Skip to main content
PMC home page
Cancer Immunology, Immunotherapy : CII logoLink to Cancer Immunology, Immunotherapy : CII
. 1987 Nov;25(3):161–168. doi: 10.1007/BF00199142

Lysis of fresh murine mammary tumor cells by syngeneic natural killer cells and lymphokine-activated killer cells

Ira H Ames 1,✉, Christopher E Gates 1, A Mariano Garcia 1, Patricia A John 2, Anne K Hennig 2, Russell H Tomar 2
PMCID: PMC11038798  PMID: 3499983

Abstract

We have compared the ability of natural killer (NK) cells from two substrains of C3H mice that differ with respect to their susceptibility to the development of mammary adenocarcinomas to lyse fresh syngeneic mammary tumor cells. Single cell suspensions of mammary tumors from retired breeder females were used as targets in 22-h 51Cr-release cytotoxicity assays with syngeneic NK cells. Tumor cell suspensions were prepared by enzymatic digestion of finely minced tissue followed by centrifugation through a discontinuous Percoll gradient. Effector cells were prepared by passing spleen cells over nylon wool followed by centrifugation through Percoll fraction 7. Syngeneic NK cells had significant levels of lysis against 5/8 tumors studied. NK cells from low risk animals (C3Heb/FeJ) consistently demonstrated greater cytotoxicity against tumor cell preparations than did effectors from the high tumor substrain (C3H/OuJ). Study of cytocentrifuge preparations stained with Wright-Giemsa revealed that the two substrains were identical with respect to the number of azurophilic granules present in the cytoplasm of their NK cells. We have also shown that lymphokine-activated killer (LAK) cells can be generated from splenocytes in C3H mice. While LAK cells from both substrains were capable of lysing fresh syngeneic mammary tumor cells in vitro, LAK cells from the animals at high risk for the formation of mammary adenocarcinomas had greater cytotoxicity against tumor cell suspensions than LAK cells from the low tumor substrain.

Keywords: Natural Killer Cell, Mammary Tumor Cell, Mammary Adenocarcinoma, Tumor Cell Suspension, Murine Mammary Tumor

References

  • 1.Ames IH, Garcia AM, John PA, Litty CA, Farrell MA, Tomar RH. Decreased natural cytotoxicity in mice with high incidence of mammary adenocarcinoma. Clin Immunol Immunopathol. 1986;38:265. doi: 10.1016/0090-1229(86)90144-3. [DOI] [PubMed] [Google Scholar]
  • 2.Barlozzari T, Reynolds CW, Herberman RB. In vivo role of natural killer cells: Involvement of large granular lymphocytes in the clearance of tumor cells in anti-asialo GM1-treated rats. J Immunol. 1983;131:1024. [PubMed] [Google Scholar]
  • 3.Barlozzari T, Leonhardt J, Wiltrout RH, Herberman RB, Reynolds CW. Direct evidence for the role of LGL in the inhibition of experimental tumor metastases. J Immunol. 1985;134:2783. [PubMed] [Google Scholar]
  • 4.Becker S. Intratumor NK reactivity. In: Herberman RB, editor. Natural cell-mediated immunity against tumors. New York: Academic Press; 1980. p. 985. [Google Scholar]
  • 5.Came PE, Moore DH. Inhibition of spontaneous mammary carcinoma of mice by treatment with interferon and poly I:C (35565) Proc Soc Exp Biol Med. 1971;137:304. doi: 10.3181/00379727-137-35565. [DOI] [PubMed] [Google Scholar]
  • 6.Cheever MA, Greenberg PD, Fefer A. Therapy of leukemia by nonimmune syngeneic spleen cells. J Immunol. 1980;124:2137. [PubMed] [Google Scholar]
  • 7.Cikes M, Friberg S, Klein G. Progressive loss of H-2 antigens with concomitant increase of cell-surface antigen(s) determined by Moloney leukemia virus in cultured murine lymphomas. J Natl Cancer Inst. 1973;50:347. doi: 10.1093/jnci/50.2.347. [DOI] [PubMed] [Google Scholar]
  • 8.Dickson C, Peters G. Proteins encoded by mouse mammary tumour virus. Curr Top Microbiol Immunol. 1983;106:1. doi: 10.1007/978-3-642-69357-1_1. [DOI] [PubMed] [Google Scholar]
  • 9.Droller MJ, Gomolka D. Inhibition of tumor growth in association with modification of in vivo immune response by indomethacin and polyinosinic:polycytidylic acid. Cancer Res. 1982;42:5038. [PubMed] [Google Scholar]
  • 10.Droller MJ, Gomolka D. Modulation of stimulatory effects of poly (I:C) on natural cytotoxicity by anti-interferon. Cell Immunol. 1984;86:242. doi: 10.1016/0008-8749(84)90376-9. [DOI] [PubMed] [Google Scholar]
  • 11.Gerson JM. Systemic and in situ natural killer activity in tumor-bearing mice and patients with cancer. In: Herberman RB, editor. Natural cell-mediated immunity against tumors. New York: Academic Press; 1980. p. 1047. [Google Scholar]
  • 12.Gorelik E, Wiltrout RH, Okumura K, Habu S, Herberman RB. Role of NK cells in the control of metastatic spread and growth of tumor cells in mice. Int J Cancer. 1982;30:107. doi: 10.1002/ijc.2910300118. [DOI] [PubMed] [Google Scholar]
  • 13.Gorelik E, Bere WW, Herberman RB. Role of NK cells in the antimetastatic effect of anticoagulant drugs. Int J Cancer. 1984;33:87. doi: 10.1002/ijc.2910330115. [DOI] [PubMed] [Google Scholar]
  • 14.Grimm EA. Human lymphokine-activated killer cells (LAK cells) as a potential immunotherapeutic modality. Biochim Biophys Acta. 1986;865:267. doi: 10.1016/0304-419x(86)90017-x. [DOI] [PubMed] [Google Scholar]
  • 15.Grimm EA, Mazumder A, Zhang HZ, Rosenberg SA. 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;155:1823. doi: 10.1084/jem.155.6.1823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Grimm EA, Ramsey KM, Mazumder A, Wilson DJ, Djeu JY, Rosenberg SA. 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. 1983;157:884. doi: 10.1084/jem.157.3.884. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Habu S, Shimanura K, Akamatsu K-I, Okumura K, Tamaoki N. Protective role of natural killer cells in tumor growth and viral infection in mice. Exp Cell Biol. 1984;52:40. [PubMed] [Google Scholar]
  • 18.Haller O, Kiessling R, Örn A, Wigzell H. Generation of natural killer cells: An autonomous function of the bone marrow. J Exp Med. 1977;145:1411. doi: 10.1084/jem.145.5.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Hanna N. Expression of metastatic potential of tumor cells in young nude mice is correlated with low levels of natural killer cell-mediated cytotoxicity. Int J Cancer. 1980;26:675. doi: 10.1002/ijc.2910260521. [DOI] [PubMed] [Google Scholar]
  • 20.Hanna N, Fidler IJ. Expression of metastatic potential of allogenic and xenogeneic neoplasms in young nude mice. Cancer Res. 1981;41:438. [PubMed] [Google Scholar]
