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

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HOSSEINI AHMAD; ESTIRI HAJAR; AKHAVAN NIAKI HALEH; ALIZADEH AKRAM; ABDOLHOSSEIN ZADEH BAHARAK; GHADERIAN SAYYED MOHAMMAD HOSSEIN; FARJADFAR AKBAR; FALLAH ALI

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Journal: Cell Journal (Yakhteh) Year:2017 | Volume:19 | Issue:3 Page(s): 361-374

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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

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Keywords

GENE THERAPY

MULTIPLE SCLEROSIS

WHARTON’S JELLY STEM CELLS

CYTOKINES

Abstract

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-10and 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 andIL-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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Biosimilar Gene Therapy: Investigational Assessment of Secukinumab Gene Therapy

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APA: Copy

HOSSEINI, AHMAD, ESTIRI, HAJAR, AKHAVAN NIAKI, HALEH, ALIZADEH, AKRAM, ABDOLHOSSEIN ZADEH, BAHARAK, GHADERIAN, SAYYED MOHAMMAD HOSSEIN, FARJADFAR, AKBAR, & FALLAH, ALI. (2017). 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 (YAKHTEH), 19(3), 361-374. SID. https://sid.ir/paper/691533/en

Vancouver: Copy

HOSSEINI AHMAD, ESTIRI HAJAR, AKHAVAN NIAKI HALEH, ALIZADEH AKRAM, ABDOLHOSSEIN ZADEH BAHARAK, GHADERIAN SAYYED MOHAMMAD HOSSEIN, FARJADFAR AKBAR, FALLAH ALI. 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 (YAKHTEH)[Internet]. 2017;19(3):361-374. Available from: https://sid.ir/paper/691533/en

IEEE: Copy

AHMAD HOSSEINI, HAJAR ESTIRI, HALEH AKHAVAN NIAKI, AKRAM ALIZADEH, BAHARAK ABDOLHOSSEIN ZADEH, SAYYED MOHAMMAD HOSSEIN GHADERIAN, AKBAR FARJADFAR, and 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 (YAKHTEH), vol. 19, no. 3, pp. 361–374, 2017, [Online]. Available: https://sid.ir/paper/691533/en

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