Summary
YAC, a Moloney-virus-induced tumor of A-strain mice, is a nonimmunogenic tumor. Mice injected with the inactivated neoplastic cells and challenged with viable tumor cells did not survive longer than mice that received the challenge dose alone. The homogenate of this nonimmunogenic tumor was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). After electrophoresis, the gel slices containing isolated molecular entities were injected into various groups of mice. The mice were challenged with low doses of viable tumor cells (10–30 cells) and their survival time was recorded. Small but significant numbers of mice injected with apparent 80–90 K SDS-PAGE-isolated molecular entity rejected the tumor or survived longer than the control groups of mice. Spleen cells from mice injected with 80–90 K molecular entity inhibited the YAC tumor cotransferred with them to naive recipients (Winn assay). Spleen cells from mice injected with monoclonal antibody against nonspecific T-cell helper factor and immunized with 80–90 K SDS-PAGE-isolated molecular entity failed to inhibit the tumor growth in naive recipients, indicating that helper T cells are involved in induction of the antitumor resistance. Nylon-wool-passed splenocytes from mice injected with 80–90 K inhibited tumor growth in some of the recipient mice. Spleen cells from these mice treated with anti-Thy-1 and complement also inhibited the tumor growth in some of the recipients, suggesting that the effector cells were both T and non-T cells. C57BL/6 mice immunized with apparent 20 K SDS-PAGE-isolated molecular entity of RBL5 tumor also induced in vivo resistance to the syngeneic viable RBL5 cells, but not to the syngeneic B16 melanoma cells, indicating the specificity of the protective effect. The practical and theoretical implications of these findings are discussed.
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Ahituv A, Naor D, Sharon R, Tarcic N, Klein BY (1982) 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. Cancer Immunol Immunother 14:16
Alaba O, Rogers MJ, Law LW (1979) Rauscher leukemia virus-induced tumor antigens: complete separation from gp70, p30 and H-2. Int J Cancer 24:608
Fidler IJ (1970) Metestasis: Quantitative analysis of distribution and fate of tumor emboli labelled with 125I-5-iodo-2′-deoxyuridine. J Natl Cancer Inst 45:773
Galili N, Devens B, Naor D, Becker S, Klein E (1978) Immune responses to weakly immunogenic virally induced tumors. I. Overcoming low responsiveness by priming mice with a syngeneic in vitro tumor line or allogeneic cross-reactive tumor. Eur J Immunol 8:17
Greene MI, Dorf ME, Pierres M, Benacerraf B (1977) Reduction of syngeneic tumor growth by an anti-I-J alloantiserum. Proc Natl Acad Sci USA 74:5118
James RFL, Kontiainen S, Maudsley DJ, Culbert EJ, Feldmann M (1983) A monoclonal antibody against antigen-specific helper factor augments T-cell help. Nature 301:160
Julius MH, Simpson E, Herzenberg LA (1973) A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol 3:645
Klein BY, Frenkel S, Ahituv A, Naor D (1980) Immunogenicity of subcellular fractions and molecular species of MuLV-induced tumors I. Screening of immunogenic components by isotypic ultracentrifugation and polyacrylamide electrophoresis of a tumor homogenate. J Immunol Methods 38:325
Klein BY, Sharon R, Avraham A, Ahituv A, Naor D (1983) Isolation of immunogenic entities from nonimmunogenic transplantable tumors and their normal cell counterparts. Transplant Proc 15:225
Klein BY, Sharon R, Tarcic N, Naor D (1982) Induction of antitumor reactive cells or suppressor cells by different molecular species isolated from the same nonimmunogenic tumor. Immunobiol 163:7
Klein G, Klein E, Haughton G (1966) Variation of antigenic characteristics between different mouse lymphomas induced by the Moloney virus. J Natl Cancer Inst 36:607
Kobrin BJ, Naor D, Klein BY (1981) 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 the Rauscher-virus-induced RBL5 tumor. J Immunol 126:1874
Lake P, Clark EA, Khorshidi M, Sunshine GH (1979) Production and characterization of cytotoxic Thy-1 antibody-secreting hybrid cell lines. Detection of T cell subsets. Eur J Immunol 9:875
McCoy JL, Fefer A, Glynn JP (1967) comparative studies on the induction of transplantation resistance in BALB/c and C57BL/6 mice in three murine leukemia systems. Cancer Res 27:1743
Naor D (1979) Suppressor cells: permitters and promoters of malignancy? Adv Cancer Res 29:45
Naor D (1983) Coexistence of immunogenic and suppressogenic epitopes in tumor cells and various types of macromolecules. Cancer Immunol Immunother 16:1
Siegel S (1956) Nonparametric statistics for behavioral sciences. The Kolmogorov-Smirnov two-sample test. McGraw-Hill, New York, Kogakusha, Tykyo, International Student Edition, p 127
Tada T, Okumura K (1979) The role of antigen-specific T cell factors in the immune response. Adv Immunol 28:1
Weber K, Osborn M (1975) Proteins and sodium dodecyl sulfate: molecular weight determination on polyacrylamide gels and related procedures. In: Neurath H (ed) The protein, 3rd edn. vol. I. Academic Press, New York, p 179
Winn HJ (1960) The immune response and the homograft reaction. Natl Cancer Inst Monogr 2:113
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Sharon, R., Naor, D. The isolation of immunogenic molecular entities from immunogenic and nonimmunogenic tumor homogenates by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Cancer Immunol Immunother 18, 203–208 (1984). https://doi.org/10.1007/BF00205512
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DOI: https://doi.org/10.1007/BF00205512