  • 21.Heiniger H-J, Dorey JJ. Handbook on genetically standardized Jax mice. Bar Harbor: The Jackson Laboratory; 1980. [Google Scholar]
  • 22.Herberman RB. Natural killer cells and their possible relevance to transplantation biology. Transplantation. 1982;34:1. [PubMed] [Google Scholar]
  • 23.Herberman RB. Lymphoid cells in immune surveillance against malignant transformation. Adv Host Def Mech. 1983;2:241. [Google Scholar]
  • 24.Herberman RB, Holden HT. Natural cell-mediated immunity. Adv Cancer Res. 1978;27:305. doi: 10.1016/s0065-230x(08)60936-7. [DOI] [PubMed] [Google Scholar]
  • 25.Herberman RB, Ortaldo JR. Natural killer cells: Their role in defenses against disease. Science. 1981;214:24. doi: 10.1126/science.7025208. [DOI] [PubMed] [Google Scholar]
  • 26.Hynes NE, Groner B, Michalides R. Mouse mammary tumor virus: Transcriptional control and involvement in tumorigenesis. Adv Cancer Res. 1984;41:155. doi: 10.1016/s0065-230x(08)60016-0. [DOI] [PubMed] [Google Scholar]
  • 27.Julius MH, Simpson E, Herzenberg LA. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973;3:645. doi: 10.1002/eji.1830031011. [DOI] [PubMed] [Google Scholar]
  • 28.Kärre K, Klein GO, Kiessling R, Klein G, Roder JC. Low natural in vivo resistance to syngeneic leukaemias in natural killer-deficient mice. Nature. 1980;284:624. doi: 10.1038/284624a0. [DOI] [PubMed] [Google Scholar]
  • 29.Kasai M, Leclerc JC, McVay-Boudreau L, Shen FW, Cantor H. Direct evidence that natural killer cells in nonimmune spleen cell populations prevent tumor growth in vivo. J Exp Med. 1979;149:1260. doi: 10.1084/jem.149.5.1260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Kedar E, Chriqui-Zeira E, Mitelman S. Methods for amplifying the induction and expression of cytotoxic response in vitro to syngeneic and autologous freshly-isolated solid tumors of mice. Cancer Immunol Immunother. 1984;18:126. doi: 10.1007/BF00205747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Lacour F, Delage G, Chianale C. Reduced incidence of spontaneous mammary tumors in C3H/He mice after treatment with polyadenylate-polyuridylate. Science. 1975;187:256. doi: 10.1126/science.163037. [DOI] [PubMed] [Google Scholar]
  • 32.Lotze MT, Grimm EA, Mazumder A, Strausser JL, Rosenberg SA. Lysis of fresh and cultured autologous tumor by human lymphocytes cultured in T-cell growth factor. Cancer Res. 1981;41:4420. [PubMed] [Google Scholar]
  • 33.Merluzzi VJ, Savage DM, Souza L, Boone T, Mertelsmann R, Welte K, Last-Barney K. Lysis of spontaneous murine breast tumors by human interleukin 2-stimulated syngeneic T-lymphocytes. Cancer Res. 1985;45:203. [PubMed] [Google Scholar]
  • 34.Nishimura T, Yagi H, Uchiyama Y, Hashimoto Y. Generation of lymphokine-activated killer (LAK) cells from tumor-infiltrating lymphocytes. Cell Immunol. 1986;100:149. doi: 10.1016/0008-8749(86)90015-8. [DOI] [PubMed] [Google Scholar]
  • 35.Nunn ME, Herberman RB, Holden HT. Natural cell-mediated cytotoxicity in mice against non-lymphoid tumor cells and some normal cells. Int J Cancer. 1977;20:381. doi: 10.1002/ijc.2910200309. [DOI] [PubMed] [Google Scholar]
  • 36.Ortaldo JR, Herberman RB. Heterogeneity of natural killer cells. Annu Rev Immunol. 1984;2:359. doi: 10.1146/annurev.iy.02.040184.002043. [DOI] [PubMed] [Google Scholar]
  • 37.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]
  • 38.Oshimi K, Oshimi Y, Yamada O, Mizoguchi H. Lysis of lymphoma cells by autologous and allogeneic natural killer cells. Blood. 1985;65:638. [PubMed] [Google Scholar]
  • 39.Phillips JH, Lanier LL. Dissection of the lymphokine-activated killer (LAK) phenomenon: Relative contribution by NK cells and cytotoxic T lymphocytes. Nat Immu Cell Growth Regul. 1986;5:152. doi: 10.1084/jem.164.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Riccardi C, Barlozzari T, Santoni A, Herberman RB, Cesarini C. Transfer to cyclophosphamide-treated mice of natural killer (NK) cells and in vivo natural reactivity against tumors. J Immunol. 1981;126:1284. [PubMed] [Google Scholar]
  • 41.Risenfeld I, Örn A, Gidlund M, Axberg I, Alm GV, Wigzell H. Positive correlation between in vitro NK activity and in vivo resistance towards AKR lymphoma cells. Int J Cancer. 1980;25:399. doi: 10.1002/ijc.2910250315. [DOI] [PubMed] [Google Scholar]
  • 42.Rosenberg SA. The adoptive immunotherapy of cancer using the transfer of activated lymphoid cells and interleukin-2. Semin Oncol. 1986;13:200. [PubMed] [Google Scholar]
  • 43.Rosenberg EB, Herberman RB, Levine PH, Halterman RH, McCoy IL, Wunderlich JR. Lymphocyte cytotoxicity reactions to leukemia-associated antigens in identical twins. Int J Cancer. 1972;9:648. doi: 10.1002/ijc.2910090323. [DOI] [PubMed] [Google Scholar]
  • 44.Rosenberg SA, Grimm EA, McGrogan M, Doyle M, Kawasaki E, Koths K, Mark DF. Biological activity of recombinant human interleukin-2 produced in Escherichia coli . Science. 1984;223:1412. doi: 10.1126/science.6367046. [DOI] [PubMed] [Google Scholar]
  • 45.Rosenstein M, Yron I, Kaufmann Y, Rosenberg SA. Lymphokine-activated killer cells: Lysis of fresh syngeneic natural killer-resistant murine tumor cells by lymphocytes cultured in interleukin 2. Cancer Res. 1984;44:1946. [PubMed] [Google Scholar]
  • 46.Serrate S, Herberman RB. Natural cell-mediated cytotoxicity against primary mammary tumors. Fed Proc. 1981;40:1007. [Google Scholar]
  • 47.Serrate SA, Vose BM, Timonen T, Ortaldo JR, Herberman RB. Association of human natural killer cell activity against human primary tumors with large granular lymphocytes. In: Herberman RB, editor. NK cells and other natural effector cells. New York: Academic Press; 1982. p. 1055. [Google Scholar]
  • 48.Sheil JM, Gallimore PH, Zimmer SG, Sopori ML. Susceptibility of adenovirus 2-transformed rat cell lines to natural killer (NK) cells: Direct correlation between NK resistance and in vivo tumorigenesis. J Immunol. 1984;132:1578. [PubMed] [Google Scholar]
  • 49.Steel RGD, Torrie JH. Principles and procedures of statistics. New York: McGraw-Hill; 1960. [Google Scholar]
  • 50.Stutman O. Tumor development in mouse strains with low NK activity. Excerpta Med Int Congr Ser. 1984;641:389. [Google Scholar]
  • 51.Strayer DR, Carter WA, Mayberry SD, Pequignot E, Brodsky I. Low natural cytotoxicity of peripheral blood mononuclear cells in individuals with high familial incidences of cancer. Cancer Res. 1984;44:370. [PubMed] [Google Scholar]
  • 52.Talmadge JE, Meyers KM, Prieur DJ, Starkey JR. Role of NK cells in tumour growth and metastasis in beige mice. Nature. 1980;284:622. doi: 10.1038/284622a0. [DOI] [PubMed] [Google Scholar]
  • 53.Timonen T, Reynolds CW, Ortaldo JR, Herberman RB. Isolation of human and rat natural killer cells. J Immunol Meth. 1982;51:269. doi: 10.1016/0022-1759(82)90393-3. [DOI] [PubMed] [Google Scholar]
  • 54.Trinchieri G, Perussia B. Human natural killer cells: Biologic and pathologic aspects. Lab Invest. 1984;50:489. [PubMed] [Google Scholar]
  • 55.Uchida A, Micksche M. Lysis of fresh human tumor cells by autologous large granular lymphocytes from peripheral blood and pleural effusions. Int J Cancer. 1983;32:37. doi: 10.1002/ijc.2910320107. [DOI] [PubMed] [Google Scholar]
  • 56.Uchida A, Yanagawa E. Natural killer cell activity and autologous tumor killing activity in cancer patients: Overlapping involvement of effector cells as determined in two-target conjugate cytotoxicity assay. J Natl Cancer Inst. 1984;73:1093. [PubMed] [Google Scholar]
  • 57.Vánky FT, Argov SA, Einhorn SA, Klein E. Role of alloantigens in natural killing. Allogeneic but not autologous tumor biopsy cells are sensitive for interferon-induced cytotoxicity of human blood lymphocytes. J Exp Med. 1980;151:1151. doi: 10.1084/jem.151.5.1151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 58.Youn JK, Hovanessian AG, Riviere Y, Hue G, Lacour F. Enhancement of natural killer cell activity and 2–5A synthetase in mice treated with polyadenylic-polyuridylic acid. Cell Immunol. 1983;79:298. doi: 10.1016/0008-8749(83)90072-2. [DOI] [PubMed] [Google Scholar]
  • 59.Yron I, Wood TA, Spiess PJ, Rosenberg SA. In vitro growth of murine T cells. V. The isolation and growth of lymphoid cells infiltrating syngeneic solid tumors. J Immunol. 1980;125:238. [PubMed] [Google Scholar]
  • 60.Zagury D, Morgan DA, Fouchard M, Bernard J, Feldman M, Herberman RB. Heterogeneity of human natural killer cells. J Natl Cancer Inst. 1985;74:553. [PubMed] [Google Scholar]
  • 61.Zarling JM, Eskra L, Borden EC, Horoszewicz J, Carter WA. Activation of human natural killer cells cytotoxic for human leukemia cells by purified interferon. J Immunol. 1979;123:63. [PubMed] [Google Scholar]

Articles from Cancer Immunology, Immunotherapy : CII are provided here courtesy of Springer

RESOURCES