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. 1995 Dec;15(12):7022–7031. doi: 10.1128/mcb.15.12.7022

A delta T-cell receptor deleting element transgenic reporter construct is rearranged in alpha beta but not gamma delta T-cell lineages.

J Shutter 1, J A Cain 1, S Ledbetter 1, M D Rogers 1, R D Hockett Jr 1
PMCID: PMC230957  PMID: 8524269

Abstract

T cells can be divided into two groups on the basis of the expression of either alpha beta or gamma delta T-cell receptors (TCRs). Because the TCR delta chain locus lies within the larger TCR alpha chain locus, control of the utilization of these two receptors is important in T-cell development, specifically for determination of T-cell type: rearrangement of the alpha locus results in deletion of the delta coding segments and commitment to the alpha beta lineage. In the developing thymus, a relative site-specific recombination occurs by which the TCR delta chain gene segments are deleted. This deletion removes all D delta, J delta, and C delta genes and occurs on both alleles. This delta deletional mechanism is evolutionarily conserved between mice and humans. Transgenic mice which contain the human delta deleting elements and as much internal TCR delta chain coding sequence as possible without allowing the formation of a complete delta chain gene were developed. Several transgenic lines showing recombinations between deleting elements within the transgene were developed. These lines demonstrate that utilization of the delta deleting elements occurs in alpha beta T cells of the spleen and thymus. These recombinations are rare in the gamma delta population, indicating that the machinery for utilization of delta deleting elements is functional in alpha beta T cells but absent in gamma delta T cells. Furthermore, a discrete population of early thymocytes containing delta deleting element recombinations but not V alpha-to-J alpha rearrangements has been identified. These data are consistent with a model in which delta deletion contributes to the implementation of a signal by which the TCR alpha chain locus is rearranged and expressed and thus becomes an alpha beta T cell.

