ABSTRACT
The levels of macrophage migration inhibitory factor (MIF) in peripheral blood mononuclear cell (PBMC) and serum in patients with Hashimoto's thyroiditis (HT) were detected by real-time PCR and ELISA,respectively.The results revealed that the expression of MIF mRNA in PBMC ( Z =-4.276,P＜0.01 ) and protein level in serum ( Z=-5.358,P＜0.01 ) were increased in HT patients,and positively correlated with thyroid specific autoantibodies and TSH levels.
ABSTRACT
[Objective] To assess the role of imbalance between regulatory T (Treg) cells and T helper 17 (Th17) cells in the pathogenesis of atopic dermatitis (AD).[Methods] Peripheral blood was obtained from 41 patients with AD and 38 age- and sex-matched healthy controls.Flow cytometry was performed to determine the percentage of Treg cells (CD4+CD25+Foxp3+ T cells) and Thl7 cells (CD4+ILl7+ T cells),real-time quantitative reverse transcription (RT)-PCR to detect the mRNA expressions of Foxp3 and RORγt,which are the specific transcription factors of Treg and Th17 cells respectively.Serum concentrations of transforming growth factor (TGF)-β,IL-17 and IL-23 were measured by enzyme linked immunosorbent assay(ELISA).Data were statistically assessed by independent-samples t test and Pearson correlation analysis.[Results] The patients with AD showed an obvious decrease in Treg cell percentage,transcription factor Foxp3 mRNA level and Treg/Th17 ratio (2.01％ ± 0.57％ vs.5.04％ ± 1.44％,t =12.47,P＜ 0.01; 0.65 ± 0.19 vs.1.71 ± 0.69,t=9.47,P＜0.01; 1.26 ± 0.61 vs.14.53 ± 5.77,t =14.11,P ＜ 0.01),but a significant increase in peripheral Th17 cell percentage and transcription factor RORγt mRNA level (1.77％ ± 0.55％ vs.0.39％ ± 0.15％,t =14.82,P ＜0.01; 5.97 ± 1.45 vs.1.49 ± 0.57,t =17.78,P ＜ 0.01 ) compared with the healthy controls.Further comparison revealed that Treg/Th17 ratio was significantly lower in patients with acute AD than in those with subacute AD (0.88 ± 0.04 vs.1.29 ± 0.11,t =4.02,P ＜ 0.01 ) and those with chronic AD (2.05 ± 0.24,t =4.83,P ＜ 0.01 ),statistically different between patients with subacute AD and chronic AD (t =2.89,P ＜ 0.05).There was no significant difference in the serum concentration of TGF-β between patients with AD and healthy controls ((15.28 ± 2.34) μg/L vs.(16.56 ± 3.27) μg/L,t =1.96,P＞ 0.05).A significant increase was observed in the serum levels of IL-17 and IL-23 in patients with AD compared with those in the healthy controls( (33.24 ± 7.06)ng/L vs.(11.68 ± 2.67) ng/L,t =17.96,P＜ 0.01; (56.35 ± 12.16) ng/L vs.(18.43 ± 3.90) ng/L,t =18.36,P＜ 0.01).In patients with moderate and severe AD,SCORing atopic dermatitis (SCORAD) index was negatively correlated with the percentage of Treg ceils (r =-0.40,P＜ 0.05 ),but positively correlated with that of Th17 cells (r =0.42,P ＜ 0.05 ).[Conclusion]s There exists a change in Treg/Th 17 ratio,mRN A expressions of RORγt and Foxp3,and serum levels of relevant cytokines in patients with AD,which may lead to immune imbalance and subsequently contribute to the development of AD.
ABSTRACT
Objective To investigate the effects of Trim34α on the activation of luciferase reporter gene containing NF-κB promoter induced by adaptor proteins TAB2. Methods The total RNA was isolated from HeLa cells. After amplification with RT-PCR, the target sequences were cloned into 5'-Flag-pcDNA3.1 (+) vector. The recombinant vector was confirmed by restriction enzyme digestion, colony PCR and sequencing. It was transfected into HEK293T cells to detected Trim34α expression by Western blot. Simultaneously, the effects of Trim34α on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Results Restriction enzyme digestion, colony PCR and sequencing confirmed the vector was constructed successfully, furthermore it expressed Trim34α protein in HEK293T cells. Moreover, trim34α could form high-molecular-weight oligomeric protein, and here we called it trimsome. Interestingly, dual-luciferase assay showed that Trim34α could effectively block TAB2-induced NF-κB activation. Conclusion Trim34α was involved in negative regulation of TAB2-induced NF-κB activation and could form high-molecular-weight oligomer.