Skip to main content
NCBI home page
Immunology logoLink to Immunology
. 1993 Feb;78(2):252–259.

Diversity of functional T-cell receptor delta-chain transcripts from bone marrow cells of athymic nude mice.

S Spiess 1, A Kuhröber 1, R Schirmbeck 1, B Arden 1, J Reimann 1
PMCID: PMC1421803  PMID: 8473015

Abstract

CD3+ cells are detectable in bone marrow of athymic mice homozygous for the nude mutation. As previously shown, cells expressing the gamma delta T-cell receptor (TcR) represent 30-40% of this T-cell population. Using V delta-specific, V alpha 4-specific, and C delta-specific primers, TcR delta-chain transcripts were reverse transcribed and polymerase chain reaction (PCR)-amplified from total RNA prepared from bone marrow cells (BMC) of 6-month-old NMRI nu/nu mice. Amplified TcR delta-chain cDNA was cloned, and 49 randomly selected clones derived from seven amplification reactions were sequenced. Sequence analyses showed: (1) more than 80% of the sequenced clones represented in-frame transcripts of the TcR delta-chain; (2) in-frame transcripts containing V delta 1-, V delta 2-, V delta 3-, V delta 4-, V delta 5-, V delta 6- and V alpha 4-gene segments were detectable in nude BMC; (3) V delta 2-, V delta 4- and V delta 5-containing transcripts were more abundant and more diverse than V delta 1- and V delta 3-containing transcripts; (4) extensive N-region diversity was present in the V delta-D delta 2 (N1), D delta 2-D delta 1 (N2) and D delta 1-J delta 1 (N3) junctional regions; (5) P nucleotide additions were present in many transcripts; and (6) unusual truncated, in-frame transcripts with deleted D- and J-region genes were detected. A large potential TcR delta-chain repertoire is thus present in nude BMC.

Full text

PDF
252

Images in this article

254Figure 1
on p.254

Selected References

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

  1. Abo T., Ohteki T., Seki S., Koyamada N., Yoshikai Y., Masuda T., Rikiishi H., Kumagai K. The appearance of T cells bearing self-reactive T cell receptor in the livers of mice injected with bacteria. J Exp Med. 1991 Aug 1;174(2):417–424. doi: 10.1084/jem.174.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abromson-Leeman S. R., Jayaraman S., Dorf M. E. Characterization of T cell clones from an athymic mouse. J Immunol. 1990 Apr 1;144(7):2451–2458. [PubMed] [Google Scholar]
  3. Allison J. P., Havran W. L. The immunobiology of T cells with invariant gamma delta antigen receptors. Annu Rev Immunol. 1991;9:679–705. doi: 10.1146/annurev.iy.09.040191.003335. [DOI] [PubMed] [Google Scholar]
  4. Asarnow D. M., Goodman T., LeFrancois L., Allison J. P. Distinct antigen receptor repertoires of two classes of murine epithelium-associated T cells. Nature. 1989 Sep 7;341(6237):60–62. doi: 10.1038/341060a0. [DOI] [PubMed] [Google Scholar]
  5. Asarnow D. M., Kuziel W. A., Bonyhadi M., Tigelaar R. E., Tucker P. W., Allison J. P. Limited diversity of gamma delta antigen receptor genes of Thy-1+ dendritic epidermal cells. Cell. 1988 Dec 2;55(5):837–847. doi: 10.1016/0092-8674(88)90139-0. [DOI] [PubMed] [Google Scholar]
  6. Bandeira A., Itohara S., Bonneville M., Burlen-Defranoux O., Mota-Santos T., Coutinho A., Tonegawa S. Extrathymic origin of intestinal intraepithelial lymphocytes bearing T-cell antigen receptor gamma delta. Proc Natl Acad Sci U S A. 1991 Jan 1;88(1):43–47. doi: 10.1073/pnas.88.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Benveniste P., Chadwick B. S., Miller R. G., Reimann J. Characterization of cells with T cell markers in athymic nude bone marrow and of their in vitro-derived clonal progeny. Comparison with euthymic bone marrow. J Immunol. 1990 Jan 15;144(2):411–419. [PubMed] [Google Scholar]
  8. Bonneville M., Itohara S., Krecko E. G., Mombaerts P., Ishida I., Katsuki M., Berns A., Farr A. G., Janeway C. A., Jr, Tonegawa S. Transgenic mice demonstrate that epithelial homing of gamma/delta T cells is determined by cell lineages independent of T cell receptor specificity. J Exp Med. 1990 Apr 1;171(4):1015–1026. doi: 10.1084/jem.171.4.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bosma M. J., Carroll A. M. The SCID mouse mutant: definition, characterization, and potential uses. Annu Rev Immunol. 1991;9:323–350. doi: 10.1146/annurev.iy.09.040191.001543. [DOI] [PubMed] [Google Scholar]
  10. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  11. Brenner M. B., Strominger J. L., Krangel M. S. The gamma delta T cell receptor. Adv Immunol. 1988;43:133–192. [PubMed] [Google Scholar]
  12. Chen W. F., Scollay R., Shortman K., Skinner M., Marbrook J. T-cell development in the absence of a thymus: the number, the phenotype, and the functional capacity of T lymphocytes in nude mice. Am J Anat. 1984 Jul;170(3):339–347. doi: 10.1002/aja.1001700309. [DOI] [PubMed] [Google Scholar]
  13. De Geus B., Van den Enden M., Coolen C., Nagelkerken L., Van der Heijden P., Rozing J. Phenotype of intraepithelial lymphocytes in euthymic and athymic mice: implications for differentiation of cells bearing a CD3-associated gamma delta T cell receptor. Eur J Immunol. 1990 Feb;20(2):291–298. doi: 10.1002/eji.1830200210. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Ezquerra A., Cron R. Q., McConnell T. J., Valas R. B., Bluestone J. A., Coligan J. E. T cell receptor delta-gene expression and diversity in the mouse spleen. J Immunol. 1990 Sep 1;145(5):1311–1317. [PubMed] [Google Scholar]
  16. Feeney A. J. Junctional sequences of fetal T cell receptor beta chains have few N regions. J Exp Med. 1991 Jul 1;174(1):115–124. doi: 10.1084/jem.174.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fujihashi K., Kiyono H., Aicher W. K., Green D. R., Singh B., Eldridge J. H., McGhee J. R. Immunoregulatory function of CD3+, CD4-, and CD8- T cells. Gamma delta T cell receptor-positive T cells from nude mice abrogate oral tolerance. J Immunol. 1989 Dec 1;143(11):3415–3422. [PubMed] [Google Scholar]
  18. Gillis S., Union N. A., Baker P. E., Smith K. A. The in vitro generation and sustained culture of nude mouse cytolytic T-lymphocytes. J Exp Med. 1979 Jun 1;149(6):1460–1476. doi: 10.1084/jem.149.6.1460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Guy-Grand D., Cerf-Bensussan N., Malissen B., Malassis-Seris M., Briottet C., Vassalli P. Two gut intraepithelial CD8+ lymphocyte populations with different T cell receptors: a role for the gut epithelium in T cell differentiation. J Exp Med. 1991 Feb 1;173(2):471–481. doi: 10.1084/jem.173.2.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Guy-Grand D., Malassis-Seris M., Briottet C., Vassalli P. Cytotoxic differentiation of mouse gut thymodependent and independent intraepithelial T lymphocytes is induced locally. Correlation between functional assays, presence of perforin and granzyme transcripts, and cytoplasmic granules. J Exp Med. 1991 Jun 1;173(6):1549–1552. doi: 10.1084/jem.173.6.1549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Haskins K., Kappler J., Marrack P. The major histocompatibility complex-restricted antigen receptor on T cells. Annu Rev Immunol. 1984;2:51–66. doi: 10.1146/annurev.iy.02.040184.000411. [DOI] [PubMed] [Google Scholar]
  22. Havran W. L., Allison J. P. Origin of Thy-1+ dendritic epidermal cells of adult mice from fetal thymic precursors. Nature. 1990 Mar 1;344(6261):68–70. doi: 10.1038/344068a0. [DOI] [PubMed] [Google Scholar]
  23. Hodes R. J., Sharrow S. O., Solomon A. Failure of T cell receptor V beta negative selection in an athymic environment. Science. 1989 Nov 24;246(4933):1041–1044. doi: 10.1126/science.2587987. [DOI] [PubMed] [Google Scholar]
  24. Hurwitz J. L., McAndrew E. C., Doherty P. C. Anti-Thy-1 plus complement-treated, cultured bone marrow cells resemble fetal thymocytes in killer cell function and marker expression. J Immunol. 1986 Sep 15;137(6):1757–1763. [PubMed] [Google Scholar]
  25. Hurwitz J. L., Samaridis J., Pelkonen J. Progression of rearrangements at T cell receptor beta and gamma gene loci during athymic differentiation of bone marrow cells in vitro. Cell. 1988 Mar 25;52(6):821–829. doi: 10.1016/0092-8674(88)90424-2. [DOI] [PubMed] [Google Scholar]
  26. Hünig T., Bevan M. J. Specificity of cytotoxic T cells from athymic mice. J Exp Med. 1980 Sep 1;152(3):688–702. doi: 10.1084/jem.152.3.688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kienker L. J., Kuziel W. A., Tucker P. W. T cell receptor gamma and delta gene junctional sequences in SCID mice: excessive P nucleotide insertion. J Exp Med. 1991 Oct 1;174(4):769–773. doi: 10.1084/jem.174.4.769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kishihara K., Yoshikai Y., Matsuzaki G., Mak T. W., Nomoto K. Functional alpha and beta T cell chain receptor messages can be detected in old but not in young athymic mice. Eur J Immunol. 1987 Apr;17(4):477–482. doi: 10.1002/eji.1830170407. [DOI] [PubMed] [Google Scholar]
  29. Kung J. T. Impaired clonal expansion in athymic nude CD8+CD4- T cells. J Immunol. 1988 Jun 1;140(11):3727–3735. [PubMed] [Google Scholar]
  30. Kung J. T., Thomas C. A., 3rd Athymic nude CD4+8- T cells produce IL-2 but fail to proliferate in response to mitogenic stimuli. J Immunol. 1988 Dec 1;141(11):3691–3696. [PubMed] [Google Scholar]
  31. Lafaille J. J., DeCloux A., Bonneville M., Takagaki Y., Tonegawa S. Junctional sequences of T cell receptor gamma delta genes: implications for gamma delta T cell lineages and for a novel intermediate of V-(D)-J joining. Cell. 1989 Dec 1;59(5):859–870. doi: 10.1016/0092-8674(89)90609-0. [DOI] [PubMed] [Google Scholar]
  32. Lake J. P., Pierce C. W., Kennedy J. D. T cell receptor expression by T cells that mature extrathymically in nude mice. Cell Immunol. 1991 Jun;135(1):259–265. doi: 10.1016/0008-8749(91)90270-l. [DOI] [PubMed] [Google Scholar]
  33. Lawetzky A., Hünig T. Analysis of CD3 and antigen receptor expression on T cell subpopulations of aged athymic mice. Eur J Immunol. 1988 Mar;18(3):409–416. doi: 10.1002/eji.1830180314. [DOI] [PubMed] [Google Scholar]
  34. MacDonald H. R., Blanc C., Lees R. K., Sordat B. Abnormal distribution of T cell subsets in athymic mice. J Immunol. 1986 Jun 15;136(12):4337–4339. [PubMed] [Google Scholar]
  35. MacDonald H. R., Lees R. K., Bron C., Sordat B., Miescher G. T cell antigen receptor expression in athymic (nu/nu) mice. Evidence for an oligoclonal beta chain repertoire. J Exp Med. 1987 Jul 1;166(1):195–209. doi: 10.1084/jem.166.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. MacDonald H. R., Lees R. K., Sordat B., Zaech P., Maryanski J. L., Bron C. Age-associated increase in expression of the T cell surface markers Thy-1, Lyt-1, and Lyt-2 in congenitally athymic (nu/nu) mice: analysis by flow microfluorometry. J Immunol. 1981 Mar;126(3):865–870. [PubMed] [Google Scholar]
  37. Maleckar J. R., Sherman L. A. The composition of the T cell receptor repertoire in nude mice. J Immunol. 1987 Jun 1;138(11):3873–3876. [PubMed] [Google Scholar]
  38. Maryanski J. L., MacDonald H. R., Sordat B., Cerottini J. C. Cytolytic T lymphocyte precursor cells in congenitally athymic C57BL/6 nu/nu mice: quantitation, enrichment, and specificity. J Immunol. 1981 Mar;126(3):871–876. [PubMed] [Google Scholar]
  39. Miller R. G., Derry H. R., Sarjeant B. J. The extent of self-MHC restriction of cytotoxic T cells in nude mice varies from mouse to mouse. J Immunol. 1983 Jan;130(1):63–66. [PubMed] [Google Scholar]
  40. Mosley R. L., Styre D., Klein J. R. Differentiation and functional maturation of bone marrow-derived intestinal epithelial T cells expressing membrane T cell receptor in athymic radiation chimeras. J Immunol. 1990 Sep 1;145(5):1369–1375. [PubMed] [Google Scholar]
  41. Nandi D., Allison J. P. Phenotypic analysis and gamma delta-T cell receptor repertoire of murine T cells associated with the vaginal epithelium. J Immunol. 1991 Sep 15;147(6):1773–1778. [PubMed] [Google Scholar]
  42. Ohteki T., Seki S., Abo T., Kumagai K. Liver is a possible site for the proliferation of abnormal CD3+4-8- double-negative lymphocytes in autoimmune MRL-lpr/lpr mice. J Exp Med. 1990 Jul 1;172(1):7–12. doi: 10.1084/jem.172.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Reimann J., Miller R. G. Differentiation from precursors in athymic nude mouse bone marrow of unusual spontaneously cytolytic cells showing anti-self-H-2 specificity and bearing T cell markers. J Exp Med. 1983 Nov 1;158(5):1672–1692. doi: 10.1084/jem.158.5.1672. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rocha B. Characterization of V beta-bearing cells in athymic (nu/nu) mice suggests an extrathymic pathway for T cell differentiation. Eur J Immunol. 1990 Apr;20(4):919–925. doi: 10.1002/eji.1830200430. [DOI] [PubMed] [Google Scholar]
  45. Rocha B., Vassalli P., Guy-Grand D. The V beta repertoire of mouse gut homodimeric alpha CD8+ intraepithelial T cell receptor alpha/beta + lymphocytes reveals a major extrathymic pathway of T cell differentiation. J Exp Med. 1991 Feb 1;173(2):483–486. doi: 10.1084/jem.173.2.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Schuler W., Weiler I. J., Schuler A., Phillips R. A., Rosenberg N., Mak T. W., Kearney J. F., Perry R. P., Bosma M. J. Rearrangement of antigen receptor genes is defective in mice with severe combined immune deficiency. Cell. 1986 Sep 26;46(7):963–972. doi: 10.1016/0092-8674(86)90695-1. [DOI] [PubMed] [Google Scholar]
  47. Seki S., Abo T., Masuda T., Ohteki T., Kanno A., Takeda K., Rikiishi H., Nagura H., Kumagai K. Identification of activated T cell receptor gamma delta lymphocytes in the liver of tumor-bearing hosts. J Clin Invest. 1990 Aug;86(2):409–415. doi: 10.1172/JCI114726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Sim G. K., Augustin A. Dominantly inherited expression of BID, an invariant undiversified T cell receptor delta chain. Cell. 1990 May 4;61(3):397–405. doi: 10.1016/0092-8674(90)90522-g. [DOI] [PubMed] [Google Scholar]
  49. Sydora B. C., Kronenberg M. Characterization of a CD4-positive T-cell line derived from an athymic (nu/nu) mouse. Cell Immunol. 1991 Apr 15;134(1):54–64. doi: 10.1016/0008-8749(91)90330-e. [DOI] [PubMed] [Google Scholar]
  50. Takagaki Y., DeCloux A., Bonneville M., Tonegawa S. Diversity of gamma delta T-cell receptors on murine intestinal intra-epithelial lymphocytes. Nature. 1989 Jun 29;339(6227):712–714. doi: 10.1038/339712a0. [DOI] [PubMed] [Google Scholar]
  51. Yoshikai Y., Matsuzaki G., Takeda Y., Ohga S., Kishihara K., Yuuki H., Nomoto K. Functional T cell receptor delta chain gene messages in athymic nude mice. Eur J Immunol. 1988 Jul;18(7):1039–1043. doi: 10.1002/eji.1830180711. [DOI] [PubMed] [Google Scholar]
  52. Yoshikai Y., Reis M. D., Mak T. W. Athymic mice express a high level of functional gamma-chain but greatly reduced levels of alpha- and beta-chain T-cell receptor messages. Nature. 1986 Dec 4;324(6096):482–485. doi: 10.1038/324482a0. [DOI] [PubMed] [Google Scholar]
  53. 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 Immunology are provided here courtesy of British Society for Immunology

RESOURCES