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Selected References

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  1. Allison J. P., McIntyre B. W., Bloch D. Tumor-specific antigen of murine T-lymphoma defined with monoclonal antibody. J Immunol. 1982 Nov;129(5):2293–2300. [PubMed] [Google Scholar]
  2. Allison J. P., Raulet D. H. The immunobiology of gamma delta+ T cells. Semin Immunol. 1990 Jan;2(1):59–65. [PubMed] [Google Scholar]
  3. Barth R. K., Kim B. S., Lan N. C., Hunkapiller T., Sobieck N., Winoto A., Gershenfeld H., Okada C., Hansburg D., Weissman I. L. The murine T-cell receptor uses a limited repertoire of expressed V beta gene segments. Nature. 1985 Aug 8;316(6028):517–523. doi: 10.1038/316517a0. [DOI] [PubMed] [Google Scholar]
  4. Born W. K., O'Brien R. L., Modlin R. L. Antigen specificity of gamma delta T lymphocytes. FASEB J. 1991 Sep;5(12):2699–2705. doi: 10.1096/fasebj.5.12.1717333. [DOI] [PubMed] [Google Scholar]
  5. Brenner M. B., McLean J., Dialynas D. P., Strominger J. L., Smith J. A., Owen F. L., Seidman J. G., Ip S., Rosen F., Krangel M. S. Identification of a putative second T-cell receptor. Nature. 1986 Jul 10;322(6075):145–149. doi: 10.1038/322145a0. [DOI] [PubMed] [Google Scholar]
  6. Capone M., Watrin F., Fernex C., Horvat B., Krippl B., Wu L., Scollay R., Ferrier P. TCR beta and TCR alpha gene enhancers confer tissue- and stage-specificity on V(D)J recombination events. EMBO J. 1993 Nov;12(11):4335–4346. doi: 10.1002/j.1460-2075.1993.tb06118.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Castelli C., Mazzocchi A., Salvi S., Anichini A., Sensi M. Use of the V delta 1 variable region in the functional T-cell receptor alpha chain of a WT31+ cytotoxic T lymphocyte clone which specifically recognizes HLA-A2 molecule. Scand J Immunol. 1992 Apr;35(4):487–494. doi: 10.1111/j.1365-3083.1992.tb02883.x. [DOI] [PubMed] [Google Scholar]
  8. Chien Y. H., Iwashima M., Kaplan K. B., Elliott J. F., Davis M. M. A new T-cell receptor gene located within the alpha locus and expressed early in T-cell differentiation. 1987 Jun 25-Jul 1Nature. 327(6124):677–682. doi: 10.1038/327677a0. [DOI] [PubMed] [Google Scholar]
  9. Chien Y. H., Iwashima M., Wettstein D. A., Kaplan K. B., Elliott J. F., Born W., Davis M. M. T-cell receptor delta gene rearrangements in early thymocytes. Nature. 1987 Dec 24;330(6150):722–727. doi: 10.1038/330722a0. [DOI] [PubMed] [Google Scholar]
  10. Dent A. L., Matis L. A., Hooshmand F., Widacki S. M., Bluestone J. A., Hedrick S. M. Self-reactive gamma delta T cells are eliminated in the thymus. Nature. 1990 Feb 22;343(6260):714–719. doi: 10.1038/343714a0. [DOI] [PubMed] [Google Scholar]
  11. Dudley E. C., Girardi M., Owen M. J., Hayday A. C. Alpha beta and gamma delta T cells can share a late common precursor. Curr Biol. 1995 Jun 1;5(6):659–669. doi: 10.1016/s0960-9822(95)00131-x. [DOI] [PubMed] [Google Scholar]
  12. Elliott J. F., Rock E. P., Patten P. A., Davis M. M., Chien Y. H. The adult T-cell receptor delta-chain is diverse and distinct from that of fetal thymocytes. Nature. 1988 Feb 18;331(6157):627–631. doi: 10.1038/331627a0. [DOI] [PubMed] [Google Scholar]
  13. Ferrier P., Krippl B., Blackwell T. K., Furley A. J., Suh H., Winoto A., Cook W. D., Hood L., Costantini F., Alt F. W. Separate elements control DJ and VDJ rearrangement in a transgenic recombination substrate. EMBO J. 1990 Jan;9(1):117–125. doi: 10.1002/j.1460-2075.1990.tb08087.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Garman R. D., Doherty P. J., Raulet D. H. Diversity, rearrangement, and expression of murine T cell gamma genes. Cell. 1986 Jun 6;45(5):733–742. doi: 10.1016/0092-8674(86)90787-7. [DOI] [PubMed] [Google Scholar]
  15. Gascoigne N. R., Chien Y., Becker D. M., Kavaler J., Davis M. M. Genomic organization and sequence of T-cell receptor beta-chain constant- and joining-region genes. Nature. 1984 Aug 2;310(5976):387–391. doi: 10.1038/310387a0. [DOI] [PubMed] [Google Scholar]
  16. Godfrey D. I., Kennedy J., Suda T., Zlotnik A. A developmental pathway involving four phenotypically and functionally distinct subsets of CD3-CD4-CD8- triple-negative adult mouse thymocytes defined by CD44 and CD25 expression. J Immunol. 1993 May 15;150(10):4244–4252. [PubMed] [Google Scholar]
  17. Godfrey D. I., Zlotnik A., Suda T. Phenotypic and functional characterization of c-kit expression during intrathymic T cell development. J Immunol. 1992 Oct 1;149(7):2281–2285. [PubMed] [Google Scholar]
  18. Haskins K., Kubo R., White J., Pigeon M., Kappler J., Marrack P. The major histocompatibility complex-restricted antigen receptor on T cells. I. Isolation with a monoclonal antibody. J Exp Med. 1983 Apr 1;157(4):1149–1169. doi: 10.1084/jem.157.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hata S., Satyanarayana K., Devlin P., Band H., McLean J., Strominger J. L., Brenner M. B., Krangel M. S. Extensive junctional diversity of rearranged human T cell receptor delta genes. Science. 1988 Jun 10;240(4858):1541–1544. doi: 10.1126/science.3259726. [DOI] [PubMed] [Google Scholar]
  20. Hockett R. D., Jr, Nuñez G., Korsmeyer S. J. Evolutionary comparison of murine and human delta T-cell receptor deleting elements. New Biol. 1989 Dec;1(3):266–274. [PubMed] [Google Scholar]
  21. Hockett R. D., de Villartay J. P., Pollock K., Poplack D. G., Cohen D. I., Korsmeyer S. J. Human T-cell antigen receptor (TCR) delta-chain locus and elements responsible for its deletion are within the TCR alpha-chain locus. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9694–9698. doi: 10.1073/pnas.85.24.9694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ishida I., Verbeek S., Bonneville M., Itohara S., Berns A., Tonegawa S. T-cell receptor gamma delta and gamma transgenic mice suggest a role of a gamma gene silencer in the generation of alpha beta T cells. Proc Natl Acad Sci U S A. 1990 Apr;87(8):3067–3071. doi: 10.1073/pnas.87.8.3067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Koop B. F., Rowen L., Wang K., Kuo C. L., Seto D., Lenstra J. A., Howard S., Shan W., Deshpande P., Hood L. The human T-cell receptor TCRAC/TCRDC (C alpha/C delta) region: organization, sequence, and evolution of 97.6 kb of DNA. Genomics. 1994 Feb;19(3):478–493. doi: 10.1006/geno.1994.1097. [DOI] [PubMed] [Google Scholar]
  24. Koop B. F., Wilson R. K., Wang K., Vernooij B., Zallwer D., Kuo C. L., Seto D., Toda M., Hood L. Organization, structure, and function of 95 kb of DNA spanning the murine T-cell receptor C alpha/C delta region. Genomics. 1992 Aug;13(4):1209–1230. doi: 10.1016/0888-7543(92)90039-u. [DOI] [PubMed] [Google Scholar]
  25. Kronenberg M., Siu G., Hood L. E., Shastri N. The molecular genetics of the T-cell antigen receptor and T-cell antigen recognition. Annu Rev Immunol. 1986;4:529–591. doi: 10.1146/annurev.iy.04.040186.002525. [DOI] [PubMed] [Google Scholar]
  26. Lanier L. L., Weiss A. Presence of Ti (WT31) negative T lymphocytes in normal blood and thymus. Nature. 1986 Nov 20;324(6094):268–270. doi: 10.1038/324268a0. [DOI] [PubMed] [Google Scholar]
  27. Lauzurica P., Krangel M. S. Temporal and lineage-specific control of T cell receptor alpha/delta gene rearrangement by T cell receptor alpha and delta enhancers. J Exp Med. 1994 Jun 1;179(6):1913–1921. doi: 10.1084/jem.179.6.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Livak F., Petrie H. T., Crispe I. N., Schatz D. G. In-frame TCR delta gene rearrangements play a critical role in the alpha beta/gamma delta T cell lineage decision. Immunity. 1995 Jun;2(6):617–627. doi: 10.1016/1074-7613(95)90006-3. [DOI] [PubMed] [Google Scholar]
  29. Loh E. Y., Cwirla S., Serafini A. T., Phillips J. H., Lanier L. L. Human T-cell-receptor delta chain: genomic organization, diversity, and expression in populations of cells. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9714–9718. doi: 10.1073/pnas.85.24.9714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. McDonnell T. J., Deane N., Platt F. M., Nunez G., Jaeger U., McKearn J. P., Korsmeyer S. J. bcl-2-immunoglobulin transgenic mice demonstrate extended B cell survival and follicular lymphoproliferation. Cell. 1989 Apr 7;57(1):79–88. doi: 10.1016/0092-8674(89)90174-8. [DOI] [PubMed] [Google Scholar]
  31. Meuer S. C., Fitzgerald K. A., Hussey R. E., Hodgdon J. C., Schlossman S. F., Reinherz E. L. Clonotypic structures involved in antigen-specific human T cell function. Relationship to the T3 molecular complex. J Exp Med. 1983 Feb 1;157(2):705–719. doi: 10.1084/jem.157.2.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ohashi P. S., Wallace V. A., Broughton H., Ohashi C. T., Ferrick D. A., Jost V., Mak T. W., Hengartner H., Pircher H. Specific deletion of the J-C delta locus in murine alpha/beta T cell clones and studies using transgenic mice. Eur J Immunol. 1990 Mar;20(3):517–522. doi: 10.1002/eji.1830200309. [DOI] [PubMed] [Google Scholar]
