Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1974 Aug 1;62(2):366–377. doi: 10.1083/jcb.62.2.366

THE KINETICS OF CELLULAR COMMITMENT DURING STIMULATION OF LYMPHOCYTES BY LECTINS

Gary R Gunther 1, John L Wang 1, Gerald M Edelman 1
PMCID: PMC2109389  PMID: 4426912

Abstract

The kinetics of cellular commitment in the stimulation of lymphocytes by concanavalin A (Con A) has been analyzed by measurement of DNA synthesis, autoradiography, and histologic staining techniques. If the competitive inhibitor α-methyl-D-mannoside (αMM) is introduced into cultures of mouse spleen cells at various times after the addition of Con A, there is a gradual decrease in its capacity to inhibit the lectin-stimulated incorporation of [3H]thymidine. Addition of the saccharide 20 h after exposure of the cells to Con A had no effect on the level of the cellular response to the lectin. With increasing periods of contact with Con A, the percentage of blast cells and the percentage of [3H]thymidine-labeled blast cells increased in parallel with the total radioactive thymidine incorporated while the average number of autoradiographic grains per labeled blast cell remained relatively constant. These observations suggest that the rising level of [3H]thymidine incorporation results from an increase in the number of cells that respond to lectin stimulation and become refractory to inhibition with αMM. Once such cells become committed, they synthesize DNA at a rate independent of the length of exposure to the lectin. The combined results indicate that mouse splenic lymphocytes are heterogeneous in their capacities to respond to Con A and that different cells require different induction periods to be stimulated.

Full Text

The Full Text of this article is available as a PDF (745.9 KB).

Selected References

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

  1. Byrt P., Ada G. L. An in vitro reaction between labelled flagellin or haemocyanin and lymphocyte-like cells from normal animals. Immunology. 1969 Oct;17(4):503–516. [PMC free article] [PubMed] [Google Scholar]
  2. COOPER E. H., BARKHAN P., HALE A. J. Observations on the proliferation of human leucocytes cultured with phytohaemagglutinin. Br J Haematol. 1963 Jan;9:101–111. doi: 10.1111/j.1365-2141.1963.tb05446.x. [DOI] [PubMed] [Google Scholar]
  3. Cunningham B. A., Wang J. L., Pflumm M. N., Edelman G. M. Isolation and proteolytic cleavage of the intact subunit of concanavalin A. Biochemistry. 1972 Aug 15;11(17):3233–3239. doi: 10.1021/bi00767a016. [DOI] [PubMed] [Google Scholar]
  4. Edelman G. M., Cunningham B. A., Reeke G. N., Jr, Becker J. W., Waxdal M. J., Wang J. L. The covalent and three-dimensional structure of concanavalin A. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2580–2584. doi: 10.1073/pnas.69.9.2580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. GOLDSTEIN I. J., HOLLERMAN C. E., SMITH E. E. PROTEIN-CARBOHYDRATE INTERACTION. II. INHIBITION STUDIES ON THE INTERACTION OF CONCANAVALIN A WITH POLYSACCHARIDES. Biochemistry. 1965 May;4:876–883. doi: 10.1021/bi00881a013. [DOI] [PubMed] [Google Scholar]
  6. Gunther G. R., Wang J. L., Yahara I., Cunningham B. A., Edelman G. M. Concanavalin A derivatives with altered biological activities. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1012–1016. doi: 10.1073/pnas.70.4.1012. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HALE A. J., COOPER E. H., MILTON J. D. STUDIES OF THE INCORPORATION OF PYRIMIDINES INTO DNA IN SINGLE LEUKAEMIC AND OTHER PROLIFERATING LEUCOCYTES. Br J Haematol. 1965 Mar;11:144–161. doi: 10.1111/j.1365-2141.1965.tb06572.x. [DOI] [PubMed] [Google Scholar]
  8. Jones G. Lymphocyte activation. 3. The prolonged requirement for mitogen in phytohaemagglutinin- and concanavalin A-stimulated cultures. J Immunol. 1973 May;110(5):1262–1267. [PubMed] [Google Scholar]
  9. Jones G. The number of reactive cells in mouse lymphocyte cultures stimulated by phytohemagglutinin, concanavalin A or histocompatibility antigen. J Immunol. 1973 Sep;111(3):914–920. [PubMed] [Google Scholar]
  10. Kay J. E. The role of the stimulant in the activation of lymphocytes by PHA. Exp Cell Res. 1969 Nov;58(1):185–188. doi: 10.1016/0014-4827(69)90134-7. [DOI] [PubMed] [Google Scholar]
  11. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  12. Mishell R. I., Dutton R. W. Immunization of dissociated spleen cell cultures from normal mice. J Exp Med. 1967 Sep 1;126(3):423–442. doi: 10.1084/jem.126.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Novogrodsky A., Katchalski E. Lymphocyte transformation induced by concanavalin A and its reversion by methyl-alpha-D-mannopyranoside. Biochim Biophys Acta. 1971 Jan 28;228(2):579–583. doi: 10.1016/0005-2787(71)90064-5. [DOI] [PubMed] [Google Scholar]
  14. Powell A. E., Leon M. A. Reversible interaction of human lymphocytes with the mitogen concanavalin A. Exp Cell Res. 1970 Oct;62(2):315–325. doi: 10.1016/0014-4827(70)90560-4. [DOI] [PubMed] [Google Scholar]
  15. ROGERS A. W. A simple photometric device for the quantitation of silver grains in autoradiographs of tissue sections. Exp Cell Res. 1961 Aug;24:228–239. doi: 10.1016/0014-4827(61)90425-6. [DOI] [PubMed] [Google Scholar]
  16. Smith J. A., Martin L. Do cells cycle? Proc Natl Acad Sci U S A. 1973 Apr;70(4):1263–1267. doi: 10.1073/pnas.70.4.1263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Temin H. M. Stimulation by serum of multiplication of stationary chicken cells. J Cell Physiol. 1971 Oct;78(2):161–170. doi: 10.1002/jcp.1040780202. [DOI] [PubMed] [Google Scholar]
  18. Woodruff M. F., Reid B., James K. Effect of antilymphocytic antibody and antibody fragments on human lymphocytes in vitro. Nature. 1967 Aug 5;215(5101):591–594. doi: 10.1038/215591a0. [DOI] [PubMed] [Google Scholar]
  19. Yahara I., Edelman G. M. Restriction of the mobility of lymphocyte immunoglobulin receptors by concanavalin A. Proc Natl Acad Sci U S A. 1972 Mar;69(3):608–612. doi: 10.1073/pnas.69.3.608. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES