title: In Vitro Generation of IL-35-expressing Human Wharton's Jelly-derived Mesenchymal Stem Cells Using Lentiviral Vector.
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.
authors.affiliation: Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Afshin
authors.initials: A
authors.lastname: Amari
authors.affiliation: Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Massoumeh
authors.initials: M
authors.lastname: Ebtekar
authors.affiliation: Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Seyed Mohammad
authors.initials: SM
authors.lastname: Moazzeni
authors.affiliation: 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.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Department of Medical Genetics, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Leila
authors.initials: L
authors.lastname: Mohammadi Amirabad
authors.affiliation: Department of Immunology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
authors.forename: Mohammad Taher
authors.initials: MT
authors.lastname: Tahoori
authors.affiliation: Department of Stem Cells Biology, Stem Cell Technology Research Center, Tehran, Iran AND National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Massumi
mesh_terms: D000818:Animals; D001327:Autoimmune Diseases / Q000628:therapy; D002478:Cells, Cultured; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D007378:Interleukins / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics*; D008213:Lymphocyte Activation; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D050378:T-Lymphocytes, Regulatory / Q000276:immunology; D059631:Wharton Jelly / Q000166:cytology*
chemical_list: D007378:Interleukins; C532417:interleukin-35, human
keywords: IL-35; Lentivirus; Mesenchymal Stem Cells; Vector; Wharton’s Jelly-Derived