Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1996 Aug 1;184(2):519–530. doi: 10.1084/jem.184.2.519

Regulation of pre-T cell receptor (pT alpha-TCR beta) gene expression during human thymic development

PMCID: PMC2192728  PMID: 8760805

Abstract

In murine T cell development, early thymocytes that productively rearrange the T cell receptor (TCR) beta locus are selected to continue maturation, before TCR alpha expression, by means of a pre-TCR alpha- (pT alpha-) TCR beta heterodimer (pre-TCR). The aim of this study was to identify equivalent stages in human thymocyte development. We show here that variable-diversity-joining region TCR beta rearrangement and the expression of full-length TCR beta transcripts have been initiated in some immature thymocytes at the TCR alpha/beta- CD4+CD8- stage, and become common in a downstream subset of TCR alpha/beta- CD4+CD8+ thymocytes that is highly enriched in large cycling cells. TCR beta chain expression was hardly detected in TCR alpha/beta- CD4+CD8- thymocytes, whereas cytoplasmic TCR beta chain was found in virtually all TCR alpha/beta- CD4+CD8+ blasts. In addition, a TCR beta complex distinct from the mature TCR alpha/beta heterodimer was immunoprecipitated only from the latter subset. cDNA derived from TCR alpha/beta- CD4+CD8+ blasts allowed us to identify and clone the gene encoding the human pT alpha chain, and to examine its expression at different stages of thymocyte development. Our results show that high pT alpha transcription occurs only in CD4+CD8- and CD4+CD8+ TCR alpha/beta- thymocytes, whereas it is weaker in earlier and later stages of development. Based on these results, we propose that the transition from TCR alpha/beta- CD4+CD8- to TCR alpha/beta- CD4+CD8+ thymocytes represents a critical developmental stage at which the successful expression of TCR beta promotes the clonal expansion and further maturation of human thymocytes, independent of TCR alpha.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

