RESUMEN
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 2×10[5] 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 2-3×10[7] 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
RESUMEN
Unrestricted somatic stem cells [USSC] are cord blood stem cells that have been considered as candidates for the regulation of immune responses. Therefore, potential exists for their use in the suppression of immune response after transplantation surgery. The aim of this study was evaluation of the effect of USSC on mixed lymphocyte reaction [MLR] as a model for graft rejection. USSC and mesanchymal stem cells [MSC] were isolated and cultured from cord blood and bone morrow, respectively. The immunophenotypes of USSC and MSC were evaluated by flow cytometery and USSC and MSC were cocultured with peripheral blood lymphocytes [PBL] in an MLR to evaluate the immunomodulatory effect of these cells as a percentage of the control response. Current study demonstrated that proliferation of lymphocytes in the MLR was decreased after treatment with USSC, in a similar fashion to that seen with MSC. It can be concluded that USSC have similar regulatory effects as MSC on the MLR, which can be used as an indicator for potential organ rejection after transplantation. Therefore, the immunoregulatory effect of these cells could be used in the clinic during organ transplantation and in the management of autoimmunity