Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice Model

Objective: Immunotherapy and gene therapy play important roles in modern medicine. The aim of this study is to evaluate the overexpression of interleukin-4 (IL-4), IL-10 and leukemia inhibitory factor (LIF) in Wharton’s jelly stem cells (WJSCs) in the experimental autoimmune encephalomyelitis (EA...

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Main Authors: Ahmad Hosseini, Hajar Estiri, Haleh Akhavan Niaki, Akram Alizadeh, Baharak Abdolhossein Zadeh, Sayyed Mohammad Hossein Ghaderian, Akbar Farjadfar, Ali Fallah
Format: Article
Language:English
Published: Royan Institute (ACECR), Tehran 2017-08-01
Series:Cell Journal
Subjects:
Online Access:http://celljournal.org/journal/article/14336/download
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author Ahmad Hosseini
Hajar Estiri
Haleh Akhavan Niaki
Akram Alizadeh
Baharak Abdolhossein Zadeh
Sayyed Mohammad Hossein Ghaderian
Akbar Farjadfar
Ali Fallah
author_facet Ahmad Hosseini
Hajar Estiri
Haleh Akhavan Niaki
Akram Alizadeh
Baharak Abdolhossein Zadeh
Sayyed Mohammad Hossein Ghaderian
Akbar Farjadfar
Ali Fallah
author_sort Ahmad Hosseini
collection DOAJ
description Objective: Immunotherapy and gene therapy play important roles in modern medicine. The aim of this study is to evaluate the overexpression of interleukin-4 (IL-4), IL-10 and leukemia inhibitory factor (LIF) in Wharton’s jelly stem cells (WJSCs) in the experimental autoimmune encephalomyelitis (EAE) mice model. Materials and Methods: In this experimental study, a DNA construction containing IL- 4, IL-10 and LIF was assembled to make a polycistronic vector (as the transfer vector). Transfer and control vectors were co-transfected into Human Embryonic Kidney 293 (HEK-293T) cells with helper plasmids which produced recombinant lentiviral viruses (rLV). WJSCs were transduced with rLV to make recombinant WJSC (rWJSC). In vitro protein and mRNA overexpression of IL-4, LIF, and IL-10 were evaluated using quantitative polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA) and western blot (WB) analysis. EAE was induced in mice by MOG-CFA and pertussis toxin. EAE mice were injected twice with 2×105 rWJSCs. The in vivo level of IL-4, LIF, IL-10 cytokines and IL-17 were measured by ELISA. Brain tissues were analyzed histologically for evaluation of EAE lesions. Results: Isolated WJSCs were performed to characterize by in vitro differentiation and surface markers were analyzed by flow cytometry method. Cloning of a single lentiviral vector with five genes was done successfully. Transfection of transfer and control vectors were processed based on CaPO4 method with >90% efficiency. Recombinant viruses were produced and results of titration showed 2-3×107 infection-unit/ml. WJSCs were transduced using recombinant viruses. IL-4, IL-10 and LIF overexpression were confirmed by ELISA, WB and qPCR. The EAE mice treated with rWJSC showed reduction of Il-17, and brain lesions as well as brain cellular infiltration, in vivo. Weights and physical activity were improved in gene-treated group. Conclusion: These results showed that gene therapy using anti-inflammatory cytokines can be a promising approach against multiple sclerosis (MS). In addition, considering the immunomodulatory potential of WJSCs, an approach using a combination of WJSCs and gene therapy will enhance the treatment efficacy.
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spelling doaj.art-dba64883279e46a48862990a500fe4a32022-12-21T21:51:50ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142017-08-0119336137410.22074/cellj.2017.4497Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice ModelAhmad Hosseini0Hajar Estiri1Haleh Akhavan Niaki2Akram Alizadeh3Baharak Abdolhossein Zadeh4Sayyed Mohammad Hossein Ghaderian5Akbar Farjadfar6Ali Fallah7Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, IranCellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, IranCellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, IranDepartment of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, IranUrogenital Stem Cell Research, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Biotechnology, Fasa University of Medical Sciences, Fasa, IranBioViva USA Inc, Bainbridge Island WA, USAObjective: Immunotherapy and gene therapy play important roles in modern medicine. The aim of this study is to evaluate the overexpression of interleukin-4 (IL-4), IL-10 and leukemia inhibitory factor (LIF) in Wharton’s jelly stem cells (WJSCs) in the experimental autoimmune encephalomyelitis (EAE) mice model. Materials and Methods: In this experimental study, a DNA construction containing IL- 4, IL-10 and LIF was assembled to make a polycistronic vector (as the transfer vector). Transfer and control vectors were co-transfected into Human Embryonic Kidney 293 (HEK-293T) cells with helper plasmids which produced recombinant lentiviral viruses (rLV). WJSCs were transduced with rLV to make recombinant WJSC (rWJSC). In vitro protein and mRNA overexpression of IL-4, LIF, and IL-10 were evaluated using quantitative polymerase chain reaction (qPCR), enzyme-linked immunosorbent assay (ELISA) and western blot (WB) analysis. EAE was induced in mice by MOG-CFA and pertussis toxin. EAE mice were injected twice with 2×105 rWJSCs. The in vivo level of IL-4, LIF, IL-10 cytokines and IL-17 were measured by ELISA. Brain tissues were analyzed histologically for evaluation of EAE lesions. Results: Isolated WJSCs were performed to characterize by in vitro differentiation and surface markers were analyzed by flow cytometry method. Cloning of a single lentiviral vector with five genes was done successfully. Transfection of transfer and control vectors were processed based on CaPO4 method with >90% efficiency. Recombinant viruses were produced and results of titration showed 2-3×107 infection-unit/ml. WJSCs were transduced using recombinant viruses. IL-4, IL-10 and LIF overexpression were confirmed by ELISA, WB and qPCR. The EAE mice treated with rWJSC showed reduction of Il-17, and brain lesions as well as brain cellular infiltration, in vivo. Weights and physical activity were improved in gene-treated group. Conclusion: These results showed that gene therapy using anti-inflammatory cytokines can be a promising approach against multiple sclerosis (MS). In addition, considering the immunomodulatory potential of WJSCs, an approach using a combination of WJSCs and gene therapy will enhance the treatment efficacy.http://celljournal.org/journal/article/14336/downloadGene TherapyMultiple SclerosisWharton’s Jelly Stem CellsCytokines
spellingShingle Ahmad Hosseini
Hajar Estiri
Haleh Akhavan Niaki
Akram Alizadeh
Baharak Abdolhossein Zadeh
Sayyed Mohammad Hossein Ghaderian
Akbar Farjadfar
Ali Fallah
Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice Model
Cell Journal
Gene Therapy
Multiple Sclerosis
Wharton’s Jelly Stem Cells
Cytokines
title Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice Model
title_full Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice Model
title_fullStr Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice Model
title_full_unstemmed Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice Model
title_short Multiple Sclerosis Gene Therapy Using Recombinant Viral Vectors: Overexpression of IL-4, IL-10 and Leukemia Inhibitory Factor in Wharton’s Jelly Stem Cells in The EAE Mice Model
title_sort multiple sclerosis gene therapy using recombinant viral vectors overexpression of il 4 il 10 and leukemia inhibitory factor in wharton s jelly stem cells in the eae mice model
topic Gene Therapy
Multiple Sclerosis
Wharton’s Jelly Stem Cells
Cytokines
url http://celljournal.org/journal/article/14336/download
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