Examination of thymic positive and negative selection by flow cytometry

Journal of Visualized Experiments

Volumn , Issue 68, 2012, Pages

  Hu, Qian ^a   Nicol, Stephanie A ^a   Suen, Alexander Y W ^a   Baldwin, Troy A ^a  

^a UNIVERSITY OF ALBERTA   (Canada)

Author keywords

Anatomy; Autoimmunity; Cellular biology; Flow cytometry; Immunology; Issue 68; Medicine; Negative selection; Physiology; Positive selection; T cell; Thymus

Indexed keywords

EID: 84868295253     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/4269     Document Type: Article

References (32)

1. Liston, A., Lesage, S., Wilson, J., Peltonen, L., & Goodnow, C.C. Aire regulates negative selection of organ-specific T cells. Nat. Immunol. 4, 350-354 (2003).
2. Liston, A., et al. Gene dosage--limiting role of Aire in thymic expression, clonal deletion, and organ-specific autoimmunity. J. Exp. Med. 200, 1015-1026 (2004).
3. Hogquist, K.A., Baldwin, T.A., & Jameson, S.C. Central tolerance: learning self-control in the thymus. Nat. Rev. Immunol. 5, 772-782 (2005).
4. Liston, A., Enders, A., & Siggs, O.M. Unravelling the association of partial T-cell immunodeficiency and immune dysregulation. Nat. Rev. Immunol. 8, 545-558 (2008).
5. Starr, T.K., Jameson, S.C., & Hogquist, K.A. Positive and negative selection of T cells. Annu. Rev. Immunol. 21, 139-176 (2003).
6. McCaughtry, T.M. & Hogquist, K.A. Central tolerance: what have we learned from mice? Seminars in immunopathology. 30, 399-409 (2008).
7. Zhan, Y., et al. Without peripheral interference, thymic deletion is mediated in a cohort of double-positive cells without classical activation. Proceedings of the National Academy of Sciences of the United States of America. 100, 1197-1202 (2003).
8. Brewer, J.A., Kanagawa, O., Sleckman, B.P., & Muglia, L.J. Thymocyte apoptosis induced by T cell activation is mediated by glucocorticoids in vivo. J. Immunol. 169, 1837-1843 (2002).
9. Martin, S. & Bevan, M.J. Antigen-specific and nonspecific deletion of immature cortical thymocytes caused by antigen injection. European journal of immunology. 27, 2726-2736 (1997).
10. Baldwin, T.A., Sandau, M.M., Jameson, S.C., & Hogquist, K.A. The timing of TCR alpha expression critically influences T cell development and selection. J. Exp. Med. 202, 111-121 (2005).
11. Tung, J.W., et al. Modern flow cytometry: a practical approach. Clinics in laboratory medicine. 27, 453-468, v (2007).
12. Aliahmad, P. & Kaye, J. Development of all CD4 T lineages requires nuclear factor TOX. J. Exp. Med. 205, 245-256 (2008).
13. Kastner, P., et al. Bcl11b represses a mature T-cell gene expression program in immature CD4(+)CD8(+) thymocytes. Eur. J. Immunol. 40, 2143-2154 (2010).
14. Albu, D.I., et al. BCL11B is required for positive selection and survival of double-positive thymocytes. J. Exp. Med. 204, 3003-3015 (2007).
15. Van De Wiele, C.J., et al. Thymocytes between the beta-selection and positive selection checkpoints are nonresponsive to IL-7 as assessed by STAT-5 phosphorylation. J. Immunol. 172, 4235-4244 (2004).
16. Ueno, T., et al. CCR7 signals are essential for cortex-medulla migration of developing thymocytes. J. Exp. Med. 200, 493-505 (2004).
17. Saini, M., et al. Regulation of Zap70 expression during thymocyte development enables temporal separation of CD4 and CD8 repertoire selection at different signaling thresholds. Science signaling. 3, ra23 (2010).
18. Hu, Q., Sader, A., Parkman, J.C., & Baldwin, T.A. Bim-mediated apoptosis is not necessary for thymic negative selection to ubiquitous self-antigens. J. Immunol. 183, 7761-7767 (2009).
19. Kisielow, P., Bluthmann, H., Staerz, U.D., Steinmetz, M., & von Boehmer, H. Tolerance in T-cell-receptor transgenic mice involves deletion of nonmature CD4+8+ thymocytes. Nature. 333, 742-746 (1988).
20. McCaughtry, T.M., Baldwin, T.A., Wilken, M.S., & Hogquist, K.A. Clonal deletion of thymocytes can occur in the cortex with no involvement of the medulla. J. Exp. Med. 205, 2575-2584 (2008).
21. Derbinski, J., Schulte, A., Kyewski, B., & Klein, L. Promiscuous gene expression in medullary thymic epithelial cells mirrors the peripheral self. Nat. Immunol. 2, 1032-1039 (2001).
22. Anderson, M.S., et al. Projection of an immunological self shadow within the thymus by the aire protein. Science. 298, 1395-1401 (2002).
23. Kurts, C., et al. Constitutive class I-restricted exogenous presentation of self antigens in vivo. J. Exp. Med. 184, 923-930 (1996).
24. Nitta, T., Nitta, S., Lei, Y., Lipp, M., & Takahama, Y. CCR7-mediated migration of developing thymocytes to the medulla is essential for negative selection to tissue-restricted antigens. Proceedings of the National Academy of Sciences of the United States of America. 106, 17129-17133 (2009).
25. Bouneaud, C., Kourilsky, P., & Bousso, P. Impact of negative selection on the T cell repertoire reactive to a self-peptide: a large fraction of T cell clones escapes clonal deletion. Immunity. 13, 829-840 (2000).
26. Gallegos, A.M. & Bevan, M.J. Central tolerance to tissue-specific antigens mediated by direct and indirect antigen presentation. J. Exp. Med. 200, 1039-1049 (2004).
27. Moon, J.J., et al. Naive CD4(+) T cell frequency varies for different epitopes and predicts repertoire diversity and response magnitude. Immunity. 27, 203-213 (2007).
28. Bouillet, P., et al. BH3-only Bcl-2 family member Bim is required for apoptosis of autoreactive thymocytes. Nature. 415, 922-926 (2002).
29. Suen, A.Y. & Baldwin, T.A. Proapoptotic protein Bim is differentially required during thymic clonal deletion to ubiquitous versus tissue-restricted antigens. Proceedings of the National Academy of Sciences of the United States of America. (2012).
30. Calnan, B.J., Szychowski, S., Chan, F.K., Cado, D., & Winoto, A. A role for the orphan steroid receptor Nur77 in apoptosis accompanying antigen-induced negative selection. Immunity. 3, 273-282 (1995).
31. Zhou, T., et al. Inhibition of Nur77/Nurr1 leads to inefficient clonal deletion of self-reactive T cells. J. Exp. Med. 183, 1879-1892 (1996).
32. Baldwin, T.A. & Hogquist, K.A. Transcriptional analysis of clonal deletion in vivo. J. Immunol. 179, 837-844 (2007).