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. 1982 Nov;14(1):16–26. doi: 10.1007/BF00199427

Immunogenicity of subcellular fractions and molecular species of MuLV-induced tumors

III. Stimulation of syngeneic antitumor responses by subcellular fractions and molecular species of moloney virus-induced tumors in CBA and a mice

A Ahituv 1, D Naor 1, R Sharon 1, N Tarcic 1, B Y Klein 1
PMCID: PMC11039193  PMID: 6186365

Abstract

YBA, a Moloney virus-induced leukemia in CBA mice, and a relatively weak immunogenic tumor, was screened for the presence of immunogenic antigens. The tumor was subjected to homogenization and subcellular fractionation on sucrose gradients; the immunogenic subcellular fractions underwent further separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunogenicity of the subcellular fractions and the SDS-PAGE-isolated molecular species were tested by (their) subcutaneous injection into syngeneic mice and examination of their splenocytes examined against tumor cell and normal cell targets by the chromium release cell-mediated lympholysis assay. Tumor cell homogenates were also separated by SDS-PAGE and tested for immunogenicity without prior fractionation.

Splenocytes from mice that had received injections of certain SDS-PAGE-isolated epitopes derived from YBA tumor homogenate or its light and heavy subcellular fractions generated effective cytotoxic responses against YBA target cells after 6 days in vitro cultivation. In contrast, intact YBA tumor cells or non-separated tumor homogenates failed to induce an efficient cytotoxic response. The effector cells induced with the immunogenic SDS-PAGE-isolated epitopes of YBA tumor were specific, since they cytolysed the homologous target cells more efficiently than unrelated target cells or syngeneic normal cells. The activity of these effector cells was affected by varying the effector : target ratio. Augmentation of the cytotoxic responses was obtained when the splenocytes of mice immunized with SDS-PAGE-isolated epitopes of YBA tumor were restimulated in vitro, with the homologous neoplastic cells.

Immunogenic SDS-PAGE epitopes were isolated from YAC tumor also (YAC is a Moloney-induced tumor of A mice). The effector cells induced with these separated epitopes were characterized as thymus-derived cells and not as natural killer cells.

The results suggest that (1) the molecular repertoire of YBA and YBA tumors contain immunogens that can induce a specific antitumor cell-mediated response; (2) the isolated molecular species injected are more efficient immunogens than the entire, unseparated homogenate sample or a dose of 108 intact inactivated tumor cells; and (3) the gel matrix may be responsible for the enhanced cell-mediated response induced against the weakly immunogenic tumor.

Keywords: Molecular Species, Subcellular Fraction, Cytotoxic Response, Immunogenic Tumor, Tumor Homogenate

