Technical Report | Published:

An in vitro system of autologous lymphocytes culture that allows the study of homeostatic proliferation mechanisms in human naive CD4 T-cells

Laboratory Investigationvolume 98pages500511 (2018) | Download Citation

Abstract

The size of peripheral T-cell pool is kept constant throughout life. However, a decline in lymphocyte numbers is a feature of several human disorders, in which fast and slow homeostatic proliferation play a crucial role. Several in vitro and in vivo models have been developed to study such processes. Nevertheless, self- and commensal- antigens, well-known triggers of homeostatic proliferation, have not been examined in these models. We have designed an in vitro culture of human T-cells exposed to rIL7 and autologous antigen-presenting cells (aAPC) that allows the simultaneous characterization of the different types of homeostatic proliferation. Using our model, we first confirmed that both rIL7 and aAPC are survival signals ultimately leading to homeostatic proliferation. In addition, we explored the modulation of different anti-apoptotic, proliferative, activation and homing markers during fast and slow homeostatic proliferation. Finally, different subsets of Treg were generated during homeostatic proliferation in our model. In summary, our in vitro system is able to simultaneously reproduce both types of homeostatic proliferation of human naive CD4 T-cells, and allows the characterization of these processes. Our in vitro system is a useful tool to explore specific features of human homeostatic proliferation in different human lymphopenia-related disorders and could be used as a cell therapy approach.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Goldrath AW, Bevan MJ. Selecting and maintaining a diverse T-cell repertoire. Nat Rev Immunol. 1999;402:255–62.

  2. 2.

    Freitas AA, Rocha B. Population biology of lymphocytes: the flight for survival. Annu Rev Immunol. 2000;18:83–111.

  3. 3.

    Sener A, Tang AL, Farber DL, et al. Memory T-cell predominance following T-cell depletional therapy derives from homeostatic expansion of naive T cells. Am J Transplant. 2009;9:2615–23.

  4. 4.

    Hakim FT, Memon SA, Cepeda R, et al. Age-dependent incidence, time course and consequences of thymic renewal in adults. J Clin Invest. 2005;115:930–9.

  5. 5.

    Takada K, Jameson SC. Naive T cell homeostasis: from awareness of space to a sense of place. Nat Rev Immunol. 2009;9:823–32.

  6. 6.

    Surh CD, Sprent J. Homeostasis of naive and memory T cells. Immunity. 2008;29:848–62.

  7. 7.

    Do J, Foucras G, Kamada N, et al. Both exogenous commensal and endogenous self antigens stimulate T cell proliferation under lymphopenic conditions. Cell Immunol. 2012;272:117–23.

  8. 8.

    Min B, Yamane H, Hu-Li J, et al. Spontaneous and homeostatic proliferation of CD4 T cells are regulated by different mechanisms. J Immunol. 2005;174:6039–44.

  9. 9.

    Ge Q, Rao VP, Cho BK, et al. Dependence of lymphopenia-induced T cell proliferation on the abundance of peptide MHC epitopes and strength of their interaction with T cell receptors. Proc Natl Acad Sci USA. 2001;98:1728–33.

  10. 10.

    Ringhoffer S, Rojewski M, Döhner H, et al. T-cell reconstitution after allogeneic stem cell transplantation: assessment by measurement of the sjTREC /β TREC ratio and thymic naive T cells. Haematologica. 2013;98:1600–8.

  11. 11.

    Williams KM, Hakim FT, Gress RE. T cell immune reconstitution following lymphodepletion. Semin Immunol. 2007;19:318–30.

  12. 12.

    Cimbro R, Vassena L, Arthos J, et al. Interleukin-7 induces expression and activation of integrin α4β7 promoting naive T-cell homing to the intestinal mucosa. Blood. 2012;120:2610–9.

  13. 13.

    Beq S, Nugeyre MT, Ho T, et al. IL-7 Induces immunological improvement in SIV-infected rhesus macaques under antiviral therapy. J Immunol. 2006;176:914–22.

  14. 14.

    Haines CJ, Giffon TD, Lu LS, et al. Human CD4+T cell recent thymic emigrants are identified by protein tyrosine kinase 7 and have reduced immune function. J Exp Med. 2009;206:275–85.

  15. 15.

    Chomont N, El-Far M, Ancuta P, et al. HIV reservoir size and persistence are driven by T cell survival and homeostatic proliferation. Nat Med. 2009;15:893–900.

  16. 16.

    Martinet KZ, Bloquet S, Bourgeois C. Ageing combines CD4 T cell lymphopenia in secondary lymphoid organs and T cell accumulation in gut associated lymphoid tissue. Immun Ageing. 2014;11:1–13.

  17. 17.

    Jones JL, Thompson SA, Loh P, et al. Human autoimmunity after lymphocyte depletion is caused by homeostatic T-cell proliferation. Proc Natl Acad Sci. 2013;110:20200–5.

  18. 18.

    Pearson C, Silva A, Saini M, et al. IL-7 determines the homeostatic fitness of T cells by distinct mechanisms at different signalling thresholds in vivo. Eur J Immunol. 2011;41:3656–66.

  19. 19.

    Hennion-Tscheltzoff O, Leboeuf D, Gauthier SD, et al. TCR triggering modulates the responsiveness and homeostatic proliferation of CD4+thymic emigrants to IL-7 therapy. Blood. 2013;21:4684–93.

  20. 20.

    Booki M, Hidehiro Y, Jane HL, et al. Spontaneous and homeostatic proliferation of CD4 T cells are regulated by different mechanisms. J Immunol. 2005;174:6039–44.

  21. 21.

    Kawabe T, Sun SL, Fujita T, et al. Homeostatic proliferation of naive CD4+T cells in mesenteric lymph nodes generates gut-tropic Th17 cells. J Immunol. 2013;190:5788–98.

  22. 22.

    Sousa AE, Carneiro J, Meier-Schellersheim M, et al. CD4 T cell depletion is linked directly to immune activation in the pathogenesis of HIV-1 and HIV-2 but only indirectly to the viral load. J Immunol. 2002;169:3400–6.

  23. 23.

    Gorfu G, Rivera-Nieves J, Ley K. Role of β7 integrins in intestinal lymphocyte homing and retention. Curr Mol Med. 2009;9:837–50.

  24. 24.

    Rivera-Nieves J, Olson T, Bamias G, et al. L-selectin, alpha 4 beta 1, and alpha 4 beta 7 integrins participate in CD4+T cell recruitment to chronically inflamed small intestine. J Immunol. 2005;174:2343–52.

  25. 25.

    Apostolaki M, Manoloukos M, Roulis M, et al. Role of β7 integrin and the chemokine/chemokine receptor pair CCL25/CCR9 in modeled TNF-dependent Crohn’s disease. Gastroenterology. 2008;134:2025–35.

  26. 26.

    Di Lanni M, Falzetti F, Carotti A, et al. Tregs prevent GVHD and promote immune reconstitution in HLA-haploidentical transplantation. Blood. 2001;117:3921–9.

  27. 27.

    Nguyen VH, Shashidhar S, Chang DS, et al. The impact of regulatory T cells on T-cell immunity following hematopoietic cell transplantation. Blood. 2008;111:945–54.

  28. 28.

    Curotto De Lafaille MA, Lino AC, Kutchukhidze N, et al. CD25- T cells generate CD25+Foxp3+regulatory T cells by peripheral expansion. J Immunol. 2004;173:7259–68.

  29. 29

    Maldonado RA, von Andrian UH. How tolerogenic dendritic cells induce regulatory T cells. Adv Immunol. 2010;108:111–65.

  30. 30.

    Winstead CJ, Fraser JM, Khoruts A. Regulatory CD4+CD25+Foxp3+T cells selectively inhibit the spontaneous form of lymphopenia-induced proliferation of naive T cells. J Immunol. 2008;180:7305–17.

  31. 31.

    Zhang N, Bevan MJ. TGF-beta signaling to T cells inhibits autoimmunity during lymphopenia-driven proliferation. Nat Immunol. 2012;13:667–73.

  32. 32.

    Surh CD, Sprent J. TGF-β puts the brakes on homeostatic proliferation. Nat Immunol. 2012;13:628–30.

  33. 33.

    Matsuoka K, Kim HT, McDonough S, et al. Altered regulatory T cell homeostasis in patients with CD4+lymphopenia following allogeneic hematopoietic stem cell transplantation. J Clin Invest. 2010;120:1479–93.

  34. 34.

    Reichardt P, Dornbach B, Gunzer M. APC, T cells, and the immune synapse. Curr Top Microbiol Immunol. 2010;340:229–49.

Download references

Acknowledgements

This work was supported by grants from the Instituto de Salud Carlos III, Fondo de Investigación Sanitaria (FIS; PI14/01693) (Co-funded by European Regional Development Fund/European Social Fund) “Investing in your future”) and the Junta de Andalucía, Consejería de Economía, Innovación, Ciencia y Empleo (Proyecto de Investigación de Excelencia; CTS2593). The Spanish AIDS Research Network of Excellence supported this study (RIS; RD12/0017/0029 and RD16/0025/0019). Y.M. Pacheco was supported by the Fondo de Investigación Sanitaria through the “Miguel Servet” program (CPII13/00037), and by the Consejería de Salud y Bienestar Social of Junta de Andalucía through the “Nicolás Monardes” program (C-0010/13). We thank Mª Antonia Abad, Marta de Luna and Cytometry Service of IBiS, especially Mª José Castro, for their technical assistance. We also thank to Manuel Moyano from Centro Regional de Transfusión Sanguínea de Sevilla-Huelva y Banco de Tejidos (Seville, Spain) for the kind gift of samples.

Author information

Affiliations

  1. Laboratory of Immunovirology, Institute of Biomedicine of Seville, IBiS, Virgen del Rocío University Hospital/CSIC/University, Seville, 41013, Spain

    • Isaac Rosado-Sánchez
    • , Amaia González-Magaña
    • , María M Pozo-Balado
    • , Inés Herrero-Fernández
    • , María J Polaino
    • , María M Rodríguez-Méndez
    • , Manuel Leal
    •  & Yolanda M Pacheco
  2. Immunology Service, Institute of Biomedicine of Seville, IBiS, Virgen del Rocío University Hospital/CSIC/University, Seville, 41013, Spain

    • María Francisca González-Escribano

Authors

  1. Search for Isaac Rosado-Sánchez in:

  2. Search for Amaia González-Magaña in:

  3. Search for María M Pozo-Balado in:

  4. Search for Inés Herrero-Fernández in:

  5. Search for María J Polaino in:

  6. Search for María M Rodríguez-Méndez in:

  7. Search for María Francisca González-Escribano in:

  8. Search for Manuel Leal in:

  9. Search for Yolanda M Pacheco in:

Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding authors

Correspondence to Isaac Rosado-Sánchez or Yolanda M Pacheco.

About this article

Publication history

Received

Revised

Accepted

Published

DOI

https://doi.org/10.1038/s41374-017-0006-3