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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. [ABSTRACT FROM AUTHOR]

Asthma is a chronic inflammatory disorder of the airways. The stress is a factor for asthma which indicates a disorder in the function of communicational mediators of nervous and immunological systems such as neurotransmitters. A study indicated that blood serotonin concentration increases in asthmatic patients. Other study indicates that one kind of the serotonin receptors, named 5HT3A, on PBMCs causes secretion of series of pro-inflammatory cytokines which play important roles in allergic asthma disease. Thus, we evaluated the ratio expression level of 5HT3A subtype receptors in asthma. The Peripheral Blood Mononuclear Cells were separated from whole blood of 30 allergic asthmatic patients and 30 normal controls by a gradient density centrifugation technique, then the total cellular RNA was extracted and the cDNA was synthesized. This process was followed by real-time PCR using primer pairs specific for 5-hydroxytryptamine 3A subtype receptor mRNA and beta-actin as internal control. Results revealed that relative gene expression of 5-hydroxytryptamine 3A subtype receptor increased significantly in Peripheral Blood Mononuclear Cells of patients with asthma in comparison with normal individuals. To conclude, considering 5-hydroxytryptamine 3A subtype receptor role in accomplishment of asthma symptoms, this increase in its expression may exacerbate the seriousness of asthma disease.