Changes in lipid metabolism have been implicated in protection against infectious diseases. In the first experiment of this study, we measured clinical lipid parameters in a murine model where the unmethylated cytidin...Changes in lipid metabolism have been implicated in protection against infectious diseases. In the first experiment of this study, we measured clinical lipid parameters in a murine model where the unmethylated cytidine phosphate guanosine (CpG) oligodinucleotide (ODN1826), a Toll-like receptor 9 (TLR9) agonist was administered in combination with D-galactosamine (GalN) that caused relatively liver-specific inflammation and toxicity. In the control mice group injected with phosphate-buffered saline (PBS) (acute psychological stress model associated with blood sampling), the serum triglyceride (TG) levels showed a rapid decrease followed by a rebound at 24 h as we have recently reported. However, such a TG rebound was impaired in the CpG/GalN- and solely CpG-treated groups of mice despite an absence of liver injury based on serum alanine aminotransferase levels in the latter group. Thus, the stress-associated serum TG rebound was abrogated by the injection of a sub-hepatotoxic CpG dose. In the second experiment, we simply measured the hepatic CD36 and SACRB1 (the gene for scavenger receptor B1 (SR-B1)) transcripts after the i.p. administration of PBS, CpG or CpG/GalN. There was a remarkable elevation of hepatic CD36 transcript expression in both the CpG- and CpG/GalN-treated mice at 8 h post-CpG injection whereas the increase in the PBS-treated mice was slower than the former two groups, suggesting that hepatic CD36 transcript expression is more pronounced in the combined stress models than under psychological stress alone. The individual mice data showed that the increase in CD36 expression was accompanied by a reduction in SCARB1 mRNA, showing reciprocal regulation between these two genes. Together with our previously reported findings, these data suggest that, in a murine model combining psychological stress with TLR-triggered hepatic inflammation, the psychological stress facilitates liver uptake of plasma TG (and its components fatty acids), but the subsequent re-esterification and/or release of TG-rich lipoproteins from the liver is impaired due to the concomitant TLR-signaling. We hypothesize that lipid metabolism during acute stress shifts toward an elevated hepatic uptake of lipids due to concomitant TLR signaling, facilitating the clearance of bacterial lipids by the liver.展开更多
Toll-like receptors (TLRs) recognize specific motifs which are frequently present in bacteria, fungi, prokaryotes and viruses. Amongst TLRs, TLR9 can be activated by such bacterial or viral DNA fragments, immunoglobul...Toll-like receptors (TLRs) recognize specific motifs which are frequently present in bacteria, fungi, prokaryotes and viruses. Amongst TLRs, TLR9 can be activated by such bacterial or viral DNA fragments, immunoglobulin-DNA complexes or synthetic oligonucleotides, which all contain unmethylated cytosineguanine nucleotide sequences (CpGs). Emerging data indicate that TLR9 signaling has a role in, and may influence, colorectal carcinogenesis and colonic inflammation. CpGs are classified into three groups according to their influence on both the antigen-specific humoraland cellular immunity, and the production of type 1 interferons and proinflammatory cytokines. TLR9 activation via CpGs may serve as a new therapeutic target for several cancerous and various inflammatory conditions. Due to its probable anti-cancer effects, the application possibilities of TLR9-signaling modulation may be extremely diverse even in colorectal tumors. In this review we aimed to summarize the current knowledge about TLR-signaling in the pathogenesis and therapy of inflammatory bowel diseases and colorectal cancer. Due to the species-specific differences in TLR9 expression, however, one must be careful in translating the animal model data into the human system, because of the differences between CpG-oligodeoxynucleotide-responsive cells. TLR9 agonist DNA-based immunomodulatory sequences could also represent a promising therapeutic alternative in systemic inflammatory conditions and chronic colonic inflammations as their side effects are not significant.展开更多
BACKGROUND Toll-like receptors(TLRs)are the first line of host defense,and are involved in Helicobacter pylori(H.pylori)recognition and activation of both inflammatory and carcinogenic processes.The presence of single...BACKGROUND Toll-like receptors(TLRs)are the first line of host defense,and are involved in Helicobacter pylori(H.pylori)recognition and activation of both inflammatory and carcinogenic processes.The presence of single nucleotide polymorphisms(SNPs)in genes that activate the immune response may modulate the risk of precancerous lesions and gastric cancer(GC).Among them,Toll-like receptor 9(TLR9)polymorphisms have emerged with a risk factor of infectious diseases and cancer,however the studies are still inconclusive.AIM To evaluate whether TLR9 rs5743836 and rs187084 SNPs contribute to the risk of gastric carcinogenesis,and its influence on mRNA expression.METHODS A case-control study was conducted to evaluate two TLR9 SNPs(TLR9-1237 TCrs5743836 and TLR9-1486 CT-rs187084)in chronic gastritis(CG)and GC patients.A total of 609 DNA samples of peripheral blood[248 CG,161 GC,and 200 samples from healthy individuals(C)]were genotyped by polymerase chain reaction-restriction fragment length polymorphism.All samples were tested for the H.pylori infection using Hpx1 and Hpx2 primers.Quantitative polymerase chain reaction by TaqMan?assay was used to quantify TLR9 mRNA from fresh gastric tissues(48 GC,26 CG,and 14 C).RESULTS For TLR9-1237,the TC+CC or CC genotypes were associated with a higher risk of GC than C[recessive model odds ratio(OR)=5.01,95%confidence interval(CI):2.52-9.94,P<0.0001],and the CG(recessive model OR=4.63;95%CI:2.44-8.79,P<0.0001)groups.For TLR9-1486,an association between the CT+TT genotypes and increased risk of both GC(dominant model OR=2.72,95%CI:1.57-4.72,P<0.0001)and CG(dominant model OR=1.79,95%CI:1.15-2.79,P=0.0094)was observed when compared to the C group.Moreover,the presence of TLR9-1237 TC/CC+TLR9-1486 CC genotypes potentiate the risk for this neoplasm(OR=18.57;95%CI:5.06-68.15,P<0.0001).The TLR9 mRNA level was significantly higher in the GC group(RQ=9.24,P<0.0001)in relation to the CG group(RQ=1.55,P=0.0010)and normal mucosa(RQ=1.0).When the samples were grouped according to the polymorphic genotypes and the presence of H.pylori infection,an influence of TLR9-1237 TC+CC polymorphic genotypes(P=0.0083)and H.pylori infection(P<0.0001)was observed on the upregulation of mRNA expression.CONCLUSION Our findings show that TLR9 rs5743836 and rs187084 polymorphisms are associated with a higher risk of carcinogenesis gastric,and that TLR9 mRNA levels can be modulated by TLR9-1237 TC+CC variant genotypes and H.pylori infection.展开更多
The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. Th...The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. The study group included 90 patients （60 with chronic hepatitis B, and 30 with chronic hepatitis C）, and 20 healthy people served as control group. The protein and mRNA levels of TLR9 were detected by using flow cytometry and real-time PCR. The serum viral copies of HBV and HCV were measured in all patients, and the correlation between HBV-DNA copies or HCV-RNA copies and the TLR9 expression was analyzed. Our results demonstrated that HBV or HCV infection led to a decreased expression of TLR9 mRNA and protein compared to the control group （P〈0.05）. The TLR9 protein and mRNA levels were negatively correlated with serum viral copies of HBV and HCV （r=-0.632, r=-0.909, P〈0.01）. It was concluded that TLR9 mRNA and protein are down-regulated in PBMC of HBV-infected or HCV-infected patients, and they are negatively correlated with serum viral copies and play an important role in detecting viral replication of HBV and HCV.展开更多
Abnormal innate immune responses toward luminal bacteria play an important role in the pathogenesis of inflammatory bowel disease.It has been demonstrated that bacteria having CpG DNA ameliorate experimental colitis i...Abnormal innate immune responses toward luminal bacteria play an important role in the pathogenesis of inflammatory bowel disease.It has been demonstrated that bacteria having CpG DNA ameliorate experimental colitis in mice,and Toll-like receptor 9 (TLR9) signaling mediates the anti-inflammatory effects in mouse colonic inflammation.A gene variation in NOD2/CARD15 has been reported in Crohn's disease (CD) patients in Western countries,but this variation has not been identified in Japanese CD patients.Therefore,we hypothesized that TLR9 is a key factor in the development of ulcerative colitis (UC),and we investigated gene mutations and polymorphisms of TLR9 in Japanese UC patients.Three single nucleotide polymorphisms (SNPs) in TLR9 were identified in healthy controls,and were assessed in 48 UC patients and 47 healthy controls.Control subjects were matched for age,sex and date of blood sampling from among a subgroup of participants.We found that TLR9-1486CC,1174GG and 2848AA increase the risk of UC [odds ratio (OR) 2.64,95% confidence interval (95% CI):1.73-6.53,P=0.042],and TLR9-1486TT,1174AA and 2848GG decrease the risk of UC (OR 0.30,95% CI:0.10-0.94,P=0.039),although there were no correlations between SNPs and disease phenotype or TLR9 mRNA expression.These findings suggest that TLR9 polymorphisms are associated with increased susceptibility to UC.展开更多
目的探讨固有免疫信号通路分子IRAK-M在TLR9信号诱导肝组织淋巴细胞浸润过程中的作用。方法应用野生型B6小鼠及IRAK-M基因敲除小鼠,腹腔注射TLR9配体CpG寡核苷酸激活肝内TLR9信号系统,提取小鼠肝内浸润淋巴细胞,并采用流式细胞术检测肝...目的探讨固有免疫信号通路分子IRAK-M在TLR9信号诱导肝组织淋巴细胞浸润过程中的作用。方法应用野生型B6小鼠及IRAK-M基因敲除小鼠,腹腔注射TLR9配体CpG寡核苷酸激活肝内TLR9信号系统,提取小鼠肝内浸润淋巴细胞,并采用流式细胞术检测肝内浸润不同淋巴细胞群的比率及细胞因子的表达情况。结果 CpG ODN激活TLR9信号通路后,IRAK-M基因敲除小鼠肝内浸润CD4+和CD8+T细胞比率较野生型B6小鼠显著增加[CD4+:(22.50±0.73)% vs. (20.03±0.75)%;CD8+:(17.83±0.67)%vs.(15.55±0.75)%;P均<0.05];肝内淋巴细胞细胞因子白细胞介素-6(IL-6)和肿瘤坏死因子(TNF-α)合成增加[IL-6:(1.91±0.26)% vs. (0.93±0.12)%;TNF-α:(13.23±1.23)% vs. (8.16±0.66)%;P均<0.01]。结论 IRAK-M具有负调节肝内TLR9信号系统的作用。展开更多
目的：探讨Toll样受体（Tol- like receptors/TLRs）9基因rs187084位点多态性与子宫内膜异位症（EMs）发病风险的关系。方法采用DNA直接测序法检测104例EMs患者（EMs组）和100例非EMs患者（对照组）的TLR9基因rs187084位点的多态性，比...目的：探讨Toll样受体（Tol- like receptors/TLRs）9基因rs187084位点多态性与子宫内膜异位症（EMs）发病风险的关系。方法采用DNA直接测序法检测104例EMs患者（EMs组）和100例非EMs患者（对照组）的TLR9基因rs187084位点的多态性，比较各基因型和等位基因的分布情况。结果 EMs组和对照组中，TLR9基因rs187084位点的AA、AG、GG3种基因型频率分别为42.30%、28.85%、28.85%和52.00%、34.00%、14.00%，两组比较差异有统计学意义（P=0.036）；A、G等位基因频率分别为56.73%、43.27%和69.00%、31.00%，两组比较差异有统计学意义（P=0.01）；携带GG基因型明显增加EMs的发病风险（OR=1.911，95％CI 1.113～3.282）。结论 TLR9基因rs187084位点多态性与EMs的发病风险相关。展开更多
目的通过观察早产儿不同胎龄Toll样受体9(TLR9)的表达,探讨早产儿免疫功能低下的机制。方法采集2010年7月至2014年6月在上海市嘉定区妇幼保健院产科出生的活产新生儿的脐血229份,按胎龄分为4组,28~31周组,31~34周组,34~37周组,≥37周组...目的通过观察早产儿不同胎龄Toll样受体9(TLR9)的表达,探讨早产儿免疫功能低下的机制。方法采集2010年7月至2014年6月在上海市嘉定区妇幼保健院产科出生的活产新生儿的脐血229份,按胎龄分为4组,28~31周组,31~34周组,34~37周组,≥37周组,采用流式细胞术和实时荧光定量PCR方法,分别检测其TLR9的蛋白和m RNA表达情况,了解其与胎龄之间的关系,并分析m RNA和蛋白表达间的相关性。结果 TLR9阳性细胞率在28~31周组,31~34周组,34~37周组,≥37周组分别为(15.93±6.23)%,(11.63±6.70)%,(13.66±6.88)%,(20.51±12.06)%;其在胎龄28~31周较高,至31~34周逐渐下降至最低,两组差异有统计学意义(P<0.05);34~37周后TLR9阳性细胞率表达逐渐升高,至≥37周达最高,两胎龄组比较,差异具有统计学意义(P<0.05)。31~37周间新生儿脐血TLR9阳性细胞率与胎龄呈正相关(r=0.273,P=0.006)。TLR9 m RNA表达在28~31周组,31~34周组,34~37周组,≥37周组分别为(4.95±3.44)%,(8.89±8.49)%,(13.91±10.92)%,(7.19±7.11)%;其在28~36周逐渐升高,与胎龄呈正相关(r=0.355,P<0.001)。≥37周TLR9 m RNA表达量下降,该值虽高于28~31周,但差异无统计学意义(P>0.05)。相关性分析表明,同胎龄时期同样本新生儿的TLR9 m RNA和TLR9阳性细胞率之间存在负相关(r=-0.227,P=0.011)。结论 TLR9阳性细胞率和TLR9 m RNA表达在不同胎龄组新生儿间有差异,TLR9阳性细胞率表达在31~37周间随着胎龄的增加而增加,TLR9 m RNA在28~36周间随着胎龄的增加而增加。展开更多
AIM:To determine the expression of toll-like receptor 9(TLR9) in pancreatic tumor and the effects of cytosine phosphate-guanosine oligodeoxynucleotides 2216(CPG ODN2216) on biological behavior of pancreatic carcinoma ...AIM:To determine the expression of toll-like receptor 9(TLR9) in pancreatic tumor and the effects of cytosine phosphate-guanosine oligodeoxynucleotides 2216(CPG ODN2216) on biological behavior of pancreatic carcinoma cell line PANC-1 and explore their clinical significance.METHODS:The immunohistochemistry and Western blot were used to determine the expression of TLR9 protein in pancreatic cancer tissues,and immunofluorescence staining was performed to detect the TLR9 protein expression in pancreatic carcinoma cell line PANC-1.To assess the effects of CPG ODN2216 on the invasive property of Panc-1 cells,in vitro cell adhesion,wound-healing scrape,and invasion and cell colony formation were evaluated.RESULTS:TLR9 was highly expressed in pancreaticcancer tissues and PANC-1 cells.The percentage of positive cells expressing TLR9 protein in human pancreatic tissues,paracancerous tissues and normal tissues were 73.3%,33.3% and 20.0%,respectively,and the protein expression level of TLR9 was gradually descending(P < 0.05).In vitro tests in wound-healing scrape,cell adhesion,colony formation and matrigel invasion showed that the adhesion and motility of PANC-1 cells in CPG ODN 2216 treatment group were signif icantly lower than in the control group(P < 0.05).The cell growth assay showed that the proliferative ability of PANC-1 cells in treatment group was significantly decreased and CPG ODN2216 had an inhibitive effect in the growth of Panc-1 cells in a dose and time-dependent manner(P < 0.05).CONCLUSION:The gene of TLR9 is correlated with the invasive and metastatic potential of human pancreatic carcinoma,and CPG ODN2216 induces the inhibition of migration and invasion of Panc-1 cells.展开更多
Inflammation and infection play an important role in the pathogenesis of many cancers.Toll-like receptors（TLRs） are a class of pattern recognition receptors that recognize conserved components of microbes and trigge...Inflammation and infection play an important role in the pathogenesis of many cancers.Toll-like receptors（TLRs） are a class of pattern recognition receptors that recognize conserved components of microbes and trigger the immune response against invading microorganisms.Toll-like receptor 9（TLR9） recognizes non-methylated cytosine-phosphateguanosine（CpG） DNA sequences which are the surrogate for viral DNA.TLR9 may react to tumor development and progression during chronic inflammation that involves the tumor microenvironment.In order to study the role of TLR9 in cervical cancer,we analyzed the TLR9 expression in different types of HPV infection cervical cancer cells.Then we detected if CpG sequences influenced the TLR9 expression and the sensitivity to cisplatin（DDP） of these cervical cancer cells in vitro.The expression of TLR9 mRNA and protein in SiHa,Hela and C33A cells was detected by RT-PCR and Western blotting.Real-time PCR was used to examine the TLR9 expression changes induced by CpG.Chemosensitivity of the cervical cancer cells to cisplatin（DDP） was measured by MTT.It was observed that the expression of TLR9 mRNA and protein was increased gradually in SiHa（HPV16＋）,Hela（HPV18＋） and C33A（HPV-） cells.Low doses of CpG increased the TLR9 expression only in C33A（HPV-） cells,but not in SiHa（HPV16＋） and Hela（HPV18＋） cells.Furthermore,low dose of CpG significantly increased the sensitivity of C33A（HPV-） cells,but not that of SiHa（HPV16＋） and Hela（HPV18＋） cells.These results indicated that TLR9 may serve as a protective agent in HPV negative cervical cancer cells.It was concluded that TLR9 could improve the sensitivity to DDP in HPV negative cervical cancer cells and might represent a potential therapeutic option in clinical practice.展开更多
Klebsiella has been considered as initiator of AS （ankylosing spondylitis） for nearly four decades. This study aimed to demonstrate that Klebsiella triggers ERS （endoplasmic reticulum stress） and HLA-B27 heavy cha...Klebsiella has been considered as initiator of AS （ankylosing spondylitis） for nearly four decades. This study aimed to demonstrate that Klebsiella triggers ERS （endoplasmic reticulum stress） and HLA-B27 heavy chain misfolding. CA46 cells or splenocytes obtained from wild-type, MyD88/ or TLR9/ mice were stimulated with KP （Klebsiella pneumoniae） or its components including CPS （capsule polysaccharide）, LPS （lipopolysaccharide）, and KP gDNA （genomic deoxyribonucleic acid） respectively for 24 h and 48 h. The activation of ERS-related signaling was detected by Western blotting or RT-PCR, and the level of misfolded HLA-B27 was determined by non-reducing protein gel electrophoresis and Western blotting. The protein expression of BiP/Grp78 and calreticulin, the alternative splicing of XBP-1 mRNA （messenger ribonucleic acid）, and the activation of caspase-12 and p38 were increased in a dose-dependent manner in HLA-B27-expressing CA46 cells after treatment with decapsulated KP. We also demonstrate that the EP, S-inducing effects occur via the TLR （Toll-like receptor）/MyD88-dependent signaling pathway. Significantly, HLA-B27 misfolding was also detected in decapsulated KP-treated B27-expressing cells. These results suggest that the non-antigen-specific induction of ERS and B27 misfoiding through TLR/MyD88 signaling might promote KP antigen-initiated autoreactive responses via the presentation of misfolded B27, and that small-molecules targeting TLRs might have potential as novel therapeutic agents for AS.展开更多
文摘Changes in lipid metabolism have been implicated in protection against infectious diseases. In the first experiment of this study, we measured clinical lipid parameters in a murine model where the unmethylated cytidine phosphate guanosine (CpG) oligodinucleotide (ODN1826), a Toll-like receptor 9 (TLR9) agonist was administered in combination with D-galactosamine (GalN) that caused relatively liver-specific inflammation and toxicity. In the control mice group injected with phosphate-buffered saline (PBS) (acute psychological stress model associated with blood sampling), the serum triglyceride (TG) levels showed a rapid decrease followed by a rebound at 24 h as we have recently reported. However, such a TG rebound was impaired in the CpG/GalN- and solely CpG-treated groups of mice despite an absence of liver injury based on serum alanine aminotransferase levels in the latter group. Thus, the stress-associated serum TG rebound was abrogated by the injection of a sub-hepatotoxic CpG dose. In the second experiment, we simply measured the hepatic CD36 and SACRB1 (the gene for scavenger receptor B1 (SR-B1)) transcripts after the i.p. administration of PBS, CpG or CpG/GalN. There was a remarkable elevation of hepatic CD36 transcript expression in both the CpG- and CpG/GalN-treated mice at 8 h post-CpG injection whereas the increase in the PBS-treated mice was slower than the former two groups, suggesting that hepatic CD36 transcript expression is more pronounced in the combined stress models than under psychological stress alone. The individual mice data showed that the increase in CD36 expression was accompanied by a reduction in SCARB1 mRNA, showing reciprocal regulation between these two genes. Together with our previously reported findings, these data suggest that, in a murine model combining psychological stress with TLR-triggered hepatic inflammation, the psychological stress facilitates liver uptake of plasma TG (and its components fatty acids), but the subsequent re-esterification and/or release of TG-rich lipoproteins from the liver is impaired due to the concomitant TLR-signaling. We hypothesize that lipid metabolism during acute stress shifts toward an elevated hepatic uptake of lipids due to concomitant TLR signaling, facilitating the clearance of bacterial lipids by the liver.
文摘Toll-like receptors (TLRs) recognize specific motifs which are frequently present in bacteria, fungi, prokaryotes and viruses. Amongst TLRs, TLR9 can be activated by such bacterial or viral DNA fragments, immunoglobulin-DNA complexes or synthetic oligonucleotides, which all contain unmethylated cytosineguanine nucleotide sequences (CpGs). Emerging data indicate that TLR9 signaling has a role in, and may influence, colorectal carcinogenesis and colonic inflammation. CpGs are classified into three groups according to their influence on both the antigen-specific humoraland cellular immunity, and the production of type 1 interferons and proinflammatory cytokines. TLR9 activation via CpGs may serve as a new therapeutic target for several cancerous and various inflammatory conditions. Due to its probable anti-cancer effects, the application possibilities of TLR9-signaling modulation may be extremely diverse even in colorectal tumors. In this review we aimed to summarize the current knowledge about TLR-signaling in the pathogenesis and therapy of inflammatory bowel diseases and colorectal cancer. Due to the species-specific differences in TLR9 expression, however, one must be careful in translating the animal model data into the human system, because of the differences between CpG-oligodeoxynucleotide-responsive cells. TLR9 agonist DNA-based immunomodulatory sequences could also represent a promising therapeutic alternative in systemic inflammatory conditions and chronic colonic inflammations as their side effects are not significant.
基金Supported by The Sao Paulo Research Foundation(FAPESP),NO.2013/14022-6 and NO.2014/17716-1
文摘BACKGROUND Toll-like receptors(TLRs)are the first line of host defense,and are involved in Helicobacter pylori(H.pylori)recognition and activation of both inflammatory and carcinogenic processes.The presence of single nucleotide polymorphisms(SNPs)in genes that activate the immune response may modulate the risk of precancerous lesions and gastric cancer(GC).Among them,Toll-like receptor 9(TLR9)polymorphisms have emerged with a risk factor of infectious diseases and cancer,however the studies are still inconclusive.AIM To evaluate whether TLR9 rs5743836 and rs187084 SNPs contribute to the risk of gastric carcinogenesis,and its influence on mRNA expression.METHODS A case-control study was conducted to evaluate two TLR9 SNPs(TLR9-1237 TCrs5743836 and TLR9-1486 CT-rs187084)in chronic gastritis(CG)and GC patients.A total of 609 DNA samples of peripheral blood[248 CG,161 GC,and 200 samples from healthy individuals(C)]were genotyped by polymerase chain reaction-restriction fragment length polymorphism.All samples were tested for the H.pylori infection using Hpx1 and Hpx2 primers.Quantitative polymerase chain reaction by TaqMan?assay was used to quantify TLR9 mRNA from fresh gastric tissues(48 GC,26 CG,and 14 C).RESULTS For TLR9-1237,the TC+CC or CC genotypes were associated with a higher risk of GC than C[recessive model odds ratio(OR)=5.01,95%confidence interval(CI):2.52-9.94,P<0.0001],and the CG(recessive model OR=4.63;95%CI:2.44-8.79,P<0.0001)groups.For TLR9-1486,an association between the CT+TT genotypes and increased risk of both GC(dominant model OR=2.72,95%CI:1.57-4.72,P<0.0001)and CG(dominant model OR=1.79,95%CI:1.15-2.79,P=0.0094)was observed when compared to the C group.Moreover,the presence of TLR9-1237 TC/CC+TLR9-1486 CC genotypes potentiate the risk for this neoplasm(OR=18.57;95%CI:5.06-68.15,P<0.0001).The TLR9 mRNA level was significantly higher in the GC group(RQ=9.24,P<0.0001)in relation to the CG group(RQ=1.55,P=0.0010)and normal mucosa(RQ=1.0).When the samples were grouped according to the polymorphic genotypes and the presence of H.pylori infection,an influence of TLR9-1237 TC+CC polymorphic genotypes(P=0.0083)and H.pylori infection(P<0.0001)was observed on the upregulation of mRNA expression.CONCLUSION Our findings show that TLR9 rs5743836 and rs187084 polymorphisms are associated with a higher risk of carcinogenesis gastric,and that TLR9 mRNA levels can be modulated by TLR9-1237 TC+CC variant genotypes and H.pylori infection.
基金supported by a grant from Natural Sciences Foundation of Hubei Province, China (No. 2006ABA139)
文摘The aim of the present study was to investigate the expression of toll-like receptors （TLR） 9 in peripheral blood mononuclear cells （PBMC） of patients with chronic hepatitis B and C with different virus copies. The study group included 90 patients （60 with chronic hepatitis B, and 30 with chronic hepatitis C）, and 20 healthy people served as control group. The protein and mRNA levels of TLR9 were detected by using flow cytometry and real-time PCR. The serum viral copies of HBV and HCV were measured in all patients, and the correlation between HBV-DNA copies or HCV-RNA copies and the TLR9 expression was analyzed. Our results demonstrated that HBV or HCV infection led to a decreased expression of TLR9 mRNA and protein compared to the control group （P〈0.05）. The TLR9 protein and mRNA levels were negatively correlated with serum viral copies of HBV and HCV （r=-0.632, r=-0.909, P〈0.01）. It was concluded that TLR9 mRNA and protein are down-regulated in PBMC of HBV-infected or HCV-infected patients, and they are negatively correlated with serum viral copies and play an important role in detecting viral replication of HBV and HCV.
基金Supported by(in part)Grants from the Ministry of Education,Culture,Sports,Science and Technology,Japan(to Katakura K)
文摘Abnormal innate immune responses toward luminal bacteria play an important role in the pathogenesis of inflammatory bowel disease.It has been demonstrated that bacteria having CpG DNA ameliorate experimental colitis in mice,and Toll-like receptor 9 (TLR9) signaling mediates the anti-inflammatory effects in mouse colonic inflammation.A gene variation in NOD2/CARD15 has been reported in Crohn's disease (CD) patients in Western countries,but this variation has not been identified in Japanese CD patients.Therefore,we hypothesized that TLR9 is a key factor in the development of ulcerative colitis (UC),and we investigated gene mutations and polymorphisms of TLR9 in Japanese UC patients.Three single nucleotide polymorphisms (SNPs) in TLR9 were identified in healthy controls,and were assessed in 48 UC patients and 47 healthy controls.Control subjects were matched for age,sex and date of blood sampling from among a subgroup of participants.We found that TLR9-1486CC,1174GG and 2848AA increase the risk of UC [odds ratio (OR) 2.64,95% confidence interval (95% CI):1.73-6.53,P=0.042],and TLR9-1486TT,1174AA and 2848GG decrease the risk of UC (OR 0.30,95% CI:0.10-0.94,P=0.039),although there were no correlations between SNPs and disease phenotype or TLR9 mRNA expression.These findings suggest that TLR9 polymorphisms are associated with increased susceptibility to UC.
文摘目的探讨固有免疫信号通路分子IRAK-M在TLR9信号诱导肝组织淋巴细胞浸润过程中的作用。方法应用野生型B6小鼠及IRAK-M基因敲除小鼠,腹腔注射TLR9配体CpG寡核苷酸激活肝内TLR9信号系统,提取小鼠肝内浸润淋巴细胞,并采用流式细胞术检测肝内浸润不同淋巴细胞群的比率及细胞因子的表达情况。结果 CpG ODN激活TLR9信号通路后,IRAK-M基因敲除小鼠肝内浸润CD4+和CD8+T细胞比率较野生型B6小鼠显著增加[CD4+:(22.50±0.73)% vs. (20.03±0.75)%;CD8+:(17.83±0.67)%vs.(15.55±0.75)%;P均<0.05];肝内淋巴细胞细胞因子白细胞介素-6(IL-6)和肿瘤坏死因子(TNF-α)合成增加[IL-6:(1.91±0.26)% vs. (0.93±0.12)%;TNF-α:(13.23±1.23)% vs. (8.16±0.66)%;P均<0.01]。结论 IRAK-M具有负调节肝内TLR9信号系统的作用。
文摘目的通过观察早产儿不同胎龄Toll样受体9(TLR9)的表达,探讨早产儿免疫功能低下的机制。方法采集2010年7月至2014年6月在上海市嘉定区妇幼保健院产科出生的活产新生儿的脐血229份,按胎龄分为4组,28~31周组,31~34周组,34~37周组,≥37周组,采用流式细胞术和实时荧光定量PCR方法,分别检测其TLR9的蛋白和m RNA表达情况,了解其与胎龄之间的关系,并分析m RNA和蛋白表达间的相关性。结果 TLR9阳性细胞率在28~31周组,31~34周组,34~37周组,≥37周组分别为(15.93±6.23)%,(11.63±6.70)%,(13.66±6.88)%,(20.51±12.06)%;其在胎龄28~31周较高,至31~34周逐渐下降至最低,两组差异有统计学意义(P<0.05);34~37周后TLR9阳性细胞率表达逐渐升高,至≥37周达最高,两胎龄组比较,差异具有统计学意义(P<0.05)。31~37周间新生儿脐血TLR9阳性细胞率与胎龄呈正相关(r=0.273,P=0.006)。TLR9 m RNA表达在28~31周组,31~34周组,34~37周组,≥37周组分别为(4.95±3.44)%,(8.89±8.49)%,(13.91±10.92)%,(7.19±7.11)%;其在28~36周逐渐升高,与胎龄呈正相关(r=0.355,P<0.001)。≥37周TLR9 m RNA表达量下降,该值虽高于28~31周,但差异无统计学意义(P>0.05)。相关性分析表明,同胎龄时期同样本新生儿的TLR9 m RNA和TLR9阳性细胞率之间存在负相关(r=-0.227,P=0.011)。结论 TLR9阳性细胞率和TLR9 m RNA表达在不同胎龄组新生儿间有差异,TLR9阳性细胞率表达在31~37周间随着胎龄的增加而增加,TLR9 m RNA在28~36周间随着胎龄的增加而增加。
基金Supported by The National Natural Science Foundation of China, No. 30972898
文摘AIM:To determine the expression of toll-like receptor 9(TLR9) in pancreatic tumor and the effects of cytosine phosphate-guanosine oligodeoxynucleotides 2216(CPG ODN2216) on biological behavior of pancreatic carcinoma cell line PANC-1 and explore their clinical significance.METHODS:The immunohistochemistry and Western blot were used to determine the expression of TLR9 protein in pancreatic cancer tissues,and immunofluorescence staining was performed to detect the TLR9 protein expression in pancreatic carcinoma cell line PANC-1.To assess the effects of CPG ODN2216 on the invasive property of Panc-1 cells,in vitro cell adhesion,wound-healing scrape,and invasion and cell colony formation were evaluated.RESULTS:TLR9 was highly expressed in pancreaticcancer tissues and PANC-1 cells.The percentage of positive cells expressing TLR9 protein in human pancreatic tissues,paracancerous tissues and normal tissues were 73.3%,33.3% and 20.0%,respectively,and the protein expression level of TLR9 was gradually descending(P < 0.05).In vitro tests in wound-healing scrape,cell adhesion,colony formation and matrigel invasion showed that the adhesion and motility of PANC-1 cells in CPG ODN 2216 treatment group were signif icantly lower than in the control group(P < 0.05).The cell growth assay showed that the proliferative ability of PANC-1 cells in treatment group was significantly decreased and CPG ODN2216 had an inhibitive effect in the growth of Panc-1 cells in a dose and time-dependent manner(P < 0.05).CONCLUSION:The gene of TLR9 is correlated with the invasive and metastatic potential of human pancreatic carcinoma,and CPG ODN2216 induces the inhibition of migration and invasion of Panc-1 cells.
基金supported by grants from the National Natural Sciences Foundation of China (No.30973184)the "973" Program of China (No. 2009CB521800)
文摘Inflammation and infection play an important role in the pathogenesis of many cancers.Toll-like receptors（TLRs） are a class of pattern recognition receptors that recognize conserved components of microbes and trigger the immune response against invading microorganisms.Toll-like receptor 9（TLR9） recognizes non-methylated cytosine-phosphateguanosine（CpG） DNA sequences which are the surrogate for viral DNA.TLR9 may react to tumor development and progression during chