2.Intrinsic Cellular Defenses against Virus Infection by Antiviral Type I Interferon.
Kyung Hyun BOO ; Joo Sung YANG
Yonsei Medical Journal 2010;51(1):9-17
Intrinsic cellular defenses are non-specific antiviral activities by recognizing pathogen-associated molecular patterns (PAMPs). Toll-like receptors (TLRs), one of the pathogen recognize receptor (PRR), sense various microbial ligands. Especially, TLR2, TLR3, TLR4, TLR7, TLR8 and TLR9 recognize viral ligands such as glycoprotein, single- or double-stranded RNA and CpG nucleotides. The binding of viral ligands to TLRs transmits its signal to Toll/interleukin-1 receptor (TIR) to activate transcription factors via signal transduction pathway. Through activation of transcription factors, such as interferon regulatory factor-3, 5, and 7 (IRF-3, 5, 7) or nuclear factor-kappaB (NF-kappaB), type I interferons are induced, and antiviral proteins such as myxovirus-resistance protein (Mx) GTPase, RNA-dependent Protein Kinase (PKR), ribonuclease L (RNase L), Oligo-adenylate Synthetase (OAS) and Interferon Stimulated Gene (ISG) are further expressed. These antiviral proteins play an important role of antiviral resistancy against several viral pathogens in infected cells and further activate innate immune responses.
Animals
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GTP-Binding Proteins/metabolism
;
Humans
;
Interferon Regulatory Factors/metabolism
;
Interferon Type I/*metabolism/physiology
;
Models, Biological
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NF-kappa B/metabolism
;
Toll-Like Receptors/metabolism
;
Virus Diseases/*immunology/*metabolism/virology
;
eIF-2 Kinase/metabolism
3.Silencing of ICSBP/IRF8 expression in U226 cells and bone marrow mononuclear cells from patients with multiple myeloma.
Gu JIA ; Yan-Ping MA ; Ling ZHANG ; Yu-Jin LU ; Hong LU ; Ming-Xing CHANG ; Hua-Ping ZHANG
Journal of Experimental Hematology 2012;20(5):1127-1130
The objective of this study was to investigate expression of interferon regulatory factors (ICSBP/IRF8) and the potential role of DNA methylation in silencing ICSBP/IRF8 gene in multiple myeloma (MM) cell line U266 and bone marrow mononuclear cells from 10 MM patients (MM-BMMNC). The bone marrow mononuclear cells from 10 healthy persons (N-BMMNC) were collected and used as normal controls. Expression of ICSBP/IRF8 gene was detected by real-time fluorescence quantitative PCR (using 2(-ΔΔCT) to calculate); DNA methylation level of the ICSBP/IRF8 gene was measured using methylation-specific PCR (using the ratio of interest gene ICSBP/IRF8 and internal reference β-actin expression as results). The results showed that as compared with N-BMMNC the lower expression of ICSBP/IRF8 gene was found in U266 cells and MM-BMMNC, the hypermethylation of the CpG island in the ICSBP/IRF8 promoter was observed, there were significant differences between N-BMMNC and MM-BMMNC or U266 cells (P < 0.05). It is concluded that the expression of ICSBP/IRF8 gene can be silenced in the MM-BMMNC and U226 cells. As the hypermethylation of CpG island in ICSBP/IRF8 promoter is a frequent event in MM cells, the ICSBP/IRF8 gene silencing caused by DNA methylation may take part in the pathogenesis and development of MM.
Bone Marrow Cells
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metabolism
;
Case-Control Studies
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Cell Line, Tumor
;
DNA Methylation
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Gene Silencing
;
Humans
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Interferon Regulatory Factors
;
genetics
;
metabolism
;
Multiple Myeloma
;
genetics
;
metabolism
4.Study on mechanisms of the expression regulation of interferon-induced gene RIG-G.
Dong LI ; Shu XIAO ; Xiao-rong PAN ; Ye-jiang LOU ; Pei-min JIA ; Jian-hua TONG
Chinese Journal of Medical Genetics 2007;24(6):625-628
OBJECTIVETo investigate the molecular mechanisms of the expression regulation of retinoic acidinduced gene G (RIG-G) by interferon alpha (IFNalpha).
METHODSRIG-G promoter region was analyzed by bioinformatics. The functional activities of RIG-G promoter with or without IFNalpha were detected by luciferase reporter assay and electrophoretic mobility shift assay (EMSA).
RESULTSRIG-G promoter region contained two well-conserved IFN-stimulated response elements (ISREs). Both ISRE I and ISRE II showed their effective binding abilities with signal transducer and activator of transcription 1 (STAT1). In HT1080 cells, in contrast with the empty plasmid pXP2, pXP2-A reporter construct containing intact ISRE I and ISRE II showed a significant higher baseline expression (1741.2 +/- 517.5) which could be further enhanced up to three-four folds by IFNalpha (5338.7 +/- 1226.9, P < 0.05). However, the luciferase activity of pXP2-A as well as its IFNalpha inducibility could be abrogated in STAT1-deficient U3A cells (from 1741.2 +/- 517.5 to 406.1 +/- 103.2, P < 0.05), indicating that the STAT1 protein was a prerequisite for the activities of ISRE I and ISRE II.
CONCLUSIONISREs present in RIG-G promoter region are molecular basis of IFNalpha induced RIG-G expression. RIG-G is a target gene directly regulated by STAT1 protein and should play a key role in IFNalpha signaling pathways.
Base Sequence ; Cells, Cultured ; Gene Expression Regulation ; drug effects ; physiology ; Humans ; Interferon Regulatory Factor-1 ; genetics ; metabolism ; Interferon Regulatory Factors ; genetics ; metabolism ; Interferon-alpha ; pharmacology ; physiology ; Interferons ; physiology ; Molecular Sequence Data ; Promoter Regions, Genetic ; drug effects ; genetics ; physiology ; STAT1 Transcription Factor ; metabolism
5.Association between AOX1, IRF4 methylation in peripheral blood leukocyte DNA and the risks of breast cancer: a case-control study.
H ZHANG ; Y P LIU ; A Q GE ; X WANG ; H R SUN ; H R BI ; D PANG ; Y S ZHAO
Chinese Journal of Epidemiology 2018;39(9):1265-1269
Objective: To understand the relationship between AOX1, IRF4 gene methylation status in peripheral blood leukocyte DNA, as well as its interaction with environmental factors, and the risk of breast cancer. Methods: A case-control study was conducted among 401 breast cancer patients and 555 cancer-free controls selected from 2010 to 2014. Methylation sensitive-high resolution melting curve analysis was used to detect the methylation status of AOX1 and IRF4. The multiplication interaction effect between genes' methylation and environmental factors on the risk of breast cancer was analyzed by using unconditional logistic regression, and Excel software was used to analyze the additive interaction effect. Results: Individuals without AOX1 methylation had a 1.37-fold (95%CI: 1.02-1.84) higher breast cancer risk compared to individuals with AOX1 methylation. AOX1 methylation interacted with fungi intake (OR=2.06, 95%CI: 1.12-3.79) and physical activity (OR=2.18, 95%CI: 1.16-4.09) synergistically, on the risk for breast cancer, but no additive interaction effects were observed. Non-methylation of IRF4 could increase the risk for breast cancer, with statistical significance (OR=1.71, 95%CI: 0.99-7.43). Neither multiplication nor additive interactions were observed between IRF4 methylation and environmental factors. Conclusion: Non-methylation of AOX1 and IRF4 were a risk factors for breast cancer.
Aldehyde Oxidase/genetics*
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Breast Neoplasms/genetics*
;
Case-Control Studies
;
DNA Methylation/genetics*
;
Female
;
Genetic Predisposition to Disease
;
Humans
;
Interferon Regulatory Factors/genetics*
;
Leukocytes/metabolism*
6.Mechanism of transforming growth factor- β1 induce renal fibrosis based on transcriptome sequencing analysis.
Huanan LI ; Peifen LI ; Shanyi LI ; Xueying ZHANG ; Xinru DONG ; Ming YANG ; Weigan SHEN
Journal of Zhejiang University. Medical sciences 2023;52(5):594-604
OBJECTIVES:
To explore the mechanism of transforming growth factor-β1 (TGF-β1) induce renal fibrosis.
METHODS:
Renal fibroblast NRK-49F cells treated with and without TGF-β1 were subjected to RNA-seq analysis. DESeq2 was used for analysis. Differentially expressed genes were screened with the criteria of false discovery rate<0.05 and l o g 2 F C >1. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed for differentially expressed genes. Genes encoding transcription factors were further screened for differential expression genes. Then, the expression of these genes during renal fibrosis was verified using unilateral ureteral obstruction (UUO)-induced mouse renal fibrosis model and a public gene expression dataset (GSE104954).
RESULTS:
After TGF-β1 treatment for 6, 12 and 24 h, 552, 1209 and 1028 differentially expressed genes were identified, respectively. GO analysis indicated that these genes were significantly enriched in development, cell death, and cell migration. KEGG pathway analysis showed that in the early stage of TGF-β1 induction (TGF-β1 treatment for 6 h), the changes in Hippo, TGF-β and Wnt signaling pathways were observed, while in the late stage of TGF-β1 induction (TGF-β1 treatment for 24 h), the changes of extracellular matrix-receptor interaction, focal adhesion and adherens junction were mainly enriched. Among the 291 up-regulated differentially expressed genes treated with TGF-β1 for 6 h, 13 genes (Snai1, Irf8, Bhlhe40, Junb, Arid5a, Vdr, Lef1, Ahr, Foxo1, Myc, Tcf7, Foxc2, Glis1) encoded transcription factors. Validation in a cell model showed that TGF-β1 induced expression of 9 transcription factors (encoded by Snai1, Irf8, Bhlhe40, Junb, Arid5a, Vdr, Lef1, Myc, Tcf7), while the expression levels of the other 4 genes did not significantly change after TGF-β1 treatment. Validation results in UUO-induced mouse renal fibrosis model showed that Snai1, Irf8, Bhlhe40, Junb, Arid5a, Myc and Tcf7 were up-regulated after UUO, Vdr was down-regulated and there was no significant change in Lef1. Validation based on the GSE104954 dataset showed that IRF8 was significantly overexpressed in the renal tubulointerstitium of patients with diabetic nephropathy or IgA nephropathy, MYC was highly expressed in diabetic nephropathy, and the expressions of the other 7 genes were not significantly different compared with the control group.
CONCLUSIONS
TGF-β1 induces differentially expressed genes in renal fibroblasts, among which Irf8 and Myc were identified as potential targets of chronic kidney disease and renal fibrosis.
Mice
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Animals
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Humans
;
Transforming Growth Factor beta1/metabolism*
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Diabetic Nephropathies/pathology*
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Transcriptome
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Signal Transduction
;
Kidney
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Ureteral Obstruction/pathology*
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Fibrosis
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Interferon Regulatory Factors
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Transforming Growth Factor beta/metabolism*
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DNA-Binding Proteins/metabolism*
;
Transcription Factors/metabolism*
7.Effects of ozone exposure on percentage of CD4(+)CD25(high)Foxp(3+) regulatory T cells and mRNA expression of Foxp3 in asthmatic rats.
Ming WEI ; Ling TU ; Ying-hong LIANG ; Jia LIU ; Yan-jie GONG ; Jun-hua ZHANG ; Yi-hua ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(9):693-696
OBJECTIVETo investigate the effects of low-concentration ozone exposure on the percentage of CD4(+)CD25(high)Foxp(3+) regulatory T cells and the mRNA expression of transcription factor Foxp3 in asthmatic rats.
METHODSSixty male Wistar rats were randomly divided into 4 groups (n = 15 for each): normal control group, ovalbumin (OVA) exposure group, ozone exposure group, and OVA+ozone exposure group. The OVA exposure group was sensitized and challenged with OVA to establish an asthma model; the normal control group inhaled aerosolized saline; the ozone exposure group inhaled low-concentration ozone; the OVA+ozone exposure group inhaled low-concentration ozone before being challenged with aerosolized OVA every day. The percentage of CD4(+)CD25(high)Foxp(3+) regulatory T cells in CD4(+) T cells was determined by flow cytometry. The levels of interferon-γ (INF-γ) and interleukin 4 (IL-4) in peripheral blood and lung tissue were measured by enzyme-linked immunosorbent assay. The mRNA expression of Foxp3 in lung tissue was measured by PCR.
RESULTSThe percentages of CD4(+)CD25(high)Foxp(3+) regulatory T cells in OVA exposure group (6.12±1.03%) and ozone exposure group (5.87±1.26%) were significantly lower than that in normal control group (9.85±1.34%), and the percentage of CD4(+)CD25(high)Foxp(3+) regulatory T cells in OVA+ozone exposure group (3.31±0.85%) was significantly lower than those in normal control group and OVA exposure group (P < 0.01). The levels of IL-4 in plasma and lung tissue in OVA exposure group (plasma: 21.83±5.12 ng/L; lung tissue: 0.89±0.13 ng/L) were significantly higher than those in normal control group (plasma: 10.58±2.73 ng/L; lung tissue: 0.32±0.11 ng/L) (P < 0.01). The levels of IL-4 in plasma and lung tissue in OVA+ozone exposure group (plasma: 35.47±7.24 ng/L; lung tissue: 1.50±0.42 ng/L) were significantly higher than those in normal control group and OVA exposure group (P < 0.01). The levels of INF-γ in plasma and lung tissue in OVA exposure group (plasma: 61.78±23.45 ng/L; lung tissue: 0.69±0.21 ng/L] were significantly lower than those in normal control group [plasma: 158.89±60.23 ng/L; lung tissue: 1.86±0.29) (P < 0.01). The levels of INF-γ in plasma and lung tissue in OVA+ozone exposure group (plasma: 10.28±2.63 ng/L; lung tissue: 0.41±0.12 ng/L) were significantly lower than those in normal control group and OVA exposure group (P < 0.01). The mRNA expression of Foxp3 was significantly lower in the OVA+ ozone exposure group than in the normal control group (P < 0.05).
CONCLUSIONLow-concentration ozone exposure may decrease the number of CD4(+)CD25(high)Foxp(3+) regulatory T cells and inhibit the mRNA expression of Foxp3 to promote Th1/Th2 imbalance in asthmatic rats, suggesting that ozone exposure may be one of factors that induce asthma attack.
Animals ; Asthma ; metabolism ; Environmental Exposure ; Flow Cytometry ; Forkhead Transcription Factors ; genetics ; metabolism ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Lung ; metabolism ; pathology ; Male ; Ozone ; adverse effects ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; T-Lymphocytes, Regulatory ; drug effects ; metabolism ; Th1-Th2 Balance
8.Right leg mass with thrombocytopenia.
Wen-xiu YANG ; Li BAO ; Qi-lan LIU ; Shang-yi LIU
Chinese Journal of Pathology 2011;40(3):191-192
Antigens, CD20
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metabolism
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Female
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Follow-Up Studies
;
Humans
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Interferon Regulatory Factors
;
metabolism
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Leg
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pathology
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Lymphoma, Large B-Cell, Diffuse
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complications
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metabolism
;
pathology
;
surgery
;
Middle Aged
;
Proto-Oncogene Proteins c-bcl-2
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metabolism
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Skin Neoplasms
;
complications
;
metabolism
;
pathology
;
surgery
;
Thrombocytopenia
;
complications
9.Epstein-Barr virus positive anaplastic-like plasmacytoma: report of a case.
Wen-sheng LI ; Xiao-ge ZHOU ; Jian-lan XIE
Chinese Journal of Pathology 2013;42(3):203-204
ADP-ribosyl Cyclase 1
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metabolism
;
Aged
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Epstein-Barr Virus Infections
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Herpesvirus 4, Human
;
isolation & purification
;
Humans
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Interferon Regulatory Factors
;
metabolism
;
Ki-67 Antigen
;
metabolism
;
Male
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Nasal Cavity
;
Nose Neoplasms
;
metabolism
;
pathology
;
surgery
;
virology
;
Plasmacytoma
;
metabolism
;
pathology
;
surgery
;
virology
10.Development of Monoclonal Antibodies Against Human IRF-5 and Their Use in Identifying the Binding of IRF-5 to Nuclear Import Proteins Karyopherin-alpha1 and -beta1.
Soo In YEON ; Ju Ho YOUN ; Mi Hwa LIM ; Hye Ja LEE ; Young Mok KIM ; Ji Eun CHOI ; Jae Myun LEE ; Jeon Soo SHIN
Yonsei Medical Journal 2008;49(6):1023-1031
PURPOSE: IRF-5 is a direct transducer of virus-mediated and TLR-mediated signaling pathways for the expression of cytokines and chemokines which form homodimers or heterodimers with IRF-7. However, direct IRF-5-specific monoclonal antibodies (mAbs) are not available at present. These could be used to further evaluate the functions of IRF-5. In this study, we produced and characterized three mouse mAbs to human IRF-5. The binding of IRF-5 to nuclear import proteins was first identified using a mAb. MATERIALS AND METHODS: His-tagged human IRF-5 protein spanning amino acid residues 193- 257 was used as an antigen and three mAbs were produced. The mAbs were tested with ELISA, Western blot analysis (WB), immunofluorescent staining (IF), and immunoprecipitation (IP). In addition, the nuclear import protein which carried phosphorylated IRF-5 was identified using one of these mAbs. RESULTS: MAbs 5IRF8, 5IRF10 and 5IRF24 which reacted with the recombinant His-IRF-5(193-257) protein were produced. All mAbs bound to human IRF-5, but not to IRF-3 or IRF-7. They could be used for WB, IF, and IP studies. The binding of phosphorylated IRF-5 to karyopherin-alpha1 and -beta1 was also identified. CONCLUSION: Human IRF-5-specific mAbs are produced for studying the immunologic roles related to IRF-5. Phosphorylated IRF-5 is transported to the nucleus by binding to nuclear import proteins karyopherin-alpha1 and -beta1.
Animals
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*Antibodies, Monoclonal
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Base Sequence
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Cell Line
;
Cross Reactions
;
DNA Primers/genetics
;
Humans
;
Interferon Regulatory Factors/genetics/*immunology/*metabolism
;
Mice
;
Mice, Inbred BALB C
;
NIH 3T3 Cells
;
Protein Binding
;
Recombinant Proteins/genetics/immunology/metabolism
;
alpha Karyopherins/*metabolism
;
beta Karyopherins/*metabolism