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

  1. Anderson S. J., Abraham K. M., Nakayama T., Singer A., Perlmutter R. M. Inhibition of T-cell receptor beta-chain gene rearrangement by overexpression of the non-receptor protein tyrosine kinase p56lck. EMBO J. 1992 Dec;11(13):4877–4886. doi: 10.1002/j.1460-2075.1992.tb05594.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bairoch A., Bucher P., Hofmann K. The PROSITE database, its status in 1995. Nucleic Acids Res. 1996 Jan 1;24(1):189–196. doi: 10.1093/nar/24.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brenner M. B., McLean J., Scheft H., Warnke R. A., Jones N., Strominger J. L. Characterization and expression of the human alpha beta T cell receptor by using a framework monoclonal antibody. J Immunol. 1987 Mar 1;138(5):1502–1509. [PubMed] [Google Scholar]
  4. Broeren C. P., Verjans G. M., Van Eden W., Kusters J. G., Lenstra J. A., Logtenberg T. Conserved nucleotide sequences at the 5' end of T cell receptor variable genes facilitate polymerase chain reaction amplification. Eur J Immunol. 1991 Mar;21(3):569–575. doi: 10.1002/eji.1830210306. [DOI] [PubMed] [Google Scholar]
  5. Bruno L., Rocha B., Rolink A., von Boehmer H., Rodewald H. R. Intra- and extra-thymic expression of the pre-T cell receptor alpha gene. Eur J Immunol. 1995 Jul;25(7):1877–1882. doi: 10.1002/eji.1830250713. [DOI] [PubMed] [Google Scholar]
  6. Choi Y. W., Kotzin B., Herron L., Callahan J., Marrack P., Kappler J. Interaction of Staphylococcus aureus toxin "superantigens" with human T cells. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8941–8945. doi: 10.1073/pnas.86.22.8941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Del Porto P., Bruno L., Mattei M. G., von Boehmer H., Saint-Ruf C. Cloning and comparative analysis of the human pre-T-cell receptor alpha-chain gene. Proc Natl Acad Sci U S A. 1995 Dec 19;92(26):12105–12109. doi: 10.1073/pnas.92.26.12105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dudley E. C., Petrie H. T., Shah L. M., Owen M. J., Hayday A. C. T cell receptor beta chain gene rearrangement and selection during thymocyte development in adult mice. Immunity. 1994 May;1(2):83–93. doi: 10.1016/1074-7613(94)90102-3. [DOI] [PubMed] [Google Scholar]
  9. Ernst B., Surh C. D., Sprent J. Thymic selection and cell division. J Exp Med. 1995 Oct 1;182(4):961–971. doi: 10.1084/jem.182.4.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fehling H. J., Krotkova A., Saint-Ruf C., von Boehmer H. Crucial role of the pre-T-cell receptor alpha gene in development of alpha beta but not gamma delta T cells. Nature. 1995 Jun 29;375(6534):795–798. doi: 10.1038/375795a0. [DOI] [PubMed] [Google Scholar]
  11. Galy A., Verma S., Bárcena A., Spits H. Precursors of CD3+CD4+CD8+ cells in the human thymus are defined by expression of CD34. Delineation of early events in human thymic development. J Exp Med. 1993 Aug 1;178(2):391–401. doi: 10.1084/jem.178.2.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Godfrey D. I., Kennedy J., Mombaerts P., Tonegawa S., Zlotnik A. Onset of TCR-beta gene rearrangement and role of TCR-beta expression during CD3-CD4-CD8- thymocyte differentiation. J Immunol. 1994 May 15;152(10):4783–4792. [PubMed] [Google Scholar]
  13. Godfrey D. I., Zlotnik A. Control points in early T-cell development. Immunol Today. 1993 Nov;14(11):547–553. doi: 10.1016/0167-5699(93)90186-O. [DOI] [PubMed] [Google Scholar]
  14. Groettrup M., Ungewiss K., Azogui O., Palacios R., Owen M. J., Hayday A. C., von Boehmer H. A novel disulfide-linked heterodimer on pre-T cells consists of the T cell receptor beta chain and a 33 kd glycoprotein. Cell. 1993 Oct 22;75(2):283–294. doi: 10.1016/0092-8674(93)80070-u. [DOI] [PubMed] [Google Scholar]
  15. Hori T., Cupp J., Wrighton N., Lee F., Spits H. Identification of a novel human thymocyte subset with a phenotype of CD3- CD4+ CD8 alpha + beta-1. Possible progeny of the CD3- CD4- CD8- subset. J Immunol. 1991 Jun 15;146(12):4078–4084. [PubMed] [Google Scholar]
  16. Koning F., Lew A. M., Maloy W. L., Valas R., Coligan J. E. The biosynthesis and assembly of T cell receptor alpha- and beta-chains with the CD3 complex. J Immunol. 1988 May 1;140(9):3126–3134. [PubMed] [Google Scholar]
  17. Kraft D. L., Weissman I. L., Waller E. K. Differentiation of CD3-4-8- human fetal thymocytes in vivo: characterization of a CD3-4+8- intermediate. J Exp Med. 1993 Jul 1;178(1):265–277. doi: 10.1084/jem.178.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ktorza S., Sarun S., Rieux-Laucat F., de Villartay J. P., Debré P., Schmitt C. CD34-positive early human thymocytes: T cell receptor and cytokine receptor gene expression. Eur J Immunol. 1995 Sep;25(9):2471–2478. doi: 10.1002/eji.1830250910. [DOI] [PubMed] [Google Scholar]
  19. Levelt C. N., Carsetti R., Eichmann K. Regulation of thymocyte development through CD3. II. Expression of T cell receptor beta CD3 epsilon and maturation to the CD4+8+ stage are highly correlated in individual thymocytes. J Exp Med. 1993 Dec 1;178(6):1867–1875. doi: 10.1084/jem.178.6.1867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Levelt C. N., Eichmann K. Receptors and signals in early thymic selection. Immunity. 1995 Dec;3(6):667–672. doi: 10.1016/1074-7613(95)90056-x. [DOI] [PubMed] [Google Scholar]
  21. Malissen M., Gillet A., Ardouin L., Bouvier G., Trucy J., Ferrier P., Vivier E., Malissen B. Altered T cell development in mice with a targeted mutation of the CD3-epsilon gene. EMBO J. 1995 Oct 2;14(19):4641–4653. doi: 10.1002/j.1460-2075.1995.tb00146.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McMichael A. J., Pilch J. R., Galfré G., Mason D. Y., Fabre J. W., Milstein C. A human thymocyte antigen defined by a hybrid myeloma monoclonal antibody. Eur J Immunol. 1979 Mar;9(3):205–210. doi: 10.1002/eji.1830090307. [DOI] [PubMed] [Google Scholar]
  23. Melchers F., Rolink A., Grawunder U., Winkler T. H., Karasuyama H., Ghia P., Andersson J. Positive and negative selection events during B lymphopoiesis. Curr Opin Immunol. 1995 Apr;7(2):214–227. doi: 10.1016/0952-7915(95)80006-9. [DOI] [PubMed] [Google Scholar]
  24. Mombaerts P., Clarke A. R., Rudnicki M. A., Iacomini J., Itohara S., Lafaille J. J., Wang L., Ichikawa Y., Jaenisch R., Hooper M. L. Mutations in T-cell antigen receptor genes alpha and beta block thymocyte development at different stages. Nature. 1992 Nov 19;360(6401):225–231. doi: 10.1038/360225a0. [DOI] [PubMed] [Google Scholar]
  25. Márquez C., Trigueros C., Fernández E., Toribio M. L. The development of T and non-T cell lineages from CD34+ human thymic precursors can be traced by the differential expression of CD44. J Exp Med. 1995 Feb 1;181(2):475–483. doi: 10.1084/jem.181.2.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nikolić-Zugić J., Moore M. W., Bevan M. J. Characterization of the subset of immature thymocytes which can undergo rapid in vitro differentiation. Eur J Immunol. 1989 Apr;19(4):649–653. doi: 10.1002/eji.1830190412. [DOI] [PubMed] [Google Scholar]
  27. Okada A., Mendelsohn M., Alt F. Differential activation of transcription versus recombination of transgenic T cell receptor beta variable region gene segments in B and T lineage cells. J Exp Med. 1994 Jul 1;180(1):261–272. doi: 10.1084/jem.180.1.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Péault B., Weissman I. L., Baum C., McCune J. M., Tsukamoto A. Lymphoid reconstitution of the human fetal thymus in SCID mice with CD34+ precursor cells. J Exp Med. 1991 Nov 1;174(5):1283–1286. doi: 10.1084/jem.174.5.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Raulet D. H., Garman R. D., Saito H., Tonegawa S. Developmental regulation of T-cell receptor gene expression. Nature. 1985 Mar 7;314(6006):103–107. doi: 10.1038/314103a0. [DOI] [PubMed] [Google Scholar]
  30. Royer H. D., Ramarli D., Acuto O., Campen T. J., Reinherz E. L. Genes encoding the T-cell receptor alpha and beta subunits are transcribed in an ordered manner during intrathymic ontogeny. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5510–5514. doi: 10.1073/pnas.82.16.5510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Saint-Ruf C., Ungewiss K., Groettrup M., Bruno L., Fehling H. J., von Boehmer H. Analysis and expression of a cloned pre-T cell receptor gene. Science. 1994 Nov 18;266(5188):1208–1212. doi: 10.1126/science.7973703. [DOI] [PubMed] [Google Scholar]
  32. Shinkai Y., Koyasu S., Nakayama K., Murphy K. M., Loh D. Y., Reinherz E. L., Alt F. W. Restoration of T cell development in RAG-2-deficient mice by functional TCR transgenes. Science. 1993 Feb 5;259(5096):822–825. doi: 10.1126/science.8430336. [DOI] [PubMed] [Google Scholar]
  33. Shortman K., Vremec D., Egerton M. The kinetics of T cell antigen receptor expression by subgroups of CD4+8+ thymocytes: delineation of CD4+8+3(2+) thymocytes as post-selection intermediates leading to mature T cells. J Exp Med. 1991 Feb 1;173(2):323–332. doi: 10.1084/jem.173.2.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Spits H. Early stages in human and mouse T-cell development. Curr Opin Immunol. 1994 Apr;6(2):212–221. doi: 10.1016/0952-7915(94)90094-9. [DOI] [PubMed] [Google Scholar]
  35. Sánchez M. J., Gutiérrez-Ramos J. C., Fernández E., Leonardo E., Lozano J., Martínez C., Toribio M. L. Putative prethymic T cell precursors within the early human embryonic liver: a molecular and functional analysis. J Exp Med. 1993 Jan 1;177(1):19–33. doi: 10.1084/jem.177.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Toribio M. L., Gutiérrez-Ramos J. C., Pezzi L., Marcos M. A., Martínez C. Interleukin-2-dependent autocrine proliferation in T-cell development. Nature. 1989 Nov 2;342(6245):82–85. doi: 10.1038/342082a0. [DOI] [PubMed] [Google Scholar]
  37. Toyonaga B., Yoshikai Y., Vadasz V., Chin B., Mak T. W. Organization and sequences of the diversity, joining, and constant region genes of the human T-cell receptor beta chain. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8624–8628. doi: 10.1073/pnas.82.24.8624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Turka L. A., Schatz D. G., Oettinger M. A., Chun J. J., Gorka C., Lee K., McCormack W. T., Thompson C. B. Thymocyte expression of RAG-1 and RAG-2: termination by T cell receptor cross-linking. Science. 1991 Aug 16;253(5021):778–781. doi: 10.1126/science.1831564. [DOI] [PubMed] [Google Scholar]
  39. Wilson A., Held W., MacDonald H. R. Two waves of recombinase gene expression in developing thymocytes. J Exp Med. 1994 Apr 1;179(4):1355–1360. doi: 10.1084/jem.179.4.1355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yanagi Y., Chan A., Chin B., Minden M., Mak T. W. Analysis of cDNA clones specific for human T cells and the alpha and beta chains of the T-cell receptor heterodimer from a human T-cell line. Proc Natl Acad Sci U S A. 1985 May;82(10):3430–3434. doi: 10.1073/pnas.82.10.3430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Yoshikai Y., Anatoniou D., Clark S. P., Yanagi Y., Sangster R., Van den Elsen P., Terhorst C., Mak T. W. Sequence and expression of transcripts of the human T-cell receptor beta-chain genes. Nature. 1984 Dec 6;312(5994):521–524. doi: 10.1038/312521a0. [DOI] [PubMed] [Google Scholar]
  42. von Boehmer H. Developmental biology of T cells in T cell-receptor transgenic mice. Annu Rev Immunol. 1990;8:531–556. doi: 10.1146/annurev.iy.08.040190.002531. [DOI] [PubMed] [Google Scholar]

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

RESOURCES