchi-Nagata K., Okamura H., Shoji K., Kanagawa H., Semma M., Shinagawa K. Immunomodulating activities of staphylococcal enterotoxins. I. Effects on in vivo antibody responses and contact sensitivity reaction. Microbiol Immunol. 1985;29(3):183–193. doi: 10.1111/j.1348-0421.1985.tb00818.x. [DOI] [PubMed] [Google Scholar]
  14. Klimpel G. R., Niesel D. W., Asuncion M., Klimpel K. D. Natural killer cell activation and interferon production by peripheral blood lymphocytes after exposure to bacteria. Infect Immun. 1988 Jun;56(6):1436–1441. doi: 10.1128/iai.56.6.1436-1441.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Lindemann R. A. Bacterial activation of human natural killer cells: role of cell surface lipopolysaccharide. Infect Immun. 1988 May;56(5):1301–1308. doi: 10.1128/iai.56.5.1301-1308.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ortaldo J. R., Herberman R. B. Heterogeneity of natural killer cells. Annu Rev Immunol. 1984;2:359–394. doi: 10.1146/annurev.iy.02.040184.002043. [DOI] [PubMed] [Google Scholar]
  18. Ortaldo J. R., Mason A., Overton R. Lymphokine-activated killer cells. Analysis of progenitors and effectors. J Exp Med. 1986 Oct 1;164(4):1193–1205. doi: 10.1084/jem.164.4.1193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Peavy D. L., Adler W. H., Smith R. T. The mitogenic effects of endotoxin and staphylococcal enterotoxin B on mouse spleen cells and human peripheral lymphocytes. J Immunol. 1970 Dec;105(6):1453–1458. [PubMed] [Google Scholar]
  20. Perussia B., Ramoni C., Anegon I., Cuturi M. C., 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(4):171–188. [PubMed] [Google Scholar]
  21. Phillips J. H., Lanier L. L. Dissection of the lymphokine-activated killer phenomenon. Relative contribution of peripheral blood natural killer cells and T lymphocytes to cytolysis. J Exp Med. 1986 Sep 1;164(3):814–825. doi: 10.1084/jem.164.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pinto M., Torten M., Birnbaum S. C. Suppression of the in vivo humoral and cellular immune response by staphylococcal enterotoxin B (SEB). Transplantation. 1978 Jun;25(6):320–323. doi: 10.1097/00007890-197806000-00008. [DOI] [PubMed] [Google Scholar]
  23. Platsoucas C. D., Oleszak E. L., Good R. A. Immunomodulation of human leukocytes by staphylococcal enterotoxin A: augmentation of natural killer cells and induction of suppressor cells. Cell Immunol. 1986 Feb;97(2):371–385. doi: 10.1016/0008-8749(86)90407-7. [DOI] [PubMed] [Google Scholar]
  24. Rosenberg S. A., Lotze M. T., Muul L. M., Leitman S., Chang A. E., Ettinghausen S. E., Matory Y. L., Skibber J. M., Shiloni E., Vetto J. T. Observations on the systemic administration of autologous lymphokine-activated killer cells and recombinant interleukin-2 to patients with metastatic cancer. N Engl J Med. 1985 Dec 5;313(23):1485–1492. doi: 10.1056/NEJM198512053132327. [DOI] [PubMed] [Google Scholar]
  25. Silva A., Bonavida B., Targan S. Mode of action of interferon-mediated modulation of natural killer cytotoxic activity: recruitment of pre-NK cells and enhanced kinetics of lysis. J Immunol. 1980 Aug;125(2):479–484. [PubMed] [Google Scholar]
  26. Smith B. G., Johnson H. M. The effect of staphylococcal enterotoxins on the primary in vitro immune response. J Immunol. 1975 Aug;115(2):575–578. [PubMed] [Google Scholar]
  27. Sondel P. M., Hank J. A., Kohler P. C., Chen B. P., Minkoff D. Z., Molenda J. A. Destruction of autologous human lymphocytes by interleukin 2-activated cytotoxic cells. J Immunol. 1986 Jul 15;137(2):502–511. [PubMed] [Google Scholar]
  28. Tarkkanen J., Saksela E., Lanier L. L. Bacterial activation of human natural killer cells. Characteristics of the activation process and identification of the effector cell. J Immunol. 1986 Oct 15;137(8):2428–2433. [PubMed] [Google Scholar]
  29. Tarkkanen J., Saxén H., Nurminen M., Mäkelä P. H., Saksela E. Bacterial induction of human activated lymphocyte killing and its inhibition by lipopolysaccharide (LPS). J Immunol. 1986 Apr 1;136(7):2662–2669. [PubMed] [Google Scholar]
  30. Tarkkanen J., Timonen T., Saksela E. Induction of autologous reactivity of human NK cells. Scand J Immunol. 1980;11(4):383–390. doi: 10.1111/j.1365-3083.1980.tb00004.x. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Trinchieri G., Perussia B. Human natural killer cells: biologic and pathologic aspects. Lab Invest. 1984 May;50(5):489–513. [PubMed] [Google Scholar]
  33. Yamada S., Ruscetti F. W., Overton W. R., Herberman R. B., Birchenall-Sparks M. C., Ortaldo J. R. Regulation of human large granular lymphocyte and T cell growth and function by recombinant interleukin 2: induction of interleukin 2 receptor and promotion of growth of cells with enhanced cytotoxicity. J Leukoc Biol. 1987 Jun;41(6):505–517. doi: 10.1002/jlb.41.6.505. [DOI] [PubMed] [Google Scholar]
  34. Zehavi-Willner T., Berke G. The mitogenic activity of staphylococcal enterotoxin B (SEB): a monovalent T cell mitogen that stimulates cytolytic T lymphocytes but cannot mediate their lytic interaction. J Immunol. 1986 Oct 15;137(8):2682–2687. [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES