Immunologic Research

, Volume 23, Issue 1, pp 75–84 | Cite as

Individuality of Ag-selected and preimmune TCR repertoires

  • Janet L. Maryanski
  • Valérie Attuil
  • Abdelbasset Hamrouni
  • Mireille Mutin
  • Mauricette Rossi
  • Anne Aublin
  • Philipp Bucher


We have combined flow cytometry and single-cell PCR to characterize the TCR BV repertoires selected by individual mice in a model CD8 response against a defined peptide/MHC complex (CW 3 170–179/Kd). Ourresults established that different mice select individually distinct yet structurally similar CW3-specific repertoires. Repertoire selection appears to be flexible depending on the immunizing cell dose. Using a single-donor, matched-pair-recipient adoptive transfer strategy, we demonstrated that the CW3-specific TCR repertoires of normal mice are already distinct at the preimmune level. We combine our data with computer simulations to test models for the composition of an Ag-specific preimmune repertoire and its selection during an immune response.

Key Words

TCR sequences Ag-selected repertoire Preimmune repertoire Single-cell PCR Adoptive transfer Computer simulations 


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  1. 1.
    Davis MM, Bjorkman PJ: T-cell antigen receptor genes and T-cell recognition. Nature 1988;334:395–402.PubMedCrossRefGoogle Scholar
  2. 2.
    Garcia KC, Teyton L, Wilson IA: Structural basis of T cell recognition. Annu Rev Immunol 1999; 17:369–397.PubMedCrossRefGoogle Scholar
  3. 3.
    Rotzschke O, Falk K, Deres K, Schild H, Norda M, Metzger J, Jung G, Rammensee HG: Isolation and analysis of naturally processed viral peptides as recognized by cytotoxic T cells. Nature 1990; 348:252–254.PubMedCrossRefGoogle Scholar
  4. 4.
    Townsend AR, Rothbard J, Gotch FM, Bahadur G, Wraith D, McMichael AJ: The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell 1986;44;959–968.PubMedCrossRefGoogle Scholar
  5. 5.
    Amsen D, Kruisbeek AM: Thymocyte selection: not by TCR alone. Immunol Rev 1998;165: 209–229PubMedCrossRefGoogle Scholar
  6. 6.
    Davis MM: T cell receptor gene diversity and selection. Annu Rev Biochem 1990;59:475–496.PubMedCrossRefGoogle Scholar
  7. 7.
    Sebzda E, Mariathasan S, Ohteki T, Jones R, Bachmann MF, Ohashi PS: Selection of the T cell repertoire. Annu. Rev. Immunol. 1999; 17:829–874.PubMedCrossRefGoogle Scholar
  8. 8.
    Casrouge A, Beaudoing E, Dalle S, Pannetier C, Kanellopoulos J, Kourilsky P: Size estimate of the alpha beta TCR repertoire of naive mouse splenocytes. J Immunol 2000;164:5782–5787.PubMedGoogle Scholar
  9. 9.
    Arstila TP, Casrouge A, Baron V, Even J, Kanellopoulos J, Kourilsky P: A direct estimate of the human alphabeta T cell receptor diversity. Science 1999;286:958–961.PubMedCrossRefGoogle Scholar
  10. 10.
    Casanova JL, Maryanski JL: Antigen-selected T-cell receptor diversity and self-nonself homology. Immunol Today 1993;14:391–394.PubMedCrossRefGoogle Scholar
  11. 11.
    Kranz DM, Tjoa B: αβ T-cell receptor repertoires among cytotoxic and helper T lymphocytes: in M. Sitkovsky and P. Henkart (eds): Cytotoxic Cells. Boston, Birkhäuser, 1993, 49–57.Google Scholar
  12. 12.
    Casanova JL, Romero P, Widmann C, Kourilsky P, Maryanski JL: T cell receptor genes in a series of class 1 major histocompatibility complex-restricted cytotoxic T lymphocyte clones speqcific for a Plasmodium berghe inonapeptide: Implications for T cell allelic exclusion and antigen-specific repertoire. J Exp Med 1991;174: 1371–1383PubMedCrossRefGoogle Scholar
  13. 13.
    Boitel B, Ermonval M, Panina-Bordigmon P, Mariuzza RA, Lanzavecchia A, Acuto O: Preferential V beta gene usage and lack of junctional sequence conservation among human T cell receptors specific for a tetanus toxin-derived peptide: evidence for a dominant role of a germline-encoded V region in antigen/major histocompatibility complex recognition. J Exp Med 1992;175:765–777.PubMedCrossRefGoogle Scholar
  14. 14.
    Kurokawa M, Furukawa H, Yabe T, Matsui T, Toda M, Hamada C, Kasukawa R, Yamamoto K, Nishioka K, Kato T: Frequency of clonally expanded T cells evaluated by PCR from a single cell. J Immunol Methods 1999;224:203–208.PubMedCrossRefGoogle Scholar
  15. 15.
    Altman JD, Moss P, Goulder P jr., Barouch DH, McHeyzer-Williams MG, Bell JL, McMichael AIJ, Davis MM: Phenotypic analysis of antigen-specific T lymphocytes. Science 1996;274:94–96.PubMedCrossRefGoogle Scholar
  16. 16.
    Casanova JL, Cerottini JC, Matthes M, Necker A, Gournier H, Barra C, Widmann C, MacDonald HR, Lemonnier F, Malissen B, et al.: H-2-restricted cytolytic T lymphocytes specific for HLA display T cell receptors of limited diversity. J Exp Med 1992;176:439–447.PubMedCrossRefGoogle Scholar
  17. 17.
    MacDonald HR, Casanova JL, Maryanski JL, Cerottini JC: Oligoclonal expansion of major histocompatibility complex class 1-restricted cytolytic T lymphocytes during a primary immune response in vivo: direct monitoring by flow cytometry and polymerase chain reaction. J Exp Med 1993;177:1487–1492PubMedCrossRefGoogle Scholar
  18. 18.
    Walker PR, Ohteki T, Lopez JA, MacDonald HR, Maryanski JL: Distinct phenotypes of antigen-selected CD*T cells emerge at different stages of an in vivo immune response. J Immunol 1995;155:3443–3452.PubMedGoogle Scholar
  19. 19.
    Maryanski JL, Jongeneel CV, Bucher P, Casanova JL, Walker PR: Single-cell PCR analysis of TCR repertoires selected by antigen in vivo: a high magnitude CD8 response is comprised of very few clones. Immunity 1996;4:47–55.PubMedCrossRefGoogle Scholar
  20. 20.
    McHeyzer-Williams MG, Davis MM: Antigen-specific development of primary and memory T cells in vivo. Science 1995;268:106–111.PubMedCrossRefGoogle Scholar
  21. 21.
    Maryanski JL, Accolla RS, Jordan B: H2-restricted recognition of cloned HLA class I gene products expressed in mouse cells. J Immunol 1986;136:4340–4347.PubMedGoogle Scholar
  22. 22.
    Maryanski JL, Pala P, Corradin G, Jordan BR, Cerottini JC: H-2-restricted cytolytic T cells specific for HLA can recognize a synthetic HLA peptide. Nature 1986;324:578–579.PubMedCrossRefGoogle Scholar
  23. 23.
    Maryanski JL, Casanova JL, Falk K, Gournier H, Jaulin C, Kourilsky P, Lemonnier FA, Luthy R, Rammensee HG, Rotzschke O, Servis C, Lopez JA: The diversity of antigen-specific TCR repertoires reflects the relative complexity of epitopes recognized. Hum Immunol 1997;54:117–128.PubMedCrossRefGoogle Scholar
  24. 24.
    Casanova JL, Martinon F, Gournier H, Barra C, Pannetier C, Regnault A, Kourilsky P, Cerottini JC, Maryanski JL: T cell receptor selection by and recognition of two class I major histocompatibility complex-restricted antigenic peptides that differ at a single position. J Exp Med 1993;177:811–820.PubMedCrossRefGoogle Scholar
  25. 25.
    Maryanski JL, Attuil V, Bucher P, Walker PR: A quantitative, single-cell PCR analysis of an antigen-specific TCR repertoire selected during an in vivo CD8 response: directevidence for a wide range of clonesizes with uniform tissue distribution. Mol Immunol 1999;36:745–753.PubMedCrossRefGoogle Scholar
  26. 26.
    Attuil V, Bucher P, Rossi M, Mutin M, Maryanski JL: Comparative T cell receptor repertoire selection by antigen after adoptive transfer: a glimpse at an antigen-specific preimmune repertoire. Proc Natl Acad Sci USA 2000;97:8473–8478.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Janet L. Maryanski
    • 2
  • Valérie Attuil
    • 2
  • Abdelbasset Hamrouni
    • 2
  • Mireille Mutin
    • 2
  • Mauricette Rossi
    • 2
  • Anne Aublin
    • 2
  • Philipp Bucher
    • 1
  1. 1.Experimental Cancer ResearchSwiss Institute of Bioinformatics and Swiss Institute forEpalingesSwitzerland
  2. 2.Iserm Unit 503Lyon Cedex 07France

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