  33. Pearse M., Wu L., Egerton M., Wilson A., Shortman K., Scollay R. A murine early thymocyte developmental sequence is marked by transient expression of the interleukin 2 receptor. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1614–1618. doi: 10.1073/pnas.86.5.1614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Petrie H. T., Hugo P., Scollay R., Shortman K. Lineage relationships and developmental kinetics of immature thymocytes: CD3, CD4, and CD8 acquisition in vivo and in vitro. J Exp Med. 1990 Dec 1;172(6):1583–1588. doi: 10.1084/jem.172.6.1583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Petrie H. T., Pearse M., Scollay R., Shortman K. Development of immature thymocytes: initiation of CD3, CD4, and CD8 acquisition parallels down-regulation of the interleukin 2 receptor alpha chain. Eur J Immunol. 1990 Dec;20(12):2813–2815. doi: 10.1002/eji.1830201243. [DOI] [PubMed] [Google Scholar]
  36. Raulet D. H., Spencer D. M., Hsiang Y. H., Goldman J. P., Bix M., Liao N. S., Zijstra M., Jaenisch R., Correa I. Control of gamma delta T-cell development. Immunol Rev. 1991 Apr;120:185–204. doi: 10.1111/j.1600-065x.1991.tb00592.x. [DOI] [PubMed] [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Satyanarayana K., Hata S., Devlin P., Roncarolo M. G., De Vries J. E., Spits H., Strominger J. L., Krangel M. S. Genomic organization of the human T-cell antigen-receptor alpha/delta locus. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8166–8170. doi: 10.1073/pnas.85.21.8166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Silverman G. A., Ye R. D., Pollock K. M., Sadler J. E., Korsmeyer S. J. Use of yeast artificial chromosome clones for mapping and walking within human chromosome segment 18q21.3. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7485–7489. doi: 10.1073/pnas.86.19.7485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Suda T., Zlotnik A. Origin, differentiation, and repertoire selection of CD3+CD4-CD8- thymocytes bearing either alpha beta or gamma delta T cell receptors. J Immunol. 1993 Jan 15;150(2):447–455. [PubMed] [Google Scholar]
  41. Takeshita S., Toda M., Yamagishi H. Excision products of the T cell receptor gene support a progressive rearrangement model of the alpha/delta locus. EMBO J. 1989 Nov;8(11):3261–3270. doi: 10.1002/j.1460-2075.1989.tb08486.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Takihara Y., Tkachuk D., Michalopoulos E., Champagne E., Reimann J., Minden M., Mak T. W. Sequence and organization of the diversity, joining, and constant region genes of the human T-cell delta-chain locus. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6097–6101. doi: 10.1073/pnas.85.16.6097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Thompson S. D., Pelkonen J., Hurwitz J. L. Concomitant T-cell receptor alpha and delta gene rearrangements in individual T-cell precursors. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5583–5586. doi: 10.1073/pnas.87.14.5583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Utsunomiya Y., Bill J., Palmer E., Kanagawa O. Identification of a mouse T-cell antigen receptor alpha-chain polymorphism by a V alpha 3.2 chain-specific monoclonal antibody. Immunogenetics. 1991;33(3):198–201. doi: 10.1007/BF01719241. [DOI] [PubMed] [Google Scholar]
  45. Winoto A., Baltimore D. Alpha beta lineage-specific expression of the alpha T cell receptor gene by nearby silencers. Cell. 1989 Nov 17;59(4):649–655. doi: 10.1016/0092-8674(89)90010-x. [DOI] [PubMed] [Google Scholar]
  46. Winoto A., Baltimore D. Separate lineages of T cells expressing the alpha beta and gamma delta receptors. Nature. 1989 Mar 30;338(6214):430–432. doi: 10.1038/338430a0. [DOI] [PubMed] [Google Scholar]
  47. Winoto A., Mjolsness S., Hood L. Genomic organization of the genes encoding mouse T-cell receptor alpha-chain. 1985 Aug 29-Sep 4Nature. 316(6031):832–836. doi: 10.1038/316832a0. [DOI] [PubMed] [Google Scholar]
  48. Wu L., Scollay R., Egerton M., Pearse M., Spangrude G. J., Shortman K. CD4 expressed on earliest T-lineage precursor cells in the adult murine thymus. Nature. 1991 Jan 3;349(6304):71–74. doi: 10.1038/349071a0. [DOI] [PubMed] [Google Scholar]
  49. Zorn A. M., Krieg P. A. PCR analysis of alternative splicing pathways: identification of artifacts generated by heteroduplex formation. Biotechniques. 1991 Aug;11(2):180–184. [PubMed] [Google Scholar]
  50. de Villartay J. P., Hockett R. D., Coran D., Korsmeyer S. J., Cohen D. I. Deletion of the human T-cell receptor delta-gene by a site-specific recombination. Nature. 1988 Sep 8;335(6186):170–174. doi: 10.1038/335170a0. [DOI] [PubMed] [Google Scholar]
  51. de Villartay J. P., Mossalayi D., de Chasseval R., Dalloul A., Debré P. The differentiation of human pro-thymocytes along the TCR-alpha/beta pathway in vitro is accompanied by the site-specific deletion of the TCR-delta locus. Int Immunol. 1991 Dec;3(12):1301–1305. doi: 10.1093/intimm/3.12.1301. [DOI] [PubMed] [Google Scholar]

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