Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1979 Feb 1;149(2):473–484. doi: 10.1084/jem.149.2.473

Selective affinity fractionation of murine cytotoxic T lymphocytes (CTL). Unique lectin specific binding of the CTL associated surface glycoprotein, T 145

PMCID: PMC2184808  PMID: 310865

Abstract

We have analyzed the lectin binding characteristics of cytotoxic T lymphocyte (CTL)-derived surface labeled glycoproteins by affinity chromatography of the labeled glycoproteins on a panel of immobilized lectin adsorbents. Evidence is presented for the specific interaction of the CTL-associated glycoprotein T 145 with a lectin derived from Vicia villosa seeds. Conditions are described for the preparation and use of lectin affinity adsorbents for the rapid isolation of T 145 bearing cytotoxic T lymphocytes. Direct proof is given to show that T 145-positive cells arising from a variety of T-cell activations constitute the only subpopulation of cells with ability to perform cell- mediated T-cell cytotoxicity. Specific depletion of the CTLs by adherence to V. villosa adsorbents is shown by their depletion in the nonbound cell fraction and correspondingly enriched recovery in the sugar eluted cell fraction. Specific affinity fractionation of CTLs has occurred in every strain combination tested and irrespective of the actual antigen specificity of the effector cell population.

Full Text

The Full Text of this article is available as a PDF (800.5 KB).

Selected References

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

  1. Andersson L. C., Gahmberg C. G., Kimura A. K., Wigzell H. Activated human T lymphocytes display new surface glycoproteins. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3455–3458. doi: 10.1073/pnas.75.7.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axelsson B., Kimura A., Hammarström S., Wigzell H., Nilsson K., Mellstedt H. Helix pomatia A hemagglutinin: selectivity of binding to lymphocyte surface glycoproteins on T cells and certain B cells. Eur J Immunol. 1978 Nov;8(11):757–764. doi: 10.1002/eji.1830081102. [DOI] [PubMed] [Google Scholar]
  3. Bevan M. J., Cohn M. Cytotoxic effects of antigen- and mitogen-induced T cells on various targets. J Immunol. 1975 Feb;114(2 Pt 1):559–565. [PubMed] [Google Scholar]
  4. Cantor H., Boyse E. A. Functional subclasses of T-lymphocytes bearing different Ly antigens. I. The generation of functionally distinct T-cell subclasses is a differentiative process independent of antigen. J Exp Med. 1975 Jun 1;141(6):1376–1389. doi: 10.1084/jem.141.6.1376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cerottini J. C., Brunner K. T. Cell-mediated cytotoxicity, allograft rejection, and tumor immunity. Adv Immunol. 1974;18:67–132. doi: 10.1016/s0065-2776(08)60308-9. [DOI] [PubMed] [Google Scholar]
  6. Clark W. R. An antigen-specific component of lectin-mediated cytotoxicity. Cell Immunol. 1975 Jun;17(2):505–516. doi: 10.1016/s0008-8749(75)80054-2. [DOI] [PubMed] [Google Scholar]
  7. Cullen S. E., Schwartz B. D. An improved method for isolation of H-2 and Ia alloantigens with immunoprecipitation induced by protein A-bearing staphylococci. J Immunol. 1976 Jul;117(1):136–142. [PubMed] [Google Scholar]
  8. Dawson J. R., Silver J., Sheppard L. B., Amos D. B. The purification of detergent-solubilized HL-A antigens by affinity chromatography with the hemagglutinin from Lens culinaris. J Immunol. 1974 Mar;112(3):1190–1193. [PubMed] [Google Scholar]
  9. Dorval G., Welsh K. I., Wigzell H. Labeled staphylococcal protein A as an immunological probe in the analysis of cell surface markers. Scand J Immunol. 1974;3(4):405–411. doi: 10.1111/j.1365-3083.1974.tb01273.x. [DOI] [PubMed] [Google Scholar]
  10. Golstein P., Smith E. T. Mechanism of T-cell-mediated cytolysis: the lethal hit stage. Contemp Top Immunobiol. 1977;7:273–300. doi: 10.1007/978-1-4684-3054-7_8. [DOI] [PubMed] [Google Scholar]
  11. Hayman M. J., Crumpton M. J. Isolation of glycoproteins from pig lymphocyte plasma membrane using Lens culinaris phytohemagglutinin. Biochem Biophys Res Commun. 1972 May 26;47(4):923–930. doi: 10.1016/0006-291x(72)90581-5. [DOI] [PubMed] [Google Scholar]
  12. Jandinski J., Cantor H., Tadakuma T., Peavy D. L., Pierce C. W. Separation of helper T cells from suppressor T cells expressing different Ly components. I. Polyclonal activation: suppressor and helper activities are inherent properties of distinct T-cell subclasses. J Exp Med. 1976 Jun 1;143(6):1382–1390. doi: 10.1084/jem.143.6.1382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kimura A. K., Wigzell H. Cell surface glycoproteins of murine cytotoxic T lymphocytes. I. T 145, a new cell surface glycoprotein selectively expressed on Ly 1-2+ cytotoxic T lymphocytes. J Exp Med. 1978 May 1;147(5):1418–1434. doi: 10.1084/jem.147.5.1418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kimura A. K., Wigzell H. Cytotoxic T lymphocyte membrane components: an analysis of structures related to function. Contemp Top Mol Immunol. 1977;6:209–244. doi: 10.1007/978-1-4684-2841-4_7. [DOI] [PubMed] [Google Scholar]
  15. Kristiansen T., Sundberg L., Porath J. Studies on blood group substances. II. Coupling of blood group substane A to hydroxyl-containing matrices, including aminoethyl cellulose and agarose. Biochim Biophys Acta. 1969 Jun 17;184(1):93–98. [PubMed] [Google Scholar]
  16. 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]
  17. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  18. Martz E. Mechanism of specific tumor-cell lysis by alloimmune T lymphocytes: resolution and characterization of discrete steps in the cellular interaction. Contemp Top Immunobiol. 1977;7:301–361. doi: 10.1007/978-1-4684-3054-7_9. [DOI] [PubMed] [Google Scholar]
  19. Nabholz M., Vives J., Young H. M., Meo T., Miggiano V., Rijnbeek A., Shreffler D. C. Cell-mediated cell lysis in vitro: genetic control of killer cell production and target specificities in the mouse. Eur J Immunol. 1974 May;4(5):378–387. doi: 10.1002/eji.1830040514. [DOI] [PubMed] [Google Scholar]
  20. Weston P. D., Avrameas S. Proteins coupled to polyacrylamide beads using glutaraldehyde. Biochem Biophys Res Commun. 1971 Dec 17;45(6):1574–1580. doi: 10.1016/0006-291x(71)90200-2. [DOI] [PubMed] [Google Scholar]
  21. Wigzell H. Specific affinity fractionation of lymphocytes using glass or plastic bead columns. Scand J Immunol. 1976 Jun;Suppl 5:23–30. doi: 10.1111/j.1365-3083.1976.tb03853.x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES