Article | Published:

Human γδ T cells are quickly reconstituted after stem-cell transplantation and show adaptive clonal expansion in response to viral infection

Nature Immunology volume 18, pages 393401 (2017) | Download Citation

This article has been updated

Abstract

To investigate how the human γδ T cell pool is shaped during ontogeny and how it is regenerated after transplantation of hematopoietic stem cells (HSCs), we applied an RNA-based next-generation sequencing approach to monitor the dynamics of the repertoires of γδ T cell antigen receptors (TCRs) before and after transplantation in a prospective cohort study. We found that repertoires of rearranged genes encoding γδ TCRs (TRG and TRD) in the peripheral blood of healthy adults were stable over time. Although a large fraction of human TRG repertoires consisted of public sequences, the TRD repertoires were private. In patients undergoing HSC transplantation, γδ T cells were quickly reconstituted; however, they had profoundly altered TCR repertoires. Notably, the clonal proliferation of individual virus-reactive γδ TCR sequences in patients with reactivation of cytomegalovirus revealed strong evidence for adaptive anti-viral γδ T cell immune responses.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Change history

  • 14 February 2018

    In the version of this Article originally published, in Acknowledgments section "Deutsche José Carreras Leukämie-Stiftung e.V. (DJCLS R12/29 to C.K. and I.P.)", text were missing. The text has been included in PDF and XML. These have been corrected after print.

Accessions

Primary accessions

Sequence Read Archive

References

  1. 1.

    , , , & Peripheral selection of Vδ1+ cells with restricted T cell receptor δ gene junctional repertoire in the peripheral blood of healthy donors. J. Exp. Med. 178, 121–127 (1993).

  2. 2.

    , , , & The δ T cell receptor repertoire in human colon and peripheral blood is oligoclonal irrespective of V region usage. J. Clin. Invest. 96, 1108–1117 (1995).

  3. 3.

    , , , & The Vδ1 T cell receptor repertoire in human small intestine and colon. J. Exp. Med. 180, 183–190 (1994).

  4. 4.

    , & Autoreactivity by design: innate B and T lymphocytes. Nat. Rev. Immunol. 1, 177–186 (2001).

  5. 5.

    & Ontogeny of innate T lymphocytes - some innate lymphocytes are more innate than others. Front. Immunol. 5, 486 (2014).

  6. 6.

    et al. γδ T cells exhibit multifunctional and protective memory in intestinal tissues. Immunity 39, 184–195 (2013).

  7. 7.

    et al. Staphylococcus aureus infection of mice expands a population of memory γδ T cells that are protective against subsequent infection. J. Immunol. 192, 3697–3708 (2014).

  8. 8.

    , & Inflammation induces dermal Vγ4+ γδ T17 memory-like cells that travel to distant skin and accelerate secondary IL-17-driven responses. Proc. Natl. Acad. Sci. USA 112, 8046–8051 (2015).

  9. 9.

    , , , & Dermal IL-17-producing γδ T cells establish long-lived memory in the skin. Eur. J. Immunol. 45, 3022–3033 (2015).

  10. 10.

    et al. Deep sequencing of the human TCRγ and TCRβ repertoires suggests that TCRβ rearranges after αβ and γδ T cell commitment. Sci. Transl. Med. 3, 90ra61 (2011).

  11. 11.

    , , , & A comparison of deep sequencing of TCRG rearrangements vs traditional capillary electrophoresis for assessment of clonality in T-cell lymphoproliferative disorders. Am. J. Clin. Pathol. 141, 348–359 (2014).

  12. 12.

    , , , & Deep sequencing of the T-cell receptor repertoire demonstrates polyclonal T-cell infiltrates in psoriasis. F1000 Res. 4, 460 (2015).

  13. 13.

    , , & Molecular analysis of human γ/δ+ clones from thymus and peripheral blood. J. Exp. Med. 170, 1521–1535 (1989).

  14. 14.

    , , & Regulated expression and structure of T cell receptor γ/δ transcripts in human thymic ontogeny. EMBO J. 10, 83–91 (1991).

  15. 15.

    et al. Peripheral selection of antigen receptor junctional features in a major human γδ subset. Eur. J. Immunol. 23, 804–808 (1993).

  16. 16.

    , , , & Reconstitution of γδ T cell repertoire diversity after human allogeneic hematopoietic cell transplantation and the role of peripheral expansion of mature T cell population in the graft. Bone Marrow Transplant. 26, 177–185 (2000).

  17. 17.

    et al. Skewed T cell receptor repertoire of Vδ1+ γδ T lymphocytes after human allogeneic haematopoietic stem cell transplantation and the potential role for Epstein-Barr virus-infected B cells in clonal restriction. Clin. Exp. Immunol. 149, 70–79 (2007).

  18. 18.

    et al. Development of interleukin-17-producing γδ T cells is restricted to a functional embryonic wave. Immunity 37, 48–59 (2012).

  19. 19.

    et al. Long term disease-free survival in acute leukemia patients recovering with increased γδ T cells after partially mismatched related donor bone marrow transplantation. Bone Marrow Transplant. 39, 751–757 (2007).

  20. 20.

    et al. γδ T cells elicited by CMV reactivation after allo-SCT cross-recognize CMV and leukemia. Leukemia 27, 1328–1338 (2013).

  21. 21.

    , & Cancer immunotherapy using γδ T cells: dealing with diversity. Front. Immunol. 5, 601 (2014).

  22. 22.

    et al. γδ T-cell reconstitution after HLA-haploidentical hematopoietic transplantation depleted of TCR-αβ+/CD19+ lymphocytes. Blood 125, 2349–2358 (2015).

  23. 23.

    , , , & Hunting for clinical translation with innate-like immune cells and their receptors. Leukemia 28, 1181–1190 (2014).

  24. 24.

    et al. Implication of γδ T cells in the human immune response to cytomegalovirus. J. Clin. Invest. 103, 1437–1449 (1999).

  25. 25.

    et al. Major expansion of γδ T lymphocytes following cytomegalovirus infection in kidney allograft recipients. J. Infect. Dis. 179, 1–8 (1999).

  26. 26.

    et al. The role of Vδ2-negative γδ T cells during cytomegalovirus reactivation in recipients of allogeneic stem cell transplantation. Blood 116, 2164–2172 (2010).

  27. 27.

    & γδ T cell receptors. Cell. Mol. Life Sci. 63, 2089–2094 (2006).

  28. 28.

    Human γδ T cells: From a neglected lymphocyte population to cellular immunotherapy: A personal reflection of 30years of γδ T cell research. Clin. Immunol. 172, 90–97 (2016).

  29. 29.

    et al. Effector Vγ9Vδ2 T cells dominate the human fetal γδ T-cell repertoire. Proc. Natl. Acad. Sci. USA 112, E556–E565 (2015).

  30. 30.

    et al. Impact of age, gender, and race on circulating γδ T cells. Hum. Immunol. 71, 968–975 (2010).

  31. 31.

    et al. Selective outgrowth of CD45RO+ Vγ9+/Vδ2+ T-cell receptor γ/δ T cells early after bone marrow transplantation. Blood 78, 1875–1881 (1991).

  32. 32.

    et al. Alteration of the T cell repertoire after bone marrow transplantation. Bone Marrow Transplant. 13, 19–26 (1994).

  33. 33.

    et al. A clonotypic Vγ4Jγ1/Vδ5Dδ2Jδ1 innate γδ T-cell population restricted to the CCR6+CD27 subset. Nat. Commun. 6, 6477 (2015).

  34. 34.

    , & Vγ2Vδ2 T cell receptor recognition of prenyl pyrophosphates is dependent on all CDRs. J. Immunol. 184, 6209–6222 (2010).

  35. 35.

    & Evolution and function of the TCR Vγ9 chain repertoire: It's good to be public. Cell. Immunol. 296, 22–30 (2015).

  36. 36.

    et al. Evidence for extrathymic changes in the T cell receptor γ/δ repertoire. J. Exp. Med. 171, 1597–1612 (1990).

  37. 37.

    , , , & Sex-specific phenotypical and functional differences in peripheral human Vγ9/Vδ2 T cells. J. Leukoc. Biol. 79, 663–666 (2006).

  38. 38.

    et al. In vitro stimulation with a non-peptidic alkylphosphate expands cells expressing Vγ2-Jγ1.2/Vδ2 T-cell receptors. Immunology 104, 19–27 (2001).

  39. 39.

    et al. High frequency of circulating γ δ T cells with dominance of the vδ1 subset in a healthy population. Int. Immunol. 12, 797–805 (2000).

  40. 40.

    et al. T cell receptor γδ repertoire in HIV-1-infected individuals. Eur. J. Immunol. 24, 3044–3049 (1994).

  41. 41.

    , , & The γδ T-cell receptor repertoire is reconstituted in HIV patients after prolonged antiretroviral therapy. AIDS 27, 1557–1562 (2013).

  42. 42.

    et al. Improved immune recovery after transplantation of TCRαβ/CD19-depleted allografts from haploidentical donors in pediatric patients. Bone Marrow Transplant. 50, S6–S10 (2015).

  43. 43.

    & Molecular and cellular insights into T cell exhaustion. Nat. Rev. Immunol. 15, 486–499 (2015).

  44. 44.

    et al. Shared reactivity of Vδ2neg γδ T cells against cytomegalovirus-infected cells and tumor intestinal epithelial cells. J. Exp. Med. 201, 1567–1578 (2005).

  45. 45.

    et al. CMV-independent lysis of glioblastoma by ex vivo expanded/activated Vδ1+ γδ T cells. PLoS One 8, e68729 (2013).

  46. 46.

    et al. Donor Vδ1+ γδ T cells expand after allogeneic hematopoietic stem cell transplantation and show reactivity against CMV-infected cells but not against progressing B-CLL. Exp. Hematol. Oncol. 2, 14 (2013).

  47. 47.

    et al. Cytomegalovirus and tumor stress surveillance by binding of a human γδ T cell antigen receptor to endothelial protein C receptor. Nat. Immunol. 13, 872–879 (2012).

  48. 48.

    et al. Human cytomegalovirus elicits fetal γδ T cell responses in utero. J. Exp. Med. 207, 807–821 (2010).

  49. 49.

    et al. Control of murine cytomegalovirus infection by γδ T cells. PLoS Pathog. 11, e1004481 (2015).

  50. 50.

    γδ T cells confer protection against murine cytomegalovirus (MCMV). PLoS Pathog. 11, e1004702 (2015).

  51. 51.

    , , & IMGT((R)) tools for the nucleotide analysis of immunoglobulin (IG) and T cell receptor (TR) V-(D)-J repertoires, polymorphisms, and IG mutations: IMGT/V-QUEST and IMGT/HighV-QUEST for NGS. Methods Mol. Biol. 882, 569–604 (2012).

  52. 52.

    et al. tcR: an R package for T cell receptor repertoire advanced data analysis. BMC Bioinformatics 16, 175 (2015).

Download references

Acknowledgements

We thank E. Hage and T. Schulz for assistance from the central project Z1, an NGS core facility of Collaborative Research Centre SFB900; C. Struckmann and M. Ballmaier for technical guidance and single-cell sorting; the Hannover Unified Biobank of Hannover Medical School; A. Krueger for reading and criticizing the manuscript; and J. Blume for help in cord-blood preparation. Supported by Deutsche Forschungsgemeinschaft, (SFB900/B8 to C.K. and I.P.), Deutsche José Carreras Leukämie-Stiftung e.V. (DJCLS R12/29 to C.K. and I.P.) and PR727/4-1 to I.P.) and the German Federal Ministry of Education and Research (01EO1302 to C.S.-F., C.K. and I.P.).

Author information

Author notes

    • Christian Koenecke
    •  & Immo Prinz

    These authors contributed equally to this work.

Affiliations

  1. Institute of Immunology, Hannover Medical School, Hannover, Germany.

    • Sarina Ravens
    • , Christian Schultze-Florey
    • , Solaiman Raha
    • , Inga Sandrock
    • , Linda Oberdörfer
    • , Annika Reinhardt
    • , Inga Ravens
    • , Maleen Beck
    • , Reinhold Förster
    • , Christian Koenecke
    •  & Immo Prinz
  2. Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.

    • Christian Schultze-Florey
    • , Melanie Drenker
    • , Maleen Beck
    • , Michael Heuser
    • , Felicitas Thol
    • , Arnold Ganser
    •  & Christian Koenecke
  3. Integrated Research and Treatment Center Transplantation, Hannover Medical School, Hannover, Germany.

    • Christian Schultze-Florey
    • , Christian Koenecke
    •  & Immo Prinz
  4. Genome Analytics, Helmholtz Centre for Infection Research, Braunschweig, Germany.

    • Robert Geffers
  5. Department Obstetrics, Gynecology and Reproductive Medicine, Hannover Medical School, Hannover, Germany.

    • Constantin von Kaisenberg

Authors

  1. Search for Sarina Ravens in:

  2. Search for Christian Schultze-Florey in:

  3. Search for Solaiman Raha in:

  4. Search for Inga Sandrock in:

  5. Search for Melanie Drenker in:

  6. Search for Linda Oberdörfer in:

  7. Search for Annika Reinhardt in:

  8. Search for Inga Ravens in:

  9. Search for Maleen Beck in:

  10. Search for Robert Geffers in:

  11. Search for Constantin von Kaisenberg in:

  12. Search for Michael Heuser in:

  13. Search for Felicitas Thol in:

  14. Search for Arnold Ganser in:

  15. Search for Reinhold Förster in:

  16. Search for Christian Koenecke in:

  17. Search for Immo Prinz in:

Contributions

Sa.R. wrote the manuscript; Sa.R., C.S.-F. and So.R. designed and performed experiments, discussed and analyzed data; I.S., A.R., I.R. and M.B. helped with performing experiments and data analysis; M.D. and L.O. organized, acquired and processed clinical samples; R.G., M.H. and F.T. helped supervise NGS; C.v.K. and A.G. helped supervise clinical sample acquisition; R.F. helped supervise research; C.K. supervised research and the clinical study, and discussed and analyzed data; and I.P. supervised research, discussed and analyzed data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Immo Prinz.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–5 and Supplementary Tables 1 and 2

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ni.3686

Further reading