ABSTRACT
ObjectiveTo investigate the effect of HBsAg on the expression of interferon-α (IFN-α) in peripheral blood plasmacytoid dendritic cells (pDCs) induced by the stimulator of interferon genes (STING) signaling pathway activated by cyclic GMP-AMP (cGAMP). MethodPeripheral venous blood was collected from healthy adults and the patients with chronic hepatitis B virus (HBV) infection who attended the outpatient service or were hospitalized in Department of Infectious Diseases, The Affiliated Hospital of Xuzhou Medical University, from February to December 2016, and peripheral blood mononuclear cells (PBMCs) were isolated and extracted. After the STING agonist cGAMP was added to PBMCs, ELISA was used to measure the levels of IFN-α, interferon-β, and tumor necrosis factor-α in supernatant. PBMCs from healthy adults were pre-incubated with HBsAg and then stimulated by cGAMP, and supernatant was collected to measure IFN-α. The magnetic-activated cell sorting method was used to remove pDCs from PBMCs, and after culture with cGAMP, ELISA was used to measure the level of IFN-α in supernatant. PBMCs from healthy adults were stimulated by HBsAg and/or cGAMP, and then flow cytometry was used to measure the frequency of pDCs. The independent samples t-test was used for comparison of continuous data between two groups. ResultsPBMCs from the patients with chronic HBV infection stimulated by cGAMP in vitro had a significantly lower level of IFN-α than healthy controls (469.72±18.95 vs 599.90±84.06, t=4.868, P=0.001). PBMCs from healthy adults co-cultured with HBsAg and stimulated by cGAMP had a significantly lower level of IFN-α than those in the non-HBsAg group (448.5±52.0 vs 571.0±30.8, t=4.500, P=0.011). Compared with PBMCs containing pDCs, PBMCs without pDCs stimulated by cGAMP had a significant reduction in the level of IFN-α (164.50±40.73 vs 339.50±35.33, t=6.482, P=0.001). Compared with PBMCs from healthy adults stimulated by cGAMP, PBMCs pre-incubated with HBsAg and then stimulated by cGAMP had a significant reduction in the frequency of pDCs (0.12%±0.04% vs 0.24%±0.04%, t=5.176, P=0.014). ConclusionHBsAg can inhibit the expression of IFN-α induced by the STING pathway in pDCs activated by cGAMP.
ABSTRACT
Objective To investigate the effects RORrt (RORrt),IRF8 (IRF8) and STAT3 (STAT3) in peripheral blood CD4+T cells on the cell proliferation and differentiation in elderly patients with iron-overload myelodysplastic syndrome(MDS).Methods A prospective case-control study was conducted.Twenty-two elderly hospitalized patients(12 males and 10 females)aged 60-78 years with iron-overload MDS from Jan.2017 to Dec.2018 were enrolled and considered as the observation group.Twenty MDS patients without iron overload hospitalized in the same period were selected as the non-iron overload group,and 26 healthy elderly people were considered as the healthy control group.Peripheral blood monocytes(PBM)were prepared and resident CD4+T cells were sorted by flow cytometry.The mRNA and protein expression levels of transcription factors of RORrt,p-STAT3 and IRF8 were detected by quantitative real time polymerase chain reaction(qRT-PCR)and Western blotting.Results In peripheral blood CD4+T cells,the mRNA expression level of RORrt and p-STAT3 were higher and that of IRF8 was lower in the iron-overload group than in the non-iron overload group and the healthy control group(42.634± 18.613 vs.21.289 ± 15.158 and 22.520 ± 9.896;29.710±9.689 vs.12.355±4.681 and 9.818±3.845;19.293±8.258 vs.23.785±12.498 and 69.619±23.768,P＜0.01).In peripheral blood CD4+T cells,the protein expression level of RORrt and p-STAT3 was higher,and that of IRF8 was lower in the iron overload group than in the non-iron overload group and healthy control group(P＜0.01).Conclusions The abnormalities of the mRNA and protein expression levels of transcription factors of RORrt,IRF8 and p-STAT3 in CD4+ T cells play a fundamental role in the pathogenesis of iron overload MDS in elderly patients.
ABSTRACT
Objective To evaluate the effects of pulsed radiofrequency (PRF) application to dorsal root ganglia on the expression of interferon regulatory factor 8 (IRF8) in the spinal cord and brain-derived neurotrophic factor (BDNF) protein in the nucleus accumbens of rats with neuropathic pain (NP).Methods Forty healthy pathogen-free male Wistar rats,weighing 180-200 g,aged 2 months,were divided into 4 groups (n=10 each) using a random number table:sham operation group (group Sham),group NP,sham PRF group (group SPRF) and PRF group.NP was induced by chronic constriction injury (CCI) to the left sciatic nerve of anesthetized rats.The mechanical paw withdrawal threshold (MWT) was measured before CCI and at 3,7,10,14,21,28,35 and 42 days after CCI.Sucrose preference test and forced-swim test were performed at 42 days after CCI for determination of the expression of IRF8 in the spinal cord and BDNF in the nucleus accumbens by Western blot.Results Compared with group Sham,the MWT at each time point after CCI and rate of preference for sucrose were significantly decreased,the duration of immobility in forced-swim test was prolonged,and the expression of IRF8 and BDNF was up-regulated in NP,SPRF and PRF groups (P＜0.05).Compared with group NP,the MWT at 10-42 days after CCI and rate of preference for sucrose were significantly increased,the duration of immobility in forced-swim test was shortened,and the expression of IRF8 and BDNF was down-regulated in group PRF (P＜0.05),and no significant changes were found in the parameters mentioned above in group SPRF (P＞O.05).Conclusion The mechanism by which PRF application to dorsal root ganglia alleviates NP and depressive-like behaviors is probably related to down-regulation of the expression of IRF8 in the spinal cord and BDNF in the nucleus accumbens of rats.