Skip to main content
British Journal of Cancer logoLink to British Journal of Cancer
. 1979 Dec;40(6):831–838. doi: 10.1038/bjc.1979.273

Partial purification of tumour-specific transplantation antigens from methylcholanthrene-induced murine sarcomas by immobilized lectins.

K Sikora, G Koch, S Brenner, E Lennox
PMCID: PMC2010135  PMID: 93484

Abstract

Plasma membranes isolated from two immunogenic, non-cross-protecting, MC sarcomas were shown to retain the specific rejection antigens of whole cells as well as serologically detected H-2 antigens. Solubilization of the membranes with sodium deoxycholate gave quantitative release of H-2 and retained the rejection specificity of the tumour from which it was derived. Polyacrylamide-gel electrophoresis (PAGE) showed no extensive degradation of membrane components during solubilization. The solubilized TSTAs were further characterized and purified on columns of 4 different lectins immobilized on sepharose beads. TSTA from both tumours bound to WGA but not to Con A, LCH or RCA columns. Specific activity was retained after elution from the WGA column. Serologically detectable H-2 bound to the Con A and LCH columns only. Clear separation of H-2 from TSTA activity was thus obtained. Furthermore the WGA-binding material represents a source for further purification of TSTA molecules in order to explore the basis for their diversity.

Full text

PDF
831

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Basombrío M. A. Search for common antigenicities among twenty-five sarcomas induced by methylcholanthrene. Cancer Res. 1970 Oct;30(10):2458–2462. [PubMed] [Google Scholar]
  2. Bridgen J., Snary D., Crumpton M. J. Isolation and N-terminal amino acid sequence of membrane-bound human HLA-A and HLA-B antigens. Nature. 1976 May 20;261(5557):200–205. doi: 10.1038/261200a0. [DOI] [PubMed] [Google Scholar]
  3. David G. S., Reisfeld R. A. Binding of carcinoembryonic antigen (CEA) to concanavalin A-sepharose: storage of high-specific-activity 125I-CEA. J Natl Cancer Inst. 1974 Oct;53(4):1005–1010. doi: 10.1093/jnci/53.4.1005. [DOI] [PubMed] [Google Scholar]
  4. DeLeo A. B., Shiku H., Takahashi T., John M., Old L. J. Cell surface antigens of chemically induced sarcomas of the mouse. I. Murine leukemia virus-related antigens and alloantigens on cultured fibroblasts and sarcoma cells: description of a unique antigen on BALB/c Meth A sarcoma. J Exp Med. 1977 Sep 1;146(3):720–734. doi: 10.1084/jem.146.3.720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gurd J. W., Mahler H. R. Fractionation of synaptic plasma membrane glycoproteins by lectin affinity chromatography. Biochemistry. 1974 Dec 3;13(25):5193–5198. doi: 10.1021/bi00722a022. [DOI] [PubMed] [Google Scholar]
  6. Henriksen O., Robinson E. A., Appella E. Structural characterization of H-2 antigens purified from mouse liver. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3322–3326. doi: 10.1073/pnas.75.7.3322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Holmes E. C., Morton D. L., Schidlovsky G., Trahan E. Cross-reacting tumor-specific transplantation antigens in methylcholanthrene-induced guinea pig sarcomas. J Natl Cancer Inst. 1971 Apr;46(4):693–700. [PubMed] [Google Scholar]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Law L. W., Appella E., Henriksen O., Rogers M. Some biologic and biochemical properties of soluble tumor antigens. Ann N Y Acad Sci. 1976;276:11–25. doi: 10.1111/j.1749-6632.1976.tb41633.x. [DOI] [PubMed] [Google Scholar]
  10. Lotan R., Beattie G., Hubbell W., Nicolson G. L. Activities of lectins and their immobilized derivatives in detergent solutions. Implications on the use of lectin affinity chromatography for the purification of membrane glycoproteins. Biochemistry. 1977 May 3;16(9):1787–1794. doi: 10.1021/bi00628a004. [DOI] [PubMed] [Google Scholar]
  11. McCollester D. L. Isolation of meth A cell surface membranes possessing tumor-specific transplantation antigen activity. Cancer Res. 1970 Dec;30(12):2832–2837. [PubMed] [Google Scholar]
  12. Meltzer M. S., Oppenheim J. J., Littman B. H., Leonard E. J., Rapp H. J. Cell-mediated tumor immunity measured in vitro and in vivo with soluble tumor-specific antigens. J Natl Cancer Inst. 1972 Sep;49(3):727–734. [PubMed] [Google Scholar]
  13. Natori T., Law L. W., Appella E. 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. 1977 Sep;37(9):3406–3413. [PubMed] [Google Scholar]
  14. Oettgen H. F., Old L. J., McLean E. P., Carswell E. A. Delayed hypersensitivity and transplantation immunity elicited by soluble antigens of chemically induced tumours in inbred guinea-pigs. Nature. 1968 Oct 19;220(5164):295–297. doi: 10.1038/220295a0. [DOI] [PubMed] [Google Scholar]
  15. Parker G. A., Rosenberg S. A. Serologic identification of multiple tumor-associated antigens on murine sarcomas. J Natl Cancer Inst. 1977 May;58(5):1303–1309. doi: 10.1093/jnci/58.5.1303. [DOI] [PubMed] [Google Scholar]
  16. Pellis N. R., Kahan B. D. Specific tumor immunity induced with soluble materials: restricted range of antigen dose and of challenge tumor load for immunoprotection. J Immunol. 1975 Dec;115(6):1717–1722. [PubMed] [Google Scholar]
  17. Robinson P. J., Bull F. G., Anderton B. H., Roitt I. M. Direct autoradiographic visualisation in SDS-gels of lectin-binding components of the human erythrocyte membrane. FEBS Lett. 1975 Oct 15;58(1):330–333. doi: 10.1016/0014-5793(75)80291-2. [DOI] [PubMed] [Google Scholar]
  18. Sharon N., Lis H. Lectins: cell-agglutinating and sugar-specific proteins. Science. 1972 Sep 15;177(4053):949–959. doi: 10.1126/science.177.4053.949. [DOI] [PubMed] [Google Scholar]
  19. Siegert W., Fenyö E. M., Klein G. Separation of the Moloney leukemia virus-determined cell surface antigen (MCSA) from known virion proteins associated with the cell membrane. Int J Cancer. 1977 Jul 15;20(1):75–82. doi: 10.1002/ijc.2910200113. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES