Session Type: Abstract Submissions (ACR)
Systemic lupus erythematosus (SLE) patients exist an imbalance between CD4+CD25+FoxP3+ T regulatory (Treg) and IL-17-producing cells (Th17). Correction of this Treg/Th17 imbalance may have therapeutic impact for SLE patients. Our previous study demonstrated that bone marrow derived mesenchymal stem cells (MSCs) from SLE patients are defective in immune modulation, which might be involved in Treg/Th17 imbalance, but the mechanisms are not clear yet. As a computer predicted target of miR-663, Transforming growth factor-beta1 (TGF-β1) is an important negative immune regulatory factors secreted by MSCs which could modulate SLE Treg/Th17 imbalance. The aim of this study is to investigate whether miR-663 could contribute to SLE Treg/Th17 imbalance though directly down-regulating TGF-β1 mRNA of SLE BMSCs, to further understand the pathogenesis of SLE.
Methods: BMSCs were isolated, cultured and expanded from iliac crest bone marrow of four healthy controls and five SLE patients. MicroRNA expressions of BMSCs from were determined by MicroRNA array analysis. Real-time PCR was used to further determine the miR-663 expression and TGF-β1 mRNA level in MSCs, secretion of TGF-β1 was detected by ELISA. As a computer predicted target of miR-663, TGF-β1 was determined using the luciferase reporter assay system. MSCs were transfected with miR-663a, pre-miR-663a, and anti-miR-663a, and then co-cultured with PBMCs from SLE patients for 3 days respectively with PHA pre-stimulated, flow cytometry was used to detect their effect on percentage of Treg and Th17 cells.
Mir-663 was significantly up-regulated(2.52- fold higher, P<0.05) in MSCs from SLE patients(n=5) compared to normal controls(n=4) by microRNA array analysis. The expression of miR-663 was further determined by RT-PCR, and the synthesis of TGF-β1 mRNAs was significantly lower(3.88- fold lower, P<0.05) in MSCs from SLE patients. Transfection of SLE MSCs with pre-miR-663a caused significant upregulation of miR-663a expression(8.33- fold higher, n=3, P<0.01) and markedly lower synthesis of TGF-β1 mRNA(1.52-fold lower, n=3, P<0.05), while transfection with anti-miR-663a markedly decreased the miR-663a expression (3.01- fold lower, n=3, P<0.05). The mean value of Treg/Th17 was significantly decreased in PBMCs from SLE patients compared to normal controls (0.48±0.12vs0.65±0.09, n=8, P<0.01). Compared to SLE MSCs, MSCs from normal controls exhibited a better immune suppression effect through up-regulating Treg/Th17 of SLE patients (0.68±0.15vs0.54±0.14, n=3, P<0.05), and transfection of normal MSCs with pre-miR-663a caused significant downregulation of Treg/Th17 compared to miR-663-control group(0.38±0.07vs0.68±0.15, n=3, P<0.01), while transfection with anti-miR-663a led to an opposite effect (0.76±0.08vs0.68±0.15, n=3, P<0.05).
SLE patients exist an imbalance between Treg and Th17 cells, which might be associated with down-regulated TGF-β1 secretion of bone marrow-derived mesenchymal stem cells mediated by miR-663.
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ACR Meeting Abstracts - https://acrabstracts.org/abstract/the-tregth17-imbalance-of-patients-with-systemic-lupus-erythematosus-were-mediated-by-mir-663-through-down-regulating-tgf-%ce%b21-secretion-of-bone-marrow-derived-mesenchymal-stem-cells/