1.Decreased IgE antibody formation in mice treated with polyadenyic pollyuridylic acid and polyinosinic polycytidylic acid.
Bong Ki LEE ; Jeon Soo SHIN ; Min Kyung CHU ; Jung Koo YOUN
Journal of the Korean Society for Microbiology 1993;28(2):165-174
No abstract available.
Animals
;
Antibody Formation*
;
Immunoglobulin E*
;
Mice*
;
Poly I-C*
2.The Genotyping by the Nucleotide, Length, and Quantitative Polymorphism of C-stretch of HVR I of mtDNA D-Loop.
Ji Young KIM ; Kang Nam JIN ; Hyung Seok KIM ; Jong Tae PARK
Korean Journal of Legal Medicine 2008;32(1):72-85
To analyze the nucleotide polymorphism and length polymorphism of C-stretch of HVR 1 (Hypervariable Region I) of mtDNA D-loop in the maternal lineages in Koreans, sequencing of C-stretch, GeneScan analysis and cloning of a cases were performed in 266 random objects and 128 families which were confirmed by STR analysis of autosome, X chromosome and nucleotide polymorphism of mitochondrial DNA D-loop. C- stretch was classified into two groups. First group (the group of nucleotide polymorphism 74.2%) has 14 bases which show nucleotide polymorphism composed of adenine, cytosine and thymine without length polymorphism and second group (poly C tract 25.8%) shows length polymorphism by the changes of the number of adenine and cytosine. The patterns of nucleotide polymorphism were as follows: A4C5TC4 (64.8%), A4C5TC2TC (0.8%), A4C4TTC2TC (0.8%), A4C3TCTC4 (5.5%), A4C2TC2TC4 (0.8%), A4CTC3TC4 (0.8%), A4TC4TC4 (0.8%). The pattern of length polymorphism of poly C tract were as follows : LP10-16 (3.0%), LP11-16 (6.1%), LP11-17 (27.2%), LP11-18 (6.1%), LP12-16 (12.1%), LP12- 17 (39.4%), LP12-18 (3.0%), LP13-16 (3.0%) The copy number ratios of each fragment length in the same pattern of length polymorphism were various, and this data were valuable in individual identification because of the quantitative polymorphism of each fragment length. The correlation coefficients of copy number ratio of each fragment length in the families were more than 0.98 in 84.8% (28 of 33 families) of cases, which was interpreted as maternal inheritance of the copy number ratio of fragment length. However, in some cases (9.1%, 3 of 33 families), the correlation value of fragment length were less than 0.96 and in 2 cases of 33 families (6.1%), were 0.89 and 0.75 which suggested a possibility of mutation in the quantitative polymorphism although the length polymorphism were maternally inherited. From the above results, sequencing analysis of nucleotide polymorphism and GeneScan analysis of C- stretch must be combined to clearly identify the nucleotide polymorphism, length polymorphism, and quantitative polymorphism of C-stretch of HVR 1 of mtDNA D-loop.
Adenine
;
Clone Cells
;
Cloning, Organism
;
Coat Protein Complex I
;
Cytosine
;
DNA, Mitochondrial
;
Humans
;
Poly C
;
Thymine
;
Wills
;
X Chromosome
3.Expression profiles and function of Toll-like receptors in human corneal epithelia.
Xin-yi WU ; Jian-lu GAO ; Mei-yu REN
Chinese Medical Journal 2007;120(10):893-897
BACKGROUNDToll-like receptors play an important role in the human immune system. This study was conducted to investigate the expression profiles and function of Toll-like receptor (TLR) 1 - 9 in human corneal epithelium.
METHODSThe expression of TLR1 - 9 mRNA in 20 human donor corneal epithelia samples abraded during photorefractive keratotomy (PRK) and cultivated telomerase-immortalized human corneal epithelial cells (THCEs) was examined by semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis. Human peripheral blood mononuclear cells (PBMCs) were used as positive controls. The expression of the TLR2 and TLR4 proteins was detected by Western analysis. ELISA was used to detect IL-8 secretion from THCEs challenged with ligands for TLR3 and TLR4 with and without antibody blockade.
RESULTSThe expression of TLR1 - 9 at the mRNA level was detected in the epithelia of 20 patients and in THCE. Significant differences among individuals were observed. One patient was found to lack of the expression of TLR3, 4, 6 and 8, whereas another did not express TLR5. The expression of TLR2 and TLR4 protein was detected in human corneal epithelial cells. As THCE cells express TLR1 - 9, cells were challenged with lipopolysaccharides (LPS) and poly I:C to determine whether TLR4 and TLR3 were functional. The results showed that secretion of IL-8 by cells stimulated with LPS and Poly I:C was 7 to 10 fold greater than secretion by unchallenged cells. Blocking TLR4 with an anti-TLR4 antibody significantly inhibited the LPS-induced IL-8 production by THCE (P < 0.05).
CONCLUSIONHuman corneal epithelial cells express multiple TLRs and are able to recognize LPS and poly I:C. Different expression profiles among individuals suggest that differences in the susceptibilities and sensitivities to bacterial and viral infection in human populations relate to different patterns of TLR expression.
Blotting, Western ; Epithelium, Corneal ; metabolism ; Humans ; Lipopolysaccharides ; pharmacology ; Poly I-C ; pharmacology ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Toll-Like Receptors ; analysis ; genetics ; physiology
4.Induction of Indoleamine 2,3-dioxygenase by Pre-treatment with Poly(I:C) May Enhance the Efficacy of MSC Treatment in DSS-induced Colitis.
Da Bin RYU ; Ji Young LIM ; Sung Eun LEE ; Gyeongsin PARK ; Chang Ki MIN
Immune Network 2016;16(6):358-365
Mesenchymal stem cells (MSCs) have been used experimentally for treating inflammatory disorders, partly owing to their immunosuppressive properties. The goal of the study was to determine whether TLR ligands can enhance the therapeutic efficacy of bone marrow-derived MSCs for the treatment of inflammatory bowel disease. Mice (C57BL6) were administered with 4% dextran sulfate sodium (DSS) in drinking water for 7 days and injected with MSCs on days 1 and 3 following DSS ingestion. Our results demonstrated that among various TLR ligands, MSCs treated with polyinosinic-polycytidylic acid [poly(I:C)], which is a TLR3 ligand, more profoundly induced IDO, which is a therapeutically relevant immunosuppressive factor, without any observable phenotype change in vitro. The poly(I:C)-treated MSCs attenuated the pathologic severity of DSS-induced murine colitis when injected i.p. but not i.v. In summary, preconditioning MSCs with poly(I:C) might improve their efficacy in treating DSS-induced colitis, and this effect at least partly depends on the enhancement of their immunosuppressive activity through increasing their production of IDO.
Animals
;
Colitis*
;
Dextran Sulfate
;
Drinking Water
;
Eating
;
In Vitro Techniques
;
Indoleamine-Pyrrole 2,3,-Dioxygenase*
;
Inflammatory Bowel Diseases
;
Ligands
;
Mesenchymal Stromal Cells
;
Mice
;
Phenotype
;
Poly I-C
;
Toll-Like Receptors
6.Polyinosine-polycytidylic acid promotes excessive iodine intake induced thyroiditis in non-obese diabetic mice via Toll-like receptor 3 mediated inflammation.
Ya-nan SHI ; Feng-hua LIU ; Xiu-jie YU ; Ze-bing LIU ; Qing-xin LI ; Ji-hong YUAN ; Xiao-yi ZANG ; Lan-ying LI
Chinese Medical Journal 2013;126(4):703-710
BACKGROUNDExcessive iodine intake and viral infection are recognized as both critical factors associated with autoimmune thyroid diseases. Toll-like receptors (TLRs) have been reported to play an important role in autoimmune and inflammatory disorders. In this study, we aimed to clarify the possible mechanism of TLR3 involved in polyinosine-polycytidylic acid (poly(I:C)) promoting excessive iodine intake induced thyroiditis in non-obese diabetic (NOD) mice.
METHODSBoth NOD and BALB/c mice were randomly assigned to four groups: control group (n = 5), high iodine intake (HI) group (n = 7), poly(I:C) group (n = 7) and combination of excessive iodine and poly(I:C) injection (HIP) group (n = 7). After 8 weeks, mice were weighed and blood samples were collected. All the mice were sacrificed before dissection of spleen and thyroid gland. Then, thyroid histology, thyroid secreted hormone, expression of CD3(+) cells and TLR3 as well as inflammatory mRNA level were evaluated.
RESULTSBoth NOD and BALB/c mice from HI and HIP group represented goiter and increasing thyroid relative weight. Thyroid histology evidence indicated that only HIP group of NOD mice showed severe thyroiditis with lymphocytes infiltration in majority of thyroid tissue, severe damage of follicles and general fibrosis. Immunofluorescence staining results displayed a large number of CD3(+) cells in HIP NOD mice. Real-time polymerase chain reaction (PCR) results suggested interferon (IFN)-α increased over 30 folds and IFN-γ expression was doubled compared with control group, but interleukin (IL)-4 remained unchanged in HIP group of NOD mice thyroid. Meanwhile, over one third decrease of blood total thyroxine (TT4) and increased thyroid-stimulating hormone (TSH) was observed in HIP group of NOD mice. Only HIP group of NOD mice represented significantly elevation of TLR3 expression.
CONCLUSIONPoly(I:C) enhanced excessive dietary iodine induced thyroiditis in NOD mice through increasing TLR3 mediated inflammation.
Animals ; Female ; Inflammation ; chemically induced ; metabolism ; Iodine ; toxicity ; Mice ; Mice, Inbred NOD ; Poly I-C ; pharmacology ; Thyroiditis ; chemically induced ; immunology ; metabolism ; Toll-Like Receptor 3 ; metabolism
7.Viperin Differentially Induces Interferon-Stimulated Genes in Distinct Cell Types
Jeong Jin KIM ; Ku Sul KIM ; John EOM ; Jae Bong LEE ; Jun Young SEO
Immune Network 2019;19(5):e33-
Viperin is an IFN-stimulated gene (ISG)-encoded protein that was identified in human primary macrophages treated with IFN-γ and in human primary fibroblasts infected with cytomegalovirus (CMV). This protein plays multiple roles in various cell types. It inhibits viral replication, mediates signaling pathways, and regulates cellular metabolism. Recent studies have shown that viperin inhibits IFN expression in macrophages, while it enhances TLR7 and TLR9-mediated IFN production in plasmacytoid dendritic cells, suggesting that viperin can play different roles in activation of the same pathway in different cell types. Viperin also controls induction of ISGs in macrophages. However, the effect of viperin on induction of ISGs in cell types other than macrophages is unknown. Here, we show that viperin differentially induces ISGs in 2 distinct cell types, macrophages and fibroblasts isolated from wild type and viperin knockout mice. Unlike in bone marrow-derived macrophages (BMDMs), viperin downregulates the expression levels of ISGs such as bone marrow stromal cell antigen-2, Isg15, Isg54, myxovirus resistance dynamin like GTPase 2, and guanylate binding protein 2 in murine embryonic fibroblasts (MEFs) treated with type I or II IFN. However, viperin upregulates expression of these ISGs in both BMDMs and MEFs stimulated with polyinosinic-polycytidylic acid or CpG DNA and infected with murine CMV. The efficiency of viral entry is inversely proportional to the expression levels of ISGs in both cell types. The data indicate that viperin differentially regulates induction of ISGs in a cell type-dependent manner, which might provide different innate immune responses in distinct cell types against infections.
Animals
;
Carrier Proteins
;
Cytomegalovirus
;
Dendritic Cells
;
DNA
;
Dynamins
;
Fibroblasts
;
GTP Phosphohydrolases
;
Humans
;
Immunity, Innate
;
Interferons
;
Macrophages
;
Mesenchymal Stromal Cells
;
Metabolism
;
Mice
;
Mice, Knockout
;
Orthomyxoviridae
;
Poly I-C
8.Airway inflammation induced by Poly(I:C) stimulation in the late stage of respiratory syncytial virus infection in mice and its mechanism.
Ke-Ting ZHAO ; Xiao-Ru LONG ; Wei LI ; Jun XIE ; Luo REN ; Yu DENG ; Xiao-Hong XIE ; Na ZANG ; Li-Jia WANG ; En-Mei LIU
Chinese Journal of Contemporary Pediatrics 2016;18(5):455-459
OBJECTIVETo investigate the pathogenic mechanisms of airway inflammation and recurrent wheezing induced by recurrent respiratory virus infection after respiratory syncytial virus (RSV) infection.
METHODSSixty-four female BALB/c mice (aged 6-8 weeks) were randomly divided into four groups: control, RSV, Poly(I:C), and RSV+Poly(I:C) (n=16 each). The bronchoalveolar lavage fluid (BALF) was collected on the 3rd day after Poly(I:C) administration, and the total cell number and differential counts in BALF were determined. Hematoxylin-eosin staining was used to observe pulmonary pathological changes. The airway responsiveness was detected. ELISA was used to measure the levels of interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-13 (IL-13), matrix metallopeptidase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in BALF.
RESULTSCompared with the other three groups, the RSV+Poly(I:C) group had significant increases in the total number of inflammatory infiltrating cells in the airway, airway responsiveness, and MMP-9 level in BALF (P<0.05). The RSV+Poly(I:C) group showed more severe pulmonary tissue injuries compared with the control and RSV groups (P<0.01). Compared with the RSV group, the RSV+Poly(I:C) group showed significant reductions in the levels of IL-4 and TIMP-1 in BALF (P<0.01).
CONCLUSIONSViral re-infection in the late stage of RSV infection may cause an imbalance of MMP-9/TIMP-1 expression and thus contribute to aggravated airway inflammation.
Animals ; Asthma ; etiology ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; Lung ; pathology ; Matrix Metalloproteinase 9 ; analysis ; Mice ; Mice, Inbred BALB C ; Poly I-C ; pharmacology ; Respiratory Syncytial Virus Infections ; complications ; Tissue Inhibitor of Metalloproteinase-1 ; analysis
9.Effect of polyI: C on secretion of thymic stromal lymphopoietin and airway inflammation in mice with respiratory syncytial virus-induced asthma exacerbation.
Hu XIA ; Hua-peng YU ; Li-min LUO ; Shao-xi CAI
Journal of Southern Medical University 2011;31(3):434-437
OBJECTIVETo investigate the effects of polyinosinic-polycytidylic acid (polyI:C) on the production of thymic stromal lymphopoietin (TSLP) and airway inflammation in mice with exacerbated asthma induced by respiratory syncytial virus (RSV).
METHODSThirty-two female BALB/c mice were randomly divided into 4 groups, namely the PBS control group, OVA group, OVA/RSV group, and OVA/RSV/polyI:C group. In the latter 3 groups, the mice were sensitized by OVA and stimulated with nebulized OVA. RSV was inoculated into the nasal cavity of the sensitized mice and polyI:C (1 mg/kg) was intramuscularly administered. The airway response to metacholine was examined, and the serum levels of IL-4, IL-5, IL-13, and IFN-γ and TSLP in the supernatants of bronchoalveolar lavage fluid (BALF) were detected using ELISA. The total BALF cells, eosinophils, lymphocytes and neutrophils were counted. The lung specimens were collected to observe the inflammation with HE staining, and immunohistochemistry was employed to determine TSLP production in the airway epithelial cells.
RESULTSThe mice in RSV/OVA/polyI:C group showed a significantly lower airway responsiveness to metacholine than those in OVA/RSV group (P<0.01). Compared with OVA/RSV group, RSV/OVA/polyI:C group showed significantly lower serum levels of IL-4, IL-5, IL-13 and TSLP in BALF (P<0.05), with also lower total BALF cells, eosinophils and lymphocytes (P<0.05) and lessened infiltration of the airway inflammatory cells. Immunohistochemistry of TSLP also demonstrated a lower production of TSLP in the airway epithelial cells in RSV/OVA/polyI:C group than in OVA/RSV group.
CONCLUSIONSpolyI:C can inhibit the increase in TSLP production in the airway epithelial cells after RSV infection and relieve airway inflammation in mice with RSV-induced asthma exacerbation.
Animals ; Asthma ; blood ; metabolism ; virology ; Bronchoalveolar Lavage Fluid ; Cytokines ; secretion ; Female ; Inflammation ; pathology ; Interleukin-13 ; blood ; Interleukin-4 ; blood ; Interleukin-5 ; blood ; Mice ; Mice, Inbred BALB C ; Poly I-C ; pharmacology ; Respiratory Syncytial Virus Infections ; blood ; metabolism ; Respiratory Syncytial Viruses
10.Protective Effects of S-Adenosylmethionine and Its Combinations With Taurine and/or Betaine Against Lipopolysaccharide or Polyinosinic-polycytidylic Acid-induced Acute Hepatotoxicity.
Journal of Cancer Prevention 2016;21(3):152-163
BACKGROUND: Several mechanisms for the pathogenesis of many liver diseases are related with oxidative stress, endotoxins, and infections by many microorganisms. These can lead to chronic hepatitis, cirrhosis, and even liver cancer. The aim of this study was to evaluate the effects of S-adenosylmethionine (SAMe) and its combinations with taurine and/or betaine against hepatotoxicites induced by lipopolysaccharide (LPS) or polyinosinic-polycytidylic acid (polyI:C). METHODS: RAW 264.7 macrophage cells and seven-week-old male C57BL/6 mice were pretreated with SAMe (SAM or AdoMet), taurine, and/or betaine. In order to mimic hepatic injury like endotoxemia or viral infection, cells and mice were treated with LPS or polyI:C. Concentrations of glutathione (GSH), mRNA expressions of GSH synthesizing enzymes, and inflammatory markers were measured by biochemical assays and quantitative real-time PCR. RESULTS: In RAW 264.7 cells and mice, pretreatment of SAMe alone or SAMe with taurine and/or betaine attenuated the decrease in GSH levels and mRNA expressions of GSH synthesizing enzymes. In addition, pretreatment of SAMe with taurine and/or betaine prevented the excessive increase in inflammatory mediators produced by LPS or polyI:C treatment. CONCLUSIONS: Treatment with SAMe in combination with taurine and betaine, would have anti-oxidant functions in addition to anti-inflammatory action against bacterial and/or viral inflammation.
Animals
;
Betaine*
;
Endotoxemia
;
Endotoxins
;
Fibrosis
;
Glutathione
;
Hepatitis, Chronic
;
Humans
;
Inflammation
;
Lipopolysaccharides
;
Liver Diseases
;
Liver Neoplasms
;
Macrophages
;
Male
;
Mice
;
Oxidative Stress
;
Poly I-C
;
RAW 264.7 Cells
;
Real-Time Polymerase Chain Reaction
;
RNA, Messenger
;
S-Adenosylmethionine*
;
Taurine*