Intrathymic Injection for Analysis of T-Cell Progenitor Activity

  • Libuse Jerabek
  • Irving L. Weissman
Protocol
Part of the Methods in Molecular Medicine book series (MIMM, volume 63)

Abstract

Within our field, improvement in fluorescence-activated cell sorting (FACS) and molecular technologies has led to various types of correlative studies that imply the developmental sequence and subsequent emigration of thymic-lymphocyte subsets. Unfortunately, the implied conclusions are often accepted unequivocally by most of the immunology community. In fact, direct demonstration of precursor progeny relationships by specific cell marking within the thymus, or specific delivery of purified cells at a particular stage of isolation back into the thymus, are the only methods that reproducibly identify cell stages and intermediates (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11).

References

  1. 1.
    Weissman, I. L. (1967) Thymus cell migration. J. Exp. Med. 126, 291–304.PubMedCrossRefGoogle Scholar
  2. 2.
    Weissman, I. L. (1973) Thymus cell maturation: studies on the origin of cortisone-resistant thymic lymphocytes. J. Exp. Med. 137, 504–510.PubMedCrossRefGoogle Scholar
  3. 3.
    Scollay, R., Kochen, M., Butcher, E., and Weissman, I. L. (1978) Lyt markers on thymus cell migrants. Nature 276, 79–80.PubMedCrossRefGoogle Scholar
  4. 4.
    Scollay, R., Butcher, E., and Weissman, I. L. (1980) Thymus cell migration. Quantitative aspects of cellular traffic from the thymus to the periphery in mice. Eur. J. Immunol. 10, 210–218.PubMedCrossRefGoogle Scholar
  5. 5.
    Scollay, R., Jacobs, S., Jerabek, L., Butcher, E., and Weissman, I. L. (1980) T cell maturation: thymocyte and thymus migrant subpopulations defined with monoclonal antibodies to MHC region antigens. J. Immunol. 124, 2845–2853.PubMedGoogle Scholar
  6. 6.
    Scollay, R. and Weissman, I. L. (1980) T cell maturation: thymocyte and thymus migrant subpopulations defined with monoclonal antibodies to the antigens Lyt-1, Lyt-2 and ThB. J. Immunol. 124, 2841–2844.PubMedGoogle Scholar
  7. 7.
    Butcher, E. C., Scollay, R. G., and Weissman, I. L. (1980) Direct fluorescent labeling of cells with fluorescein or rhodamine isothiocyanate: II. Potential application to studies of lymphocyte migration and maturation. J. Immunol. Methods 37, 109–121.PubMedCrossRefGoogle Scholar
  8. 8.
    Guidos, C. J., Danska, J. S., Fathman, C. G., and Weissman, I. L. (1990) TCR-mediated negative selection of autoreactive T lymphcyte precursors occurs after commitment to the CD4 or CD8 lineages. J. Exp. Med. 172, 835–845.PubMedCrossRefGoogle Scholar
  9. 9.
    Akashi, K. and Weissman, I. L. (1996) The c-kit positive maturation pathway in mouse thymic T cell development: lineages and selection. Immunity 5, 147–161.PubMedCrossRefGoogle Scholar
  10. 10.
    Akashi, K., Kondo, M., and Weissman, I. L. (1998) Two distinct pathways of positive selection for thymocytes. PNAS 95, 2486–2491.PubMedCrossRefGoogle Scholar
  11. 11.
    Akashi, K., Richie, L. I., Miyamoto, T., Carr, W. H., and Weissman, I. L. B (2000) lymphopoiesis in the thymus. J. Immunol. 164, 5221–5226.PubMedGoogle Scholar
  12. 12.
    Spangrude, G. J., Muller-Sieburg, C. E., Heimfeld, S., and Weissman, I. L. (1988) Two rare populations of mouse Thy-1l1o bone marrow cells repopulate the thymus. J. Exp. Med. 167, 1671–1683.PubMedCrossRefGoogle Scholar
  13. 13.
    Spangrude, G. J., Heimfeld, S., and Weissman, I. L. (1988) Purification and characterization of mouse hematopoietic stem cells. Science 241, 58–62.PubMedCrossRefGoogle Scholar
  14. 14.
    Spangrude, G. J. and Weissman, I. L. (1988) Mature T cells generated from single thymic clones are phenotypically and functionally heterogeneous. J. Immunol. 141, 1877–1890.PubMedGoogle Scholar
  15. 15.
    Kondo, M., Weissman, I. L., and Akashi, K. (1997) Identification of clonogenic common lymphoid progenitor in mouse bone marrow. Cell 91, 661–672.PubMedCrossRefGoogle Scholar
  16. 16.
    Adkins, B., Mueller, C., Okada, C. Y., Reichert, R. A., Weissman, I. L., and Spangrude, G. J. (1987) Early events in T-cell maturation. Annu. Rev. Immunol. 5, 325–365.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Libuse Jerabek
    • 1
  • Irving L. Weissman
    • 1
  1. 1.Stanford University School of MedicineStanford

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