Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology最新文献

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease involving multiple tissues and organs. Frequent flare has been regarded as one of the main problems in the diagnosis and treatment of SLE, while the causes for frequent flare has remained to be unclear. We summarized the survival status and the situation of recurrence of SLE patients. The research progress regarding the recurrence and its mechanism will be reviewed from the aspects of genetic factors, environmental factors (i.e. infection, ultraviolet, vitamin D, chemical pollutants), drug and patient compliance, systemic damage and disease status, sex hormones and pregnancy, and social psychology. The implication is to explore effective clinical intervention to alleviate the SLE disease and improve the living quality of the patients.

Objective To investigate the effect of interleukin-34 (IL-34) on the odontogenic and osteogenic differentiation of stem cells from the apical papilla (SCAPs) in rats. Methods SCAPs were isolated and cultured by enzyme digestion method, and the expression of IL-34 in SCAPs was detected by real-time fluorescence quantitative PCR(RT-PCR). MTT assay was used to analyze the effects of different concentrations of IL-34 on SCAPs' proliferation in rats. The mineralization was observed by alizarin red staining, and the proliferation capacity was detected by scratch test. The expressions of alkaline phosphatase (ALP), dentin sialophosphoprotein (DSPP), runt-related transcription factor 2 (Runx2) and critical transcription factor osterix (OSX) were detected by RT-PCR. The protein expressions of ALP, DSPP, Runx2 and OSX were detected by Western blot analysis. Results The maximum concentration of IL-34 promoting the proliferation of SCAPs in rats was 100 ng/mL. Aalizarin red staining showed that IL-34 could promote the mineralization of SCAPs. RT-PCR and Western blot analysis showed that 100 ng/mL IL-34 could promote the expression of ALP, DSPP, Runx2 and OSX. Conclusion IL-34 can promote the proliferation and odontogenic/osteogenic differentiation of SCAPs in rats.

Objective To investigate the effect of calcium voltage gated channel subunit α 1C antisense RNA2 (CACNA1C-AS2) on malignant biological characteristics of esophageal cancer cells by regulating epithelial mesenchymal transition (EMT). Methods CACNA1C-AS2 expression levels in paracancerous tissues and esophageal cancer tissues were analyzed by TCGA database. Real-time quantitative PCR was used to detect the expression of CACNA1C-AS2 mRNA in esophageal cancer cells. Following the knockdown and high expression of CACNA1C-AS2 in esophageal cancer cells, the viability of the cells was tested by MTT assay and cell colony formation assay. Transwell^TM chamber method was used to measure the invasion and longitudinal migration of the cells. The horizontal migration ability of the cells was evaluated by wound healing test. The apoptosis rates of cells were detected by flow cytometry. Western blot analysis was used to detect the expressions of N-cadherin, vimentin and slug. Results CACNA1C-AS2 expression levels were low in esophageal cancer tissues and cell lines. After knocking down the expression of CACNA1C-AS2 in EC-9706 cells and Eca-109 cells, the ability of invasion and migration and viability of esophageal cancer cells were significantly enhanced, and the apoptosis rates were decreased, while the expressions of N-cadherin, vimentin and slug were up-regulated. However, the results are opposite via the over-expression of CACNA1C-AS2. Conclusion CACNA1C-AS2 enhances the proliferation, invasion and migration of esophageal cancer cells by promoting EMT.

Objective To study the regulation of D816V mutation of III tyrosine kinase receptor KIT on RNA binding proteins HNRNPL and HNRNPK. Methods In COS-1 cells, wild-type KIT or KIT D816V mutation were expressed alone or together with HNRNPL or HNRNPK. Activation of KIT and phosphorylation of HNRNPL and HNRNPK were detected by immunoprecipitation and Western blot analysis. The localization of KIT, HNRNPL and HNRNPK in COS-1 cells were examined by confocal microscopy. Results Wild-type KIT needs to bind its ligand stem cell factor (SCF) for phosphorylation, while KIT D816V could auto-phosphorylation without SCF stimulation. In addition, KIT D816V can induce phosphorylation of HNRNPL and HNRNPK, which is not possible in wild-type KIT. HNRNPL and HNRNPK are expressed in the nucleus, and wild-type KIT is expressed in cytosol and cell membrane, while KIT D816V is mainly found in cytosol. Conclusion Wild-type KIT needs SCF binding for activation, while KIT D816V can autoactivate without SCF stimulation, and induces phosphorylation of HNRNPL and HNRNPK specifically.

Objective To investigate the regulatory effect of wingless gene 7a (Wnt7a) on Bacille Calmette Guerin (BCG) induced autophagy in alveolar epithelial cells. Methods The alveolar epithelial cells of TC-1 mice were treated with interfering Wnt7a lentivirus and BCG alone or together in four groups, namely, small interfering RNA control (si-NC) group, si-NC combined with BCG group, Wnt7a small interfering RNA (si-Wnt7a) group, and si-Wnt7a combined with BCG group. The expressions of Wnt7a, microtubule associated protein 1 light chain 3 (LC3), P62, and autophagy related gene 5 (ATG5) were detected by Western blot analysis, and the distribution of LC3 was detected by immunofluorescence cytochemical staining. Results Compared with those in the si-NC group, Wnt7a, ATG5 and LC3 expressions increased and green fluorescent spots of LC3 significantly increased in the BCG infected TC-1 cells; the expressions of Wnt7a, LC3, P62, and ATG5 and green fluorescent spots of LC3 significantly decreased in the si-Wnt7a combined with BCG group comparing with those in the si-NC combined with BCG group. Conclusion Knocking down Wnt7a inhibits BCG induced autophagy in mouse alveolar epithelial cells.

Inflammatory bowel disease (IBD) is a chronic nonspecific inflammatory disease of the intestine, with unknown etiology and the incidence is increasing year by year. Traditional treatment has limited effect. Mesenchymal stem cell-derived exosomes (MSC-Exos) are a group of nano-sized extracellular vesicles. Their function is equivalent to that of mesenchymal stem cells (MSCs), with no tumorigenicity and high safety. They represent a novel cell-free therapy. It has been shown that MSC-Exos can improve IBD by effects including anti-inflammation, antioxidant stress, repairing intestinal mucosal barrier and immune regulation. However, their clinical application still faces some problems, such as the lack of standardized production technology, lack of specific IBD diagnostic molecules and anti-intestinal fibrosis.

Objective To investigate the effects of C-X-C motif chemokine ligand 1 (CXCL1) and its receptor CXCR2 on the cerebral endothelial cytoskeleton rearrangement and permeability in the inflammation of septic encephalopathy. Methods The murine model of septic encephalopathy was established by intraperitoneal injection of LPS (10 mg/kg). The levels of TNF-α and CXCL1 in the whole brain tissue were detected by ELISA. The expression of CXCR2 was detected by Western blot analysis after bEND.3 cells were stimulated with 500 ng/mL LPS and 200 ng/mL TNF-α. After treated with CXCL1(150 ng/mL), the changes of endothelial filamentous actin (F-actin) rearrangement in bEND.3 cells were observed by immuno-fluorescence staining. In the cerebral endothelial permeability test, bEND.3 cells were randomly divided into PBS control group, CXCL1 group, and CXCL1 combined with CXCR2 antagonist SB225002 group. Then endothelial transwell permeability assay kit was used to detect the endothelial permeability changes. After stimulated with CXCL1 in bEND.3 cells, Western blot analysis was used to detect the expression of protein kinase B (AKT) and phosphorylated-AKT (p-AKT). Results Intraperitoneal injection of LPS significantly increased the levels of TNF-α and CXCL1 in the whole brain. LPS and TNF-α both upregulated the expression of CXCR2 protein in bEND.3 cells. CXCL1 stimulation induced the endothelial cytoskeleton contraction, increased paracellular gap formation and elevated endothelial permeability in bEND.3 cells, which was inhibited by the pretreatment with SB225002(CXCR2 antagonist). Furthermore, CXCL1 stimulation also enhanced the phosphorylation of AKT in bEND.3 cells. Conclusion CXCL1 induces the cytoskeleton contraction and increased permeability through AKT phosphorylation in bEND.3 cells, which can be effectively inhibited by CXCR2 antagonist SB225002.

Objective To identify the effect of exosomes derived from bone marrow mesenchymal stem cells (BMSCs) loaded with annexin A2 (ANXA2) on the proliferation, migration, invasion of prostate cancer cells, and the transplanted tumor of prostate cancer in nude mice growth, as well as the role of macrophages in this process. Methods BMSCs from BALB/c nude mice were isolated and cultured. BMSCs were infected with lentiviral plasmids loaded with ANXA2. Exosomes were isolated and then added to treat THP-1 macrophages. ELISA was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), IL-6 and IL-10 in the cell supernatant culture fluid; After co-culturing the exosomes-treated macrophages and prostate cancer cells, CCK-8 assay was used to detect the cell proliferation activity. Also, Transwell^TM chamber were used to detect the cell invasion and migration. A nude mouse xenograft model of prostate cancer was constructed by injecting PC-3 human prostate cancer cells, the modeled nude mice were randomly divided into a control group and an experimental group, with 8 mice in each group. The nude mice in experimental group were injected with 1 mL of Exo-ANXA2 through the tail vein, while the control group was injected with the same amount of PBS, on days 0, 3, 6, 9, 12, 15, 18, and 21 after injection. Then the tumor volume was measured and calculated with vernier calipers. The nude mice were sacrificed at 21 days with their the tumor mass measured. The immunohistochemical staining method was used to detect the expression of antigen KI-67 (ki67) and CD163 expression in tumor tissue. Results The cells isolated from bone marrow showed high expression of CD90 and CD44 on the surface, and low expression of CD34 and CD45, and had strong osteogenic adipogenic differentiation ability, indicating that BMSCs were successfully obtained. After infection with the lentiviral plasmid carrying ANXA2, a strong green fluorescent protein expression in BMSCs, and Exo-ANXA2 was isolated. After Exo-ANXA2 treatment, the levels of TNF-α and IL-6 in THP-1 cells increased significantly, while the levels of IL-10 and IL-13 decreased significantly. Exo-ANXA2 treatment of macrophages significantly inhibited Exo-ANXA2 and promoted the proliferation, invasion and migration of PC-3 cells. After injecting Exo-ANXA2 into nude mice with prostate cancer cells transplantation, the tumor tissue volume of nude mice was significantly reduced on the 6th, 9th, 12th, 15th, 18th, and 21st days, and the tumor mass of nude mice was also significantly reduced on the 21st day. In addition, the positive expression rates of ki67 and CD163 in tumor tissues were significantly reduced. Conclusion Exo-ANXA2 can inhibit the proliferation, invasion and migration of prostate cancer cells, and suppress the growth of prostate cancer xenografts in nude mice by reducing M2 macrophages.

Objective To investigate the anti-DNA damage role of Sigma factor E (SigE) and its regulation mechanism of DNA damage repair in Mycobacterium smegmatis(MS). Methods The SigE gene of Mycobacterium smegmatis was cloned into plasmid pMV261 to construct recombinant plasmid pMV261(+)-SigE, and the inserted gene was verified by sequencing. The recombinant plasmid was electrically transformed into Mycobacterium smegmatis to construct SigE over-expression strain, and the expression of SigE was detected by Western blot analysis. The Mycobacterium smegmatis containing pMV261 plasmid was used as the control strain. Growth differences between the two stains were monitored by measuring the 600 nm absorbance (A₆₀₀) value of the bacterial culture suspension. The survival rate differences between two kinds of strains which were treated with three kinds of DNA damaging agents including ultraviolet ray (UV), cisplatin (DDP), and mitomycin C (MMC) were detected by colony forming unit (CFU) counting assay. DNA damage repair pathways of Mycobacteria were analyzed through bioinformatics and SigE-related genes were screened. The relative expression levels of these genes possibly related to the SigE against DNA damage were detected by real-time fluorescence quantitative PCR. Results SigE over-expression strain pMV261(+)-SigE/MS was constructed and the expression of SigE in Mycobacterium smegmatis was detected. Compared with the control strain, the SigE over-expression strain grew more slowly and entered the growth plateau later; survival rate analysis found that SigE over-expression strain was more resistant to three DNA damaging agents including UV, DDP, and MMC. Bioinformatic analysis indicated that SigE gene was closely related to DNA damage repair genes recA, single-strand DNA binding protein, (ssb), and dnaE2; the expression levels of recA, dnaE2, and ssb in SigE over-expression strain all increased with varying degrees compared with those in the control strain. Conclusion SigE plays an important role in inhibiting the DNA damage of Mycobacterium smegmatis, whose mechanism is closely related to the regulation of DNA damage repair.

Objective To investigate the therapeutic effect of bone marrow cell adoptive therapy on metabolic-dysfunction-associated fatty liver disease (MAFLD) in mice and its possible cell population. Methods The staining was used to detect the liver lesions of MAFLD in C57BL/6 mice induced by methionine and choline deficiency diet (MCD) and the adoptive therapeutic effect of bone marrow cells on MAFLD was evaluated by detecting the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The mRNA expressions of low density lipoprotein receptor (LDLR) and interleukin-4 (IL-4) in liver immune cells (including T, NKT, Kupffer cells and other cell populations) were detected by real-time quantitative PCR. The bone marrow cells labeled with 5, 6- carboxyfluorescein diacetate succinimidyl ester (CFSE) were injected into the tail vein of mice. The proportion of CFSE positive cells in liver tissue was observed by the frozen section, and the proportion of labeled cells in the liver and spleen was tracked by flow cytometry. The expression of CD3, CD4, CD8, NK1.1, CD11b and Gr-1 in CFSE labeled adoptive cells was detected by flow cytometry. The intracellular lipid content of NKT cells in liver tissue was evaluated by Nile Red lipid staining. Results The injury of liver tissue and the levels of serum ALT and AST in MAFLD mice were significantly reduced. At the same time, liver immune cells up-regulated the expression of IL-4 and LDLR. LDLR knockout mice induced more severe MAFLD after giving MCD diet. Bone marrow adoptive cells had a significant therapeutic effect and differentiated more NKT cells to colonize the liver. At the same time, the intracellular lipids of these NKT cells increased significantly. Conclusion Bone marrow cell adoptive therapy can reduce liver injury in MAFLD mice by differentiating more NKT cells and increasing the intracellular lipid content of these cells.