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Objective:To investigate the regulatory effects of nuclear factor-κB (NF-κB) on dendritic cell (DC) maturation and function through solute carrier family 1 member 2 (Slc1a2).Methods:Mouse bone marrow-derived DCs were transfected with Slc1a2-specific siRNA and an overexpression Slc1a2 eukaryotic expression vector. The real-time fluorescence quantitation (RT-PCR) and Western Blot methods were used to detect knockdown and overexpression efficiency. The expression of surface molecules (CD40, CD80) and major histocompatibility complex Ⅱ (MHCⅡ) of DCs was detected by flow cytometry. ELISA was used to detect the secretion of the cytokines interleukin (IL)-12, IL-6, and transforming growth factor-β (TGF-β). The effects of knockdown of Slc1a2 on DC maturation and function and the effects of overexpression of Slc1a2 on DC maturation and function were reflected by the above assay results. A mixed lymphocyte culture assay was used to investigate the effect of Slc1a2 on T cell proliferation, and an ELISA was used to detect the lavel of IL-17A. Changes in the relative fluorescence intensity of FITC in DCs were analyzed by flow cytometry to investigate the ability of Slc1a2 overexpression on antigen phagocytosis. Finally, DCs were pretreated with an NF-κB inhibitor, toluoylphenylalanine chloromethyl ketone (TPCK), and the effect of TPCK on the expression of Slc1a2 in DCs and DC maturation was examined.Results:Slc1a2 expression was found to be high in DC treated with lipopolysaccharides (LPS) ( P<0.001). The knockdown of Slc1a2 decreased DC maturation and ability to stimulate the proliferation of CD4 + T cells ( P<0.001) and inhibited IL-17 secretion ( P<0.01). Overexpression of Slc1a2 promoted DC maturation and ability to stimulate the proliferation of CD4 + T cells(all P<0.01) Pretreatment of DC with the NF-κB inhibitor TPCK inhibited the expression of Slc1a2 at mRNA and protein levels induced by LPS. Conclusions:NF-κB regulates Slc1a2 expression, which affects the maturation and function of DC.
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Dendritic cells (DC) as professional antigen-presenting cells can activate na?ve T cells and play an important role in bridging innate immunity and acquired immunity. Therefore, the differentiation and maturation of DC and the mechanism in its function regulatory have attracted much attention. With the rapid development of high-throughput sequencing, noncoding RNAs (ncRNAs) have been gradually known. In recent years, increasing studies have reported that ncRNAs play an importantly role in regulating the differentiation and maturation of DC. In this paper, the regulatory effects of microRNAs and long ncRNAs on the differentiation, maturation and function of DC were reviewed.
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Objective To investigate the effects of Glycyrrhiza uralensis Fisch L. crude polysac-charides (GUCP) as an adjuvant on the immunity of mice and the immune responses induced by human pap-illoma virus ( HPV)-DNA vaccine. Methods ICR mice were injected with different concentrations of GUCP by different ways to detect the influences of GUCP on body weight, organ indexes and the numbers of immune cells in spleen. C57BL/ 6 mice were co-immunized with HPV-DNA vaccine and GUCP to detect the adjuvant efficacy on antigen-specific cellular and humoral immune responses. Results GUCP in all injected groups had no side effect on mouse body weight and liver, heart, lung and kidney indexes, but intraperitone-al injection of GUCP significantly increased spleen and thymus indexes and the numbers of B cells, CD4+ T cells, macrophages and dendritic cells in spleen. Subcutaneous injection of GUCP significantly increased the numbers of B cells and macrophages in spleen and intragastric administration significantly increased the num-bers of CD4+ and CD8+ T cells in spleen. Furthermore, GUCP as an adjuvant enhanced the antigen-specific CD4+ and CD8+ T cell responses and the levels of IgG, IgG1 and IgG2a induced by HPV-DNA vaccine at a certain degree. Conclusion GUCP enhanced the immunity of mice and the antigen-specific cellular and hu-moral immune responses induced by HPV-DNA vaccine. These results suggested that GUCP might be used as an adjuvant for DNA vaccine.
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Objective@#As solitary fibrous tumor (SFT) and hemangiopericytoma (HPC) share the same molecular genetics features, the 2016 WHO classification of central nervous system (CNS) tumors had created the combined term SFT/HPC and assigns three grades. This study aims to investigate the clinicopathologic characteristics, diagnosis, differential diagnosis and prognosis of CNS SFT/HPC.@*Methods@#Seventy-one cases of CNS SFT and HPC were retrospectively reclassified and studied. Histopathological, immunohistochemical and imaging features were analyzed. The follow-up data were analyzed.@*Results@#There were 37 male and 34 female patients. The median age was 48 years (range, 3-77 years). Twelve cases (17%) were WHO grade Ⅰ, 26 (37%) were WHO grade Ⅱ and 33 (46%) were WHO grade Ⅲ. Microscopically the tumor could show traditional SFT phenotype, HPC phenotype or mixed phenotype. Immunochemically, 97%(69/71) were positive for STAT6, with 96%(66/69)showing diffuse strong staining. Approximately 90% were diffusely positive for bcl-2, CD99 and vimentin. The expression rate of CD34 decreased with increasing tumor grade, and the mean expression rate was 78%. SSTR2a was variably expressed in 10% (7/71) of cases including one case showing strong cytoplasmic staining. A few cases expressed EMA, CD57 and S-100 focally. The Ki-67 index ranged from 1% to 50%. Thirty four patients were followed up for 8-130 months; 12 patients(35%)had recurrences, and two (6%) had liver metastases.@*Conclusions@#CNS SFT/HPC is relatively uncommon. There was significant morphological overlap or transition between different grades. STAT6 is a specific marker for the diagnosis of this tumor. Surgical resection is the preferred treatment. WHO grade Ⅱ and Ⅲ SFT/HPC show rates of local recurrence and systemic metastasis, with liver being the most common site of extracranial metastasis.
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Objective To investigate the effects of water extract of Glycyrrhiza uralensis Fisch.(GUWE) on the activation of RAW264.7 cells and the possible mechanism.Methods RAW264.7 cells were treated with GUWE containing different concentrations of polysaccharide (10, 50, 100, 500 μg/ml).Viability of these cells was analyzed by MTT assay.Phagocytic activity and surface molecules expressed on these cells were detected by flow cytometry.Levels of cytokines were analyzed by ELISA.Western blot assay was performed to analyze the activation of key molecules in TLR4 signaling pathway.Results GUWE at the concentration of 500 μg/ml significantly decreased the viability of RAW264.7 cells, but significantly increased the viability of RAW264.7 cells at concentrations of 50 μg/ml and 100 μg/ml.GUWE significantly enhanced the phagocytic activity of RAW264.7 cells as well as the expression of cytokines and costimulatory molecules in a dose-dependent manner.Further analysis indicated that the activation of RAW264.7 cells induced by GUWE was suppressed by TLR4 inhibitor.Moreover, GUWE enhanced the phosphorylation of NF-kB p65 and TLR4 downstream mitogen-activated protein kinases (MAPKs) including p38, extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK).Conclusion This study indicates that GUWE promotes the activation of RAW264.7 cells through TLR4 signaling pathway.