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Optimization of human Th17 cell differentiation in vitro: evaluating different polarizing factors

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Abstract

Regarding discrepancies that exist among different studies which have tried to clarify critical factors in human Th17 cell differentiation, the aim of this study was to identify the best condition for human Th17 differentiation and to clarify the possible role of TGF-β in differentiation of these cells. Naïve CD4+ T cells were isolated from cord blood samples and cultured either in X-VIVO 15 serum-free medium or RPMI 1640 containing 10% FBS. Purified cells were treated with different combinations of polarizing cytokines (TGF-β, IL-1β, IL-6, IL-23 and IL-21) followed by analysis of the expression of characteristic genes and their relevant cytokines by real-time quantitative RT-PCR and ELISA method, respectively. Our data indicate that a combination of TGF-β plus IL-6 and IL-23 cytokines in X-VIVO 15 serum-free medium could be applied as the best condition for developing human Th17 cells in compare with other studied cytokine treatments. It is shown that TGF-β could be considered as a positive regulator for human Th17 cell differentiation only if applied in average concentrations. Interestingly, polarizing treatments in absence of TGF-β, induced double-secreting Th17 cells which co-express IL-17 and IFN-γ whereas polarization in presence of TGF-β-induced single-secreting (only IL-17 expressing) Th17 cells.

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Acknowledgments

The main part of this work was supplied by Isfahan University of Medical Sciences (Grant No. 187058) and the remaining was financially supported by Iran National Science Foundation (Grant No. 87040290). We wish to thanks Dr. Shahshahani and Mrs Janghorban for their excellent helps in cord blood collection, Mrs Homayouni and Mrs Esmaeili for their kindly collaborations in flow cytometry and ELISA techniques and Mr. Ghoochani for his excellent helps in data interpretation.

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Authors and Affiliations

  1. Immunology Dept., Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    Mazdak Ganjalikhani Hakemi, Alireza Andalib & Abbas Rezaei

  2. Applied Physiology Research Center, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

    Mazdak Ganjalikhani Hakemi & Abbas Rezaei

  3. Biology Dept., Faculty of Sciences, University of Isfahan, Isfahan, Iran

    Kamran Ghaedi

  4. Epidemiology Dept., Faculty of Health, Isfahan University of Medical Sciences, Isfahan, Iran

    Mohsen Hosseini

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  1. Mazdak Ganjalikhani Hakemi
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  2. Kamran Ghaedi
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  4. Mohsen Hosseini
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  5. Abbas Rezaei
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Correspondence to Abbas Rezaei.

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Editor: T. Okamoto

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Ganjalikhani Hakemi, M., Ghaedi, K., Andalib, A. et al. Optimization of human Th17 cell differentiation in vitro: evaluating different polarizing factors. In Vitro Cell.Dev.Biol.-Animal 47, 581–592 (2011). https://doi.org/10.1007/s11626-011-9444-1

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