Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Immunobiology

Characterization of the T cell receptor repertoire of neonatal T cells by RT-PCR and single strand conformation polymorphism analysis

Bone Marrow Transplantation volume 26, pages 83–89 (2000)Cite this article

Abstract

We have analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) the individual non-germ line configurations of the T cell receptor (TCR) Vβ chains expressed by T cells from eight individual cord blood specimens. cDNA from each cord blood was amplified using a common primer coupled with a primer specific for each of 22 variable elements of the Vβ chain family and the amplified fragments were separated under high resolution conditions. With cDNA from adult blood (as a control), all of the TCR chains were amplified as a smear consistent with the extensive polyclonality of adult T cells. In contrast, a heterogeneous pattern of amplification was observed with cDNAs from cord blood: only 26.7 ± 21.9% of the 22 Vβ chains analyzed were amplified as a smear. The majority of them were amplified as a discrete number of bands (up to 10) (in 68.2 ± 18.7% of samples) and some of them as a single fragment (4.0 ± 7.8%). Only one of the eight samples analyzed expressed the majority (72.7%) of its Vβ chains as a smear, consistent with an adult-like TCR repertoire. In conclusion, cord blood expressed, on average, a less complex TCR repertoire than adult blood. Bone Marrow Transplantation (2000) 26, 83–89.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2

Similar content being viewed by others

References

  1. Schatz DG, Oettinger MA, Schlissel MS . V(D)J recombination: molecular biology and regulation Ann Rev Immunol 1992 10: 359–383

    Article  CAS  Google Scholar 

  2. LaRocque R, Robinson MA . Diversity in the human T cell receptor beta chain Hum Immunol 1996 48: 3–11

    Article  CAS  Google Scholar 

  3. Wei S, Charmley P, Robinson M, Concannon P . The extent of human germline T-cell receptor Vβ gene segment repertoire Immunogenetics 1994 40: 27–36

    Article  CAS  Google Scholar 

  4. Arden B, Clark SP, Kabelitz D et al. Human T-cell receptor variable gene segment families Immunogenetics 1995 42: 455–500

    CAS  PubMed  Google Scholar 

  5. Alam A, Lulé J, Coppin H et al. T-cell receptor of the β-chain gene use in peripheral blood and multiple synovial membranes during rheumatoid arthritis Hum Immunol 1995 42: 331–339

    Article  CAS  Google Scholar 

  6. Siu G, Clark S, Yoshikai Y et al. The human T-cell antigen receptor is encoded by variable, diversity and joining gene segments that rearrange to generate a complete V gene Cell 1984 37: 393–401

    Article  CAS  Google Scholar 

  7. Gilfillan S, Dierrich A, Lemeur M et al. Mice lacking TdT: mature animals with an immature lymphocyte repertoire Science 1993 261: 1175–1178

    Article  CAS  Google Scholar 

  8. Komori T, Okada A, Stewart V et al. Lack of N regions in antigen receptor variable region genes of TdT-deficient lymphocytes Science 1993 261: 1171–1175

    Article  CAS  Google Scholar 

  9. Meier J, Lewis S . P nucleotides in V(D)J recombination: a fine-structure analysis Mol Cell Biol 1993 13: 1078–1092

    Article  CAS  Google Scholar 

  10. Malissen M, Trucy J, Jouven-Marche E et al. Regulation of TCR α and β genes allelic exclusion during T-cell development Immunol Today 1992 13: 315–322

    Article  CAS  Google Scholar 

  11. Cairo MS, Wagner JE . Placental and/or umbilical cord blood: an alternative source of hematopoietic stem cell for transplantation Blood 1997 90: 4665–4678

    CAS  PubMed  Google Scholar 

  12. Rubinstein P, Rosenfield RE, Adamson JW et al. Stored placental blood for unrelated bone marrow reconstitution Blood 1993 81: 1679–1690

    CAS  PubMed  Google Scholar 

  13. Gluckman E, Rocha V, Boyer-Chammard A et al. Outcome of cord blood transplantation from related and unrelated donors New Engl J Med 1997 337: 373–381

    Article  CAS  Google Scholar 

  14. Rubinstein P, Carrier C, Scaradavou A et al. Outcomes among 562 recipients of placental-blood transplants from unrelated donors New Engl J Med 1998 339: 1565–1577

    Article  CAS  Google Scholar 

  15. Hassan J, Reen DJ . Cord blood CD4+CD45RA+ T cells achieved a lower magnitude of activation when compared with their adult counterparts Immunology 1997 90: 397–401

    Article  CAS  Google Scholar 

  16. Harris DT, Schumacher MJ, Locascio J et al. Phenotypic and functional immaturity of human umbilical cord blood T lymphocytes Proc Natl Acad Sci USA 1992 89: 10006–10010

    Article  CAS  Google Scholar 

  17. Risdon G, Gaddy J, Horie M et al. Alloantigen priming induces a state of unresponsiveness in human umbilical cord blood T cells Proc Natl Acad Sci USA 1995 92: 2413–2417

    Article  CAS  Google Scholar 

  18. Lewis DB, Wilson CB . Developmental immunology and role of host defences in neonatal susceptibility to infection. In: Remington J, Klein J (eds) Infectious Diseases of the Fetus and Newborn Infant Saunders Inc: Toronto 1995 p20

    Google Scholar 

  19. Takahashi N, Imanishi K, Nishida H et al. Evidence for immunologic immaturity of cord blood T cells. Cord blood T cells are susceptible to tolerance induction to in vitro stimulation with a superantigen J Immunol 1995 155: 5213–5219

    CAS  PubMed  Google Scholar 

  20. Clement LT, Yamashita N, Martin AM . The functionally distinct subpopulation of human CD+4 helper/inducer T lymphocytes defined by anti-CD45R antibodies derived sequentially from a differentiation pathway that is regulated by activation dependent post-thymic differentiation J Immunol 1988 141: 1464–1470

    CAS  PubMed  Google Scholar 

  21. Pirenne H, Aujard J, Eljaafari A et al. Comparison of T cell functional changes during childhood with the ontogeny of Cdw29 and CD45RA expression on CD+4 T cells Pediatr Res 1992 31: 81–86

    Article  Google Scholar 

  22. Shiratsuchi H, Tsyuiguchi I . Tubercolin PPD reactive T cells in cord blood lymphocytes Infect Immun 1981 33: 651–657

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Munk ME, Kaufmann SHE . Human cord blood receptor antibody cell response to protein antigens of Paracoccidoides braziliensis yeast forms Immunology 1995 84: 98–104

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Miles EA, Wagner JA, Jones AC et al. Peripheral blood mononuclear cell proliferative responses in the first year of life in babies born to allergic parents Clin Exp Allergy 1996 26: 780–788

    Article  CAS  Google Scholar 

  25. Holt PG . Primary allergic sensitization to environmental antigens: perinatal T cell priming as a determinant of responder phenotype in adulthood J Exp Med 1996 183: 1297–1301

    Article  CAS  Google Scholar 

  26. Trivedi HN, HayGlass KT, Gangur V et al. Analysis of neonatal T cell and antigen presenting cell functions Hum Immunol 1997 57: 69–79

    Article  CAS  Google Scholar 

  27. Campana D, Janossy G, Coustan-Smith E et al. The expression of T cell receptor-associated proteins during T cell ontogenesis in man J Immunol 1989 142: 57–66

    CAS  PubMed  Google Scholar 

  28. Doherty PJ, Roifman CM, Pan S et al. Expression of the human T cell receptor Vβ repertoire Mol Immunol 1991 28: 607–612

    Article  CAS  Google Scholar 

  29. Bonati A, Zanelli P, Ferrari S et al. T-cell receptor β-chain gene rearrangement and expression during human thymic ontogenesis Blood 1992 79: 1472–1483

    CAS  PubMed  Google Scholar 

  30. Schelonka RL, Raaphorst FM, Infante D et al. T cell receptor repertoire diversity and clonal expansion in human neonates Pediatr Res 1998 43: 396–402

    Article  CAS  Google Scholar 

  31. Garderet L, Dulphy N, Douay C et al. The umbilical cord blood αβ T-cell repertoire: characteristics of a polyclonal and naive but completely formed repertoire Blood 1998 91: 340–346

    CAS  PubMed  Google Scholar 

  32. Orita M, Iwahara H, Kanazawa H et al. Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms Proc Natl Acad Sci USA 1989 86: 2766–2770

    Article  CAS  Google Scholar 

  33. Orita M, Suzuki Y, Sekiyama T et al. Rapid and sensitive detection of point mutation and DNA polymorsphisms using the polymerase chain reaction Genomics 1989 5: 874–879

    Article  CAS  Google Scholar 

  34. Shen D-F, Doukhan L, Kalams S et al. High-resolution analysis of T-cell receptor β-chain repertoires using DNA heteroduplex tracking: generally stable, clonal CD8+ expansion in all healthy young adults J Immunol Meth 1998 215: 113–121

    Article  CAS  Google Scholar 

  35. Sanchez M, Alfani E, Visconti G et al. Thymus-independent T-cell differentiation in vitro Br J Haematol 1998 103: 1198–1205

    Article  CAS  Google Scholar 

  36. Choi Y, Kotzin B, Herron L et al. Interaction of Staphylococcus aureus toxin ‘superantigen’ with human T cells Proc Natl Acad Sci USA 1989 86: 8941–8945

    Article  CAS  Google Scholar 

  37. Yamamoto K, Masuko-Hongo K, Tanaka A et al. Establishment and application of a novel T cell clonality analysis using single-strand conformation polymorphim of T cell receptor messenger signals Hum Immunol 1996 48: 23–31

    Article  CAS  Google Scholar 

  38. Maniatis T, Fritsch EF, Sambrook J . Molecular Cloning: A Laboratory Manual, 2nd edn Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY 1989

    Google Scholar 

  39. Yamamoto K, Sakoda H, Nakajima T et al. Accumulation of multiple T cell clonotypes in the synovial lesions of patients with rheumatoid arthritis revealed by a novel clonality analysis Int Immunol 1992 11: 1219–1223

    Article  Google Scholar 

  40. Blumberg RS, Yockey CE, Gross GG et al. Human intestinal intraepithelial lymphocytes are derived from a limited number of T cell clones that utilize multiple Vβ T cell receptor genes J Immunol 1993 150: 5144–5153

    CAS  PubMed  Google Scholar 

  41. Landor M . Maternal-fetal transfer of immunoglobulins Ann Immunol 1995 74: 279–283

    CAS  Google Scholar 

  42. Antin JH, Ferrara JL . Cytokine dysregulation and acute graft-versus-host disease Blood 1992 80: 2964–2968

    CAS  PubMed  Google Scholar 

  43. Madrigal JA, Cohen SBA, Gluckman E et al. Does cord blood transplantation result in lower graft-versus-host disease? It takes more than two to tango Hum Immunol 1997 56: 1–5

    Article  CAS  Google Scholar 

  44. Roncarolo MG, Vaccarino E, Saracco P et al. Immunologic properties of cord blood. In: Broxmeyer HE (ed) Cellular Characteristic of Cord Blood and Cord Blood Transplantation AABB Press: Bethesda, MD 1998 pp67–79

    Google Scholar 

  45. Bofill M, Akbar AN, Salmon M et al. Immature CD45RAlowROlow T cells in the human cord blood J Immunol 1994 152: 5613–5623

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by institutional funds from Istituto Superiore di Sanità, Rome, Italy, and by contract No. BIO4-CT96–0646 of the European Community. Dr E Mannella, director of the Italian Red Cross in Rome, and Dr H Valenzise, Department of Obstetrician and Gynecology, University of Tor Vergata, Rome, are gratefully acknowledged for providing adult peripheral blood and fetal blood, respectively. Prof G D'Agnolo, chairman of the Laboratory of Cell Biology, is gratefully acknowledged for the continuous support of the study.

Author information

Authors and Affiliations

  1. Laboratory of Cell Biology, Istituto Superiore di Sanità, Rome, Italy

    E Alfani, M Sanchez & G Migliaccio

  2. Clinical Biochemistry, Istituto Superiore di Sanità, Rome, Italy

    AR Migliaccio

  3. Ospedale Civile Tivoli, Tivoli, Italy

    AM Passarelli

Authors
  1. E Alfani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. AR Migliaccio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Sanchez
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. AM Passarelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G Migliaccio
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Alfani, E., Migliaccio, A., Sanchez, M. et al. Characterization of the T cell receptor repertoire of neonatal T cells by RT-PCR and single strand conformation polymorphism analysis. Bone Marrow Transplant 26, 83–89 (2000). https://doi.org/10.1038/sj.bmt.1702434

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1702434

Keywords

This article is cited by

Search

Advanced search

Quick links