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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


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.

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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.

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  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


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The authors declare that they have no conflict of interest.

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Correspondence to Isaac Rosado-Sánchez or Yolanda M Pacheco.

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