Summary
Crude 3 M KCl extracts of the methylcholanthrene-induced fibrosarcoma of C3H/HeJ mice, MCA-F, were demonstrated to contain two fractions, one inducing tumor resistance and the other facilitating the outgrowth of neoplastic cell challenge. In immunoprotection tests in syngeneic C3H/HeJ mice, optimal doses of crude solubilized tumor antigen afforded only a 28% reduction in growth compared with saline-treated controls. When crude extracts were fractionated by preparative isoelectric focusing (pIEF) in a slab of superfine Sephadex G-75, significant biologic activity was demonstrated in two fractions. Fraction (Fr) 1, pI 2.5–3.6, induced potent tumor facilitation, increasing the tumor size by more than 100%, while Fr 15, pI 5.8–6.0, engendered resistance that reduced their respective biological effects to MCA-F, but not the antigenically unrelated MCA-D tumor. Thus 3 M KCl extracts contain at least two biologically active components, one immunoprotective and one tumor-facilitating. Since the weak immunoprotective activity of crude materials may represent the vectorial effect of these antagonistic components, subsequent molecular characterization of both moieties may afford insight into the complex response of hosts toward tumors. Furthermore, TSTA purified by the rapid method of isoelectric focusing may be a more suitable reagent for immunotherapy than the parent crude 3 M KCl extracts by virtue of the absence of facilitating antigens.
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Abbreviations
- CE:
-
crude 3 M KCl extract
- pIEF:
-
preparative isoelectric focusing
- Fr:
-
fraction from pIEF
- MCA-F and MCA-D:
-
antigenically different methylcholanthrene-induced fibrosarcomas of C3H/HeJ mice
- TSTA:
-
tumor specific transplantation antigens
References
Baldwin RW (1973) Immunological aspects of chemical carcinogenesis. Adv Cancer Res 18:1
Baldwin RW, Price MF (1975) Neoantigen expression in chemical carcinogenesis. In: Becker FF (ed) Cancer: A comprehensive treatise, vol 1. Plenum Press, New York, p 353
Barra Y, Astier AM, Meyer G (1977) Isolation of polyoma virus-induced antigen. J Natl Cancer Inst 58:721
Blackstock R, Schimpff RD, Smith RT (1978) Heterogeneity of KCl-solubilized antigens of chemically induced sarcomas (Immunology) (40052). Proc Soc Exp Biol Med 157:354
Bowen JG, Baldwin RW (1976) Isolation and characterization of tumour-specific antigen from the serum of rats bearing transplanted aminoazo dye-induced hepatomas. Transplantation 21:213
Brandschaft PB, Aoki T, Silverman TN (1976) Immunogenic and immunosuppressive components in soluble preparations from Maloney murine leukemia virus-induced tumor cells. Int J Cancer 17:678
Burk MW, Yu S, Ristow SS, McKhann CF (1975) Refractoriness of lymphnode cells from tumour-bearing animals. Int J Cancer 15:99
Coggin JH (1979) Induction of transplantation resistance with soluble Simian Virus 40-induced hamster tumor-specific transplantation antigen. Cancer Res 39:2952
Kahan BD, Pellis NR (1975) Specific immunoprotection with 3 M KCl solubilized tumor antigen. J Surg Res 18:263
Law LW, Appella E (1975) Studies of soluble transplantation and tumor antigens. In: Becker FF (ed) Cancer: A comprehensive treatise, vol 4. Plenum Press, New York, p 135
Leonard EJ, Richardson AK, Hardy AS, Rapp HJ (1975) Extraction of tumor-specific antigen from cells and plasma membranes of line-10 hepatoma. J Natl Cancer Inst 55:73
Lowry OH, Rosebrough MJ, Farr AL, Randall RJ (1951) Protein measurement with Folin Phenol Reagent. J Biol Chem 193:265
Luborsky SW, Chang C, Pancake SJ, Mora PT (1976) Detergent solubilized and molecular weight estimation of tumor specific surface antigen from SV40 virus transformed cells. Biochem Biophys Res Commun 71:990
Meltzer MS, Leonard EJ, Hardy AS, Rapp HJ (1975) Protective tumor immunity induced by potassium chloride extracts of guinea pig hepatomas. J Natl Cancer Inst 54:1349
Natori T, Law LW, Appella E (1977) Biological and biochemical properties of nonidet P40-solubilized and partially purified tumor-specific antigens of the transplantation type from plasma membranes of a methylcholanthrene-induced sarcoma. Cancer Res 37:3406
Natori T, Law LW, Appella E (1978) Immunochemical evidence of a tumor-specific surface antigen obtained by detergent solubilization of a chemically induced sarcoma, meth.-A. Cancer Res 38:359
Pellis NR, Kahan BD (1975) Specific immunity induced with soluble materials: restricted range of antigen dose and of challenge tumor load for immunoprotection. J Immunol 115:1717
Pellis NR, Kahan BD (1976) Methods to demonstrate the immunogenicity of soluble tumor specific transplantation antigens: The immunoprophylaxis assay. Methods Cancer Res 13:291
Pellis NR, Kahan BD (1978) Methods to demonstrate the immunogenicity of soluble tumor antigens. II. The local adoptive transfer assay. Methods Cancer Res 14:29
Pellis NR, Tom BH, Kahan BD (1974) Tumor specific and allospecific immunogenicity of soluble extracts from chemically induced murine sarcomas. J Immunol 113:708
Pellis NR, Shulan DJ, Kahan BD (1976) Specific immunity induced with soluble materials: Purification of antigens inducing tumor resistance. Biochem Biophys Res Commun 71:1251
Radola BJ (1974) Isoelectric focusing in layers of granulated gels. II. Preparative isoelectric focusing. Biochim Biophys Acta 386:181
Rao VS, Bagai R, Bonavida B (1976) Specific enhancement of tumor growth and depression of cell-mediated immunity following sensitization to soluble tumor antigens. Cancer Res 36:1384
Reisfeld RA, Kahan BD (1970) Biologic and chemical characterization of human histocompatibility antigens. Fed Proc 29:2034
Rogers MJ, Law LW (1979) Immunogenic properties of a soluble tumor rejection antigen (TSTA) from a Simian Virus 40-induced sarcoma. Int J Cancer 23:89
Šimkovič D, Chorváth B, Duraj J, Hlubinová K (1978) Inhibition and promotion of growth of B77-virus-induced rat tumor with KCl-solubilized tumor cell components. Neoplasma 25:647
Steele G Jr, Sjogren HO, Price MR (1975) Tumor-associated and embryonic antigens in soluble fractions of a chemically-induced rat colon carcinoma. Int J Cancer 16:33
Stevens RH, Brooks GP, Osborne JW, Hoffman KL, Lawson AJ (1978) Lymphocyte cytotoxicity in X-irradiation-induced rat small bowel adenocarcinoma. III. Blocking by 3 M KCl extract. J Immunol 120:335
Vaage J, Jones RD, Brown BW (1972) Tumor-specific resistance in mice detected by inhibition of macrophage migration. Cancer Res 32:680
Yamauchi K, Fujimoto S, Tada T (1979) Differential activation of cytotoxic and suppressor T cells against syngeneic tumors in the mouse. J Immunol 123:1653
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Pellis, N.R., Yamagishi, H., Shulan, D.J. et al. Use of preparative isoelectric focusing in a Sephadex gel slab to separate immunizing and growth facilitating moieties in crude 3 M KCl extracts of a murine fibrosarcoma. Cancer Immunol Immunother 11, 53–58 (1981). https://doi.org/10.1007/BF00205774
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DOI: https://doi.org/10.1007/BF00205774