Skip to main content
PMC home page
Cancer Immunology, Immunotherapy : CII logoLink to Cancer Immunology, Immunotherapy : CII
. 2013 Jul 9;17(2):117–123. doi: 10.1007/BF00200047

In vitro elicitation of cytotoxic response against a nonimmunogenic murine tumor by allosensitization

Benny Leshem 1,✉, Bracha Gotsman 1, Eli Kedar 1
PMCID: PMC11039133  PMID: 6235909

Abstract

The murine lymphoma (thymoma) PIR-2 of C57BL/6 origin, primarily induced in our laboratory by fractionated X-ray irradiation, has been shown to be nonimmunogenic by its failure to immunize syngeneic mice in vivo or to evoke a cytotoxic response in primary mixed lymphocyte-tumor cell cultures (MLTC) in vitro. We were able, however, to demonstrate the existence of anti-PIR-2 cytotoxic cells among allogeneic-primed C57BL/6 responding lymphocytes using the technique of limiting dilution cultures (LDC). The frequency of anti-PIR-2 cytotoxic cells among C57BL/6 lymphocytes sensitized against BALB/c splenocytes in mixed leukocyte culture (MLC) was 1/20 to 1/40, and the cytotoxic activity of positive LDC wells against PIR-2 reached 60% as determined by a 4-h 51Cr-release assay. The frequency of anti-PIR-2 cytotoxic cells could be increased two- to 10-fold (up to 1/4) by removing nylon-wool-adherent cells from the primed cell population and/or by enriching the primed lymphoblast population on a Percoll density gradient. Anti-PIR-2 cytotoxic cells were found to be Thy1+; Lytl−2+ cells. Clones isolated from the LDC wells manifested strong cytotoxic activity toward PIR-2 cells and the stimulating BALB/c splenocytes but not against other H-2b tumor lines or C57BL/6 splenocytes. We suggest that the procedure of allostimulation in MLC-LDC is an effective in vitro means of generating highly reactive cytotoxic cells against poorly immunogenic neoplasms.

Keywords: Thymoma, Cytotoxic Cell, Cytotoxic Response, Percoll Density Gradient, Limit Dilution Analysis

