Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Jul 1;99(1 Pt 1):350–355. doi: 10.1083/jcb.99.1.350

Spectrin immunofluorescence distinguishes a population of naturally capped lymphocytes in situ

PMCID: PMC2275612  PMID: 6376522

Abstract

Immunofluorescence analysis of mammalian lymphocytes using antiserum directed against chicken erythrocyte alpha-spectrin revealed a lymphocyte population in which spectrin antigen was arranged in the form of a discrete cap (hereafter referred to as capped lymphocytes). This subset could be easily distinguished from other lymphocytes in which the spectrin antigen was diffusely distributed near the plasma membrane (noncapped lymphocytes). The subset of capped lymphocytes could be visualized in situ and in isolated cells in the absence of added ligand. Using frozen sections of lymphoid organs that were fixed in formaldehyde prior to the immunofluorescence procedure, capped lymphocytes were found in characteristic locations depending on the tissue examined. In the thymus, the major population of medullary lymphocytes were capped whereas cortical lymphocytes were mostly noncapped. In Peyer's patches, capped lymphocytes were interspersed with noncapped lymphocytes throughout the tissue. In the spleen, capped lymphocytes were concentrated in the periarterial lymphoid sheath of the white pulp and in lymph nodes they were found predominantly in the paracortical and cortical regions. Capped lymphocytes were not visible in the thymus until just before birth and did not appear in the spleen until 3 d after birth. When lymphocytes were isolated from lymphoid organs, fixed in formaldehyde and prepared for immunofluorescence, capped and noncapped lymphocytes were still identifiable and present in the same relative proportions as seen in situ. Results identical to those described above are obtained using antisera directed against guinea pig fodrin. Natural capping of proteins previously shown to co- migrate with a variety of cell surface macromolecules after cross- linking may be a new means of identifying various stages of lymphocyte activation or differentiation.

Full Text

The Full Text of this article is available as a PDF (845.3 KB).

