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Iran J Allergy Asthma Immunol. 2015 Aug;14(4):416-26.

In Vitro Generation of IL-35-expressing Human Wharton's Jelly-derived Mesenchymal Stem Cells Using Lentiviral Vector.

Author information

1
Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
2
Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran AND Department of Stem Cells Biology, Stem Cell Technology Research Center, Tehran, Iran.
3
Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
4
Department of Immunology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
5
Department of Stem Cells Biology, Stem Cell Technology Research Center, Tehran, Iran AND National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.

Abstract

Human Wharton's Jelly-derived Mesenchymal Stem Cells (hWJ-MSCs) are easily available cells without transplant rejection problems or ethical concerns compared to bone-marrow-derived MSCs for prospective clinical applications. These cells display immunosuppressive properties and may be able to play an important role in autoimmune disorders. Regulatory T-cells (Treg) are important to prevent autoimmune disease development. Interleukin 35 (IL-35) induces the proliferation of Treg cell populations and reduces the activity of T helper 17 (Th17) and T helper 1 (Th1) cells, which play a central role in initiation of inflammation and autoimmune disease. Recent studies identified IL-35 as a new inhibitory cytokine required for the suppressive function of Treg cells. We created IL-35-producing hWJ-MSCs as a good vehicle for reduction of inflammation and autoimmune diseases. We isolated hWJ-MSCs based on explant culture. HWJ-MSCs were transduced at MOI=50 (Multiplicity of Infection) with lentiviral particles harboring murine Interleukin 35 (mIL-35). Expression of IL-35 in hWJ-MSCs was quantified by an IL-35 ELISA kit. IL-35 bioactivity was analyzed by inhibiting the proliferation of mouse splenocytes using CFSE cell proliferation kit. Frequency of CD4+CD25+CD127 low/neg Foxp3+ Treg cells was measured by flow cytometry. There was an up to 85% GFP positive transduction rate, and the cells successfully released a high level of mIL-35 protein (750 ng/ml). IL-35 managed to inhibit CD4+ T cell proliferation with PHA, and improved the frequency of Treg cells. Our data suggest that transduced hWJ-MSCs overexpressing IL-35 may provide a useful approach for basic research on gene therapy for autoimmune disorders.

KEYWORDS:

IL-35; Lentivirus; Mesenchymal Stem Cells; Vector; Wharton’s Jelly-Derived

PMID:
26547710
[Indexed for MEDLINE]
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