References

  • 1.Aoki T, Herberman RB, Hartley JW, Liu M, Walling NJ, Nunn M. Surface antigens on transplantable tumor cell-lines producing mouse type C viruses. J Natl Cancer Inst. 1977;58:1065. doi: 10.1093/jnci/58.4.1069. [DOI] [PubMed] [Google Scholar]
  • 2.Bach ML, Bach FH, Zarling JM. Pool priming: a means of generating T lymphocytes cytotoxic to tumor or virus infected cells. Lancet. 1978;I:20. doi: 10.1016/S0140-6736(78)90364-1. [DOI] [PubMed] [Google Scholar]
  • 3.Bach FH, Leshem B, Kuperman OJ, Bunzendahl H, Alter BJ, Zarling JM. Allosensitization to obtain anti-tumor immunity. In: Fefer A, Goldstein A, editors. The potential role of T cells in cancer therapy. New York: Raven; 1982. p. 7. [Google Scholar]
  • 4.Cerottini JC, Brunner KT. Cell-mediated cytotoxicity, allograft rejection, and tumor immunity. Adv Immunol. 1974;18:67. doi: 10.1016/S0065-2776(08)60308-9. [DOI] [PubMed] [Google Scholar]
  • 5.Finke JH, Fyfe DA, Del Villano BC, Butler GH, Ponzio NM. Characterization of “foreign” alloantigen-like specificities on a murine lymphoma cell line. Transplant Proc. 1980;12:53. [PubMed] [Google Scholar]
  • 6.Finn OJ, Lieberman M, Kaplan HS. H-2 Antigen expression: loss in vitro, restoration in vivo and correlation with cell-mediated cytotoxicity in a mouse lymphoma cell line. Immunogenetics. 1978;7:79. doi: 10.1007/BF01843991. [DOI] [PubMed] [Google Scholar]
  • 7.Fossati G, Balsari A, Taramelli D, Sensi ML, Pellegris G, Nava M, Parmiani G. Lysis of autologous human melanoma cells by in vitro allosensitized peripheral blood lymphocytes. Cancer Immunol Immunother. 1982;14:99. doi: 10.1007/BF00200176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Gately MK, Glaser M, Dick SJ, Mettetal RW, Jr, Kornblith PL. In vitro studies on the cell-mediated immune response to human brain tumors. I. Requirement for third-party stimulator lymphocytes in the induction of cell-mediated cytotoxic responses to allogeneic cultured gliomas. J Natl Cancer Inst. 1982;69:1245. [PubMed] [Google Scholar]
  • 9.Gillis S, Ferm MM, Ou W, Smith KA. T cell growth factor: parameters of production and a quantitative microassay for activity. J Immunol. 1978;120:2027. [PubMed] [Google Scholar]
  • 10.Haas M. Continuous production of radiation leukemia virus in C57BL thymoma tissue culture lines: purification of the leukemogenic virus. Cell. 1974;1:79. doi: 10.1016/0092-8674(74)90065-8. [DOI] [Google Scholar]
  • 11.Hurrell SM, Zarling JM. Ly2+ effectors cytotoxic for syngeneic tumor cells: generation by allogeneic stimulation and by supernatants from mixed leukocyte cultures. J Immunol. 1983;131:1017. [PubMed] [Google Scholar]
  • 12.Ihle JN, Joseph DR, Pazmino NH. Radiation leukemia in C57BL/6 mice. II. Lack of ecotropic virus expression in the majority of lymphomas. J Exp Med. 1976;144:1406. doi: 10.1084/jem.144.6.1406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.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]
  • 14.Kedar E, Weiss DW. The in vitro generation of effector lymphocytes and their employment in tumor immunotherapy. Adv Cancer Res. 1983;38:171. doi: 10.1016/S0065-230X(08)60190-6. [DOI] [PubMed] [Google Scholar]
  • 15.Lee SK, Oliver RTD. Autologous leukemia-specific T-cell mediated lymphocytotoxicity in patients with acute myelogenous leukemia. J Exp Med. 1978;147:912. doi: 10.1084/jem.147.3.912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Leshem B, Kedar E (1983) In vitro elicitation of cytotoxic response against a nonimmunogenic murine tumor. Proceedings of the First Meeting of the Israel Society for the Life Sciences, Jerusalem. Abstract A 198
  • 17.Lieber MM, Sherr CJ, Todaro GS. S-tropic murine type-C viruses: frequency of isolation from continuous cell lines, leukemia virus preparations and normal spleens. Int J Cancer. 1974;13:587. doi: 10.1002/ijc.2910130503. [DOI] [PubMed] [Google Scholar]
  • 18.Lieberman M, Kaplan HS. Leukemogenic activity of filtrates from radiation induced lymphoid tumors of mice. Science. 1959;130:387. doi: 10.1126/science.130.3372.387. [DOI] [PubMed] [Google Scholar]
  • 19.Lieberman M, Kaplan HS, DeCléve A. Anomalous viral expression in radiogenic lymphomas of C57BL/Ka mice. In: Yuhas JM, Tennant RW, Regan JD, editors. Biology of radiation carcinogenesis. New York: Raven; 1976. p. 237. [Google Scholar]
  • 20.MacDonald HR, Cerottini JC, Ryser JE, Maryanski JL, Taswell C, Widmer MB, Brunner KT. Quantitation and cloning of cytolytic T lymphocytes and their precursors. Immunol Rev. 1980;51:93. doi: 10.1111/j.1600-065X.1980.tb00318.x. [DOI] [PubMed] [Google Scholar]
  • 21.Mazumder A, Grimm EA, Rosenberg SA. Lysis of fresh human solid tumors by autologous lymphocytes activated in vitro by allosensitization. Cancer Immunol Immunother. 1983;15:1. doi: 10.1007/BF00199454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Paciucci PA, MacPhail S, Zarling JM, Bach FH. Lysis of syngeneic solid tumor by alloantigen stimulated mouse T and non-T cells. J Immunol. 1980;124:370. [PubMed] [Google Scholar]
  • 23.Sensi ML, Parenza M, Parmiani G. Alloreactivity and tumor antigens: generation of syngeneic anti-lymphoma killer lymphocytes by alloimmunization of mice with normal cells. J Natl Cancer Inst. 1983;70:291. [PubMed] [Google Scholar]
  • 24.Strausser JL, Mazumder A, Grimm EA, Lotze MT, Rosenberg SA. Lysis of human solid tumors by autologous cells sensitized in vitro to alloantigens. J Immunol. 1981;127:266. [PubMed] [Google Scholar]
  • 25.Truitt RL, Shih CY, Lefever AV, Tempelis LD, Andreani M, Bortin MM. Characterization of alloimmunization-induced T lymphocytes reactive against AKR leukemia in vitro and correlation with graft-vs-leukemia activity in vivo. J Immunol. 1983;131:2050. [PubMed] [Google Scholar]
  • 26.Vánky F, Gorsky T, Gorsky Y, Masucci MG, Klein E. Lysis of tumor biopsy cells by autologous T lymphocytes activated in mixed cultures and propagated with T cell growth factor. J Exp Med. 1982;155:83. doi: 10.1084/jem.155.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Vose BM, White W. Tumor reactive lymphocytes stimulated in mixed lymphocyte and tumor culture: clonal analysis of effector cells in cytotoxic and proliferative assays. Cancer Immunol Immunother. 1983;15:227. doi: 10.1007/BF00199170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Yefenof E, Klein G. Antibody induced redistribution of normal and tumor associated surface antigens. Exp Cell Res. 1974;88:217. doi: 10.1016/0014-4827(74)90234-1. [DOI] [PubMed] [Google Scholar]
  • 29.Yefenof E, Tchakirov R, Kedar E. In vitro generation of cytotoxic lymphocytes against radiation and radiation leukemia virus-induced tumors. I. Role of viral antigenicity. Cancer Immunol Immunother. 1980;8:171. [Google Scholar]
  • 30.Zarling JM, Robins HI, Raich PC, Bach FH, Bach ML. Generation of cytotoxic T lymphocytes to autologous human leukemia cells by sensitization to pooled allogeneic normal cells. Nature. 1978;274:269. doi: 10.1038/274269a0. [DOI] [PubMed] [Google Scholar]
  • 31.Zarling JM, Kay NE, Grant B, Yasukawa M, Bach FH. Human anti lymphoma responses generated in vitro and in vivo following sensitization with allogeneic leukocytes. Cancer Immunol Immunother. 1983;15:237. doi: 10.1007/BF00199171. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES