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 (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