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In vitro treatment of murine splenocytes with extracellular vesicles derived from mesenchymal stem cells altered the mRNA levels of the master regulator genes of T helper cell subsets

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Abstract

Introduction

The purpose of the current study was to evaluate the effect of mesenchymal stem cells-derived extracellular vesicles (MSC-EVs) on the production of cytokines and expression of genes, which are corresponded to the subsets of T helper cells.

Materials and Methods

The supernatant of the second passage of MSCs that had been isolated from C57BL/6 mice abdominal adipose tissue was used to collect the MSC-EV. Splenocytes of healthy mice were activated using anti-CD3 and anti-CD28 antibodies and simultaneously were treated using the MSC-EVs. The proliferation rate of lymphocytes and the frequency of regulatory T cells were measured using flow cytometry. In addition, the expressions of T helper cell subset-specific transcription factors were evaluated using a real-time PCR assay. To appraise the effects of MSC-EV on splenocytes, the levels of IFN-γ, IL-17A, IL-10, and TGF-β were measured using ELISA.

Results

The results showed that the treatment of the CD3/CD28-activated splenocytes with MSC-EV did not statistically change the proliferation of CD3+ splenocytes. However, after the treatment, the mRNA levels of Foxp3 and Elf4 as well as the frequency of regulatory T cells was significantly higher when compared to the control group. The expression levels of Gata3, Rorc, and Tbx21 were down-regulated while, the corresponding cytokines levels did not alter.

Conclusion

The results revealed that the in vitro treatment of MSC-EV was associated with the increase in the frequency of CD4+CD25+FOXP3+ T cells and upregulation of Foxp3 mRNA level.

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Acknowledgements

The present study was supported financially by grants from the deputy of research, Shahid Beheshti University of Medical Sciences (Grant Nos. 8818 and 19547).

Funding

Funding was provided by Shahid Beheshti University of Medical Sciences (Grant Nos. 8818 and 19547).

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Authors and Affiliations

  1. Department of Biology, Faculty of science, University of Guilan, Rasht, Iran

    Alireza Yeganeh

  2. Department of Medical Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Anwar Fathollahi, Seyed Mahmoud Hashemi & Farshid Yeganeh

  3. Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Farshid Yeganeh

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  1. Alireza Yeganeh
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  2. Anwar Fathollahi
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  3. Seyed Mahmoud Hashemi
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  4. Farshid Yeganeh
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Correspondence to Farshid Yeganeh.

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Yeganeh, A., Fathollahi, A., Hashemi, S.M. et al. In vitro treatment of murine splenocytes with extracellular vesicles derived from mesenchymal stem cells altered the mRNA levels of the master regulator genes of T helper cell subsets. Mol Biol Rep 50, 3309–3316 (2023). https://doi.org/10.1007/s11033-023-08247-1

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  • DOI: https://doi.org/10.1007/s11033-023-08247-1

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