Selected References

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

  1. Bennett V., Davis J., Fowler W. E. Brain spectrin, a membrane-associated protein related in structure and function to erythrocyte spectrin. Nature. 1982 Sep 9;299(5879):126–131. doi: 10.1038/299126a0. [DOI] [PubMed] [Google Scholar]
  2. Bennett V., Stenbuck P. J. The membrane attachment protein for spectrin is associated with band 3 in human erythrocyte membranes. Nature. 1979 Aug 9;280(5722):468–473. doi: 10.1038/280468a0. [DOI] [PubMed] [Google Scholar]
  3. Bennett V. The molecular basis for membrane - cytoskeleton association in human erythrocytes. J Cell Biochem. 1982;18(1):49–65. doi: 10.1002/jcb.1982.240180106. [DOI] [PubMed] [Google Scholar]
  4. Berlin R. D., Oliver J. M. The movement of bound ligands over cell surfaces. J Theor Biol. 1982 Nov 7;99(1):69–80. doi: 10.1016/0022-5193(82)90389-7. [DOI] [PubMed] [Google Scholar]
  5. Branton D., Cohen C. M., Tyler J. Interaction of cytoskeletal proteins on the human erythrocyte membrane. Cell. 1981 Apr;24(1):24–32. doi: 10.1016/0092-8674(81)90497-9. [DOI] [PubMed] [Google Scholar]
  6. Braun J., Fujiwara K., Pollard T. D., Unanue E. R. Two distinct mechanisms for redistribution of lymphocyte surface macromolecules. I. Relationship to cytoplasmic myosin. J Cell Biol. 1978 Nov;79(2 Pt 1):409–418. doi: 10.1083/jcb.79.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burridge K. Direct identification of specific glycoproteins and antigens in sodium dodecyl sulfate gels. Methods Enzymol. 1978;50:54–64. doi: 10.1016/0076-6879(78)50007-4. [DOI] [PubMed] [Google Scholar]
  8. Burridge K., Kelly T., Mangeat P. Nonerythrocyte spectrins: actin-membrane attachment proteins occurring in many cell types. J Cell Biol. 1982 Nov;95(2 Pt 1):478–486. doi: 10.1083/jcb.95.2.478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cantor H., Weissman I. Development and function of subpopulations of thymocytes and T lymphocytes. Prog Allergy. 1976;20:1–64. [PubMed] [Google Scholar]
  10. Cohen C. M. The molecular organization of the red cell membrane skeleton. Semin Hematol. 1983 Jul;20(3):141–158. [PubMed] [Google Scholar]
  11. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  12. Geiger B. Membrane-cytoskeleton interaction. Biochim Biophys Acta. 1983 Aug 11;737(3-4):305–341. doi: 10.1016/0304-4157(83)90005-9. [DOI] [PubMed] [Google Scholar]
  13. Glenney J. R., Jr, Glenney P., Osborn M., Weber K. An F-actin- and calmodulin-binding protein from isolated intestinal brush borders has a morphology related to spectrin. Cell. 1982 Apr;28(4):843–854. doi: 10.1016/0092-8674(82)90063-0. [DOI] [PubMed] [Google Scholar]
  14. Glenney J. R., Jr, Glenney P., Weber K. Erythroid spectrin, brain fodrin, and intestinal brush border proteins (TW-260/240) are related molecules containing a common calmodulin-binding subunit bound to a variant cell type-specific subunit. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4002–4005. doi: 10.1073/pnas.79.13.4002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Glenney J. R., Jr, Glenney P., Weber K. F-actin-binding and cross-linking properties of porcine brain fodrin, a spectrin-related molecule. J Biol Chem. 1982 Aug 25;257(16):9781–9787. [PubMed] [Google Scholar]
  16. Goodman S. R., Shiffer K. The spectrin membrane skeleton of normal and abnormal human erythrocytes: a review. Am J Physiol. 1983 Mar;244(3):C121–C141. doi: 10.1152/ajpcell.1983.244.3.C121. [DOI] [PubMed] [Google Scholar]
  17. Granger B. L., Lazarides E. Synemin: a new high molecular weight protein associated with desmin and vimentin filaments in muscle. Cell. 1980 Dec;22(3):727–738. doi: 10.1016/0092-8674(80)90549-8. [DOI] [PubMed] [Google Scholar]
  18. Granger B. L., Repasky E. A., Lazarides E. Synemin and vimentin are components of intermediate filaments in avian erythrocytes. J Cell Biol. 1982 Feb;92(2):299–312. doi: 10.1083/jcb.92.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hubbard B. D., Lazarides E. Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments. J Cell Biol. 1979 Jan;80(1):166–182. doi: 10.1083/jcb.80.1.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Levine J., Willard M. Fodrin: axonally transported polypeptides associated with the internal periphery of many cells. J Cell Biol. 1981 Sep;90(3):631–642. doi: 10.1083/jcb.90.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Levine J., Willard M. Redistribution of fodrin (a component of the cortical cytoplasm) accompanying capping of cell surface molecules. Proc Natl Acad Sci U S A. 1983 Jan;80(1):191–195. doi: 10.1073/pnas.80.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Marchesi V. T. The red cell membrane skeleton: recent progress. Blood. 1983 Jan;61(1):1–11. [PubMed] [Google Scholar]
  24. Nelson W. J., Colaço C. A., Lazarides E. Involvement of spectrin in cell-surface receptor capping in lymphocytes. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1626–1630. doi: 10.1073/pnas.80.6.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nicolson G. L., Painter R. G. Anionic sites of human erythrocyte membranes. II. Antispectrin-induced transmembrane aggregation of the binding sites for positively charged colloidal particles. J Cell Biol. 1973 Nov;59(2 Pt 1):395–406. doi: 10.1083/jcb.59.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Repasky E. A., Granger B. L., Lazarides E. Widespread occurrence of avian spectrin in nonerythroid cells. Cell. 1982 Jul;29(3):821–833. doi: 10.1016/0092-8674(82)90444-5. [DOI] [PubMed] [Google Scholar]
  27. Tokuyasu K. T., Schekman R., Singer S. J. Domains of receptor mobility and endocytosis in the membranes of neonatal human erythrocytes and reticulocytes are deficient in spectrin. J Cell Biol. 1979 Feb;80(2):481–486. doi: 10.1083/jcb.80.2.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zweig S. E., Tokuyasu K. T., Singer S. J. Member-associated changes during erythropoiesis. On the mechanism of maturation of reticulocytes to erythrocytes. J Supramol Struct Cell Biochem. 1981;17(2):163–181. doi: 10.1002/jsscb.380170207. [DOI] [PubMed] [Google Scholar]

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

RESOURCES