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. 2017 Nov;14(5):4807-4812.
doi: 10.3892/etm.2017.5119. Epub 2017 Sep 15.

Role of miR-124a in T cell activation and immunity in AIDS patients

Wenge Zhao  1 , Chuansheng Liu  2 , Changhe Shi  3 , Tianli Fan  1 , Kaiqiu Chu  4 , Yanli Ma  1
Affiliations

Role of miR-124a in T cell activation and immunity in AIDS patients

Wenge Zhao et al. Exp Ther Med. 2017 Nov.

Abstract

The role of microRNA-124a (miR-124a) in the regulation of T cell activation and immunity in patients with AIDS, was studied to provide new insights for the study, diagnosis, alleviation and treatment of AIDS. RT-qPCR technique was used to quantitatively analyze the expression of miR-124a in peripheral blood CD4+ T cells. Dual-luciferase reporter assay system was established to report possible regulatory relations between miR-124a and its potential target gene SIRT1. RT-qPCR and western blot analysis were used to detect the expression level of mRNA and protein of the target genes in T cells. Normal CD4+ T cells from controls were transfected with miR-124a mimics and its negative control, and miR-124a inhibitor and its negative control were transfected into CD4+ T cells from patients with AIDS by T lymphocyte transfection kit to detect the relative expression level of SIRT1 mRNA and protein. The levels of interferon (IFN)-γ, interleukin (IL)-10, transforming growth factor (TGF)-β, IL-2, IL-4 and IL-6 secreted by T helper cells were detected by enzyme-linked immunosorbent assay (ELISA). miR-124a was upregulated in CD4+ T cells of patients with AIDS. The results of firefly luciferase activity detection showed that miR-124a can directly interact with target gene SIRT1 and negatively regulate its expression. miR-124a mimics/inhibitor transfection experiments showed that overexpression of miR-124a in normal CD4+ T cells significantly reduced SIRT1 expression compared with control group, and the expression of miR-124a was positively correlated with IL-10 and TGF-β expression and negatively correlated with IFN-γ expression, but showed no correlation with other cytokines. In AIDS patients, the inhibition of expression of miR-124a in CD4+ T cells significantly increased the expression of SIRT, at the same time, the expression levels of IL-10 and TGF-β were significantly decreased, while the expression level of IFN-γ was significantly increased and no significant difference was found in the expression of other cytokines. The expression of miR-124a in CD4+ T cells of AIDS patients was upregulated and the Th2 type CD4+ T cells are activated by SIRT1 expression inhibition, which in turn enhance the immunity of HIV-infected cells. Our study provides a new molecular target for the diagnosis, alleviation and treatment of AIDS.

Keywords: AIDS; SIRT1; T cell activation; immunity; miR-124a.

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Figures

Figure 1.
Figure 1.
The relative expression levels of miR-124a in CD4+ T cells of two groups of people. *p<0.05.
Figure 2.
Figure 2.
Luciferase relative activities after transfection with different reporter vectors and miR-124a. *p<0.05.
Figure 3.
Figure 3.
miR-124a regulates the expression of target gene SIRT1. (A) Comparison of RT-qPCR results for SIRT1 mRNA levels in CD4+ T cells of AIDS patients and healthy controls. (B) Comparison of SIRT1 protein levels in CD4+ T cells of patients and healthy people. (C) The relative expression levels of miR-124a and SIRT1 in AIDS patients. *p<0.05.
Figure 4.
Figure 4.
The effects of miR-124a overexpression on the expression levels of SIRT1 mRNA and protein and cytokine. *p<0.05.
Figure 5.
Figure 5.
The effect of miR-124a expression inhibition on the expression levels of SIRT1 mRNA and protein and related cytokine proteins. *p<0.05.
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