References

  • 1.Beverley PCL, Woody J, Dunkley M, Feldmann M. Separation of suppressor and killer T cells by surface phenotype. Nature. 1976;262:495. doi: 10.1038/262495a0. [DOI] [PubMed] [Google Scholar]
  • 2.Boon T, Van Snick J, Van Pel A, Uyttenhove C, Merchand M. Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. II. Lymphocyte-mediated cytolysis. J Exp Med. 1980;152:1184. doi: 10.1084/jem.152.5.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Brunner KT, Mauel J, Cerottini J-C, Chapuis B. Quantitative assay of the lytic action of immune lymphoid cells on 51Cr-labelled allogeneic target cells in vitro; inhibition by isoantibody and by drugs. Immunology. 1968;14:181. [PMC free article] [PubMed] [Google Scholar]
  • 4.Devens B, Galili N, Deutsch O, Naor D, Klein E. Immune responses to weakly immunogenic virally induced tumors. II. Suppressive effects of the in vivo carried tumor YAC. Eur J Immunol. 1978;8:578. doi: 10.1002/eji.1830080807. [DOI] [PubMed] [Google Scholar]
  • 5.Devens B, Schochot L, Naor D. Immune responses to weakly immunogenic virally induced tumors. V. Short in vitro cultivation of YAC changes its antigenic properties. Cell Immunol. 1979;44:442. doi: 10.1016/0008-8749(79)90019-4. [DOI] [PubMed] [Google Scholar]
  • 6.Galili N, Devens B, Naor D, Becker S, Klein E. Immune responses to weakly immunogenic virally induced tumors. I. Overcoming low responsiveness by priming mice with a syngeneic in vitro tumor line or allogeneic crossreactive tumor. Eur J Immunol. 1978;8:17. doi: 10.1002/eji.1830080105. [DOI] [PubMed] [Google Scholar]
  • 7.Glynn JP, McCoy JL, Fefer A. Cross-resistance to the transplantation of syngeneic Friend, Moloney and Rauscher virus-induced tumors. Cancer Res. 1968;28:434. [PubMed] [Google Scholar]
  • 8.Goldstein P, Luciani M-F, Wagner H, Rollinghoff M. Mouse T cell-mediated cytolysis specifically triggered by cytophilic xenogeneic serum determinants: a caveat for the interpretation of experiments done under ‘syngeneic’ conditions. J Immunol. 1978;121:2533. [PubMed] [Google Scholar]
  • 9.Hewitt HB, Blake ER, Walder AS. A critique of the evidence for active host defense against cancer, based on personal studies of 27 murine tumors of spontaneous origin. Br J Cancer. 1976;33:241. doi: 10.1038/bjc.1976.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.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]
  • 11.Kasai M, Iwamori M, Nagai Y, Okumura K, Tada T. A glycolipid on the surface of mouse natural killer cells. Eur J Immunol. 1980;10:175. doi: 10.1002/eji.1830100304. [DOI] [PubMed] [Google Scholar]
  • 12.Kasai M, Yoneda T, Habu S, Maruyama Y, Okumura K, Tokunaga T. In vivo effect of anti-asialo GM1 antibody on natural killer activity. Nature. 1981;291:334. doi: 10.1038/291334a0. [DOI] [PubMed] [Google Scholar]
  • 13.Klein BY, Frenkel S, Ahituv A, Naor D. Immunogenicity of subcellular fractions and molecular species of MuLV-induced tumors. I. Screening of immunogenic components by isopycnic ultracentrifugation and polyacrylamide electrophoresis of a tumor homogenate. J Immunol Methods. 1980;38:325. doi: 10.1016/0022-1759(80)90281-1. [DOI] [PubMed] [Google Scholar]
  • 14.Klein BY, Devens B, Deutsch O, Ahituv A, Frenkel S, Kobrin BJ, Naor D. Isolation of immunogenic and suppressogenic determinants of the nonimmunogenic YAC tumor and the change in its immunogenic repertoire after in vitro cultivation. Transplant Proc. 1981;13:790. [PubMed] [Google Scholar]
  • 15.Klein G, Klein E, Haughton G. Variation of antigenic characteristics between different mouse lymphomas induced by the Moloney virus. J Natl Cancer Inst. 1966;36:607. doi: 10.1093/jnci/36.4.607. [DOI] [PubMed] [Google Scholar]
  • 16.Kobrin BJ, Naor D, Klein BY. Immunogenicity of subcellular fractions and molecular species of MuLV-induced tumors. II. Stimulation of syngeneic anti-tumor cell mediated immune responses by subcellular fractions and molecular species of Rauscher-virus-induced RBL5 tumor. J Immunol. 1981;126:1874. [PubMed] [Google Scholar]
  • 17.Lake P, Clark EA, Khorshidi M, Sunshine GH. Production and characterization of cytotoxic Thy-1 antibody-secreting hybrid cell lines. Detection of T cell subsets. Eur J Immunol. 1979;9:875. doi: 10.1002/eji.1830091109. [DOI] [PubMed] [Google Scholar]
  • 18.Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem. 1951;193:265. [PubMed] [Google Scholar]
  • 19.McCoy JL, Fefer A, Glynn JP. Comparative studies on the induction of transplantation resistance in BALB/C and C57BL/6 mice in three murine leukemia systems. Cancer Res. 1976;27:1743. [PubMed] [Google Scholar]
  • 20.Sharon R, Klein BY, Naor D (1982) T cell and cancer. Proceedings of the XIth Triennial World Congress of Pathology (in press)
  • 21.Weber K, Osborn M. Proteins and sodium dodecyl sulfate: molecular weight determination on polyacrylamide gel and related procedures. In: Neurath H, editor. The proteins. New York: Academic Press; 1975. p. 179. [Google Scholar]
  • 22.Yefenof E, Azar Y, Eidelsztein P, Ben-Sasson SZ. Immortalization of antigen-specific helper T cell lines by transformation with the Radiation Leukemia virus (RadLV) J Immunol. 1982;128:625. [PubMed] [Google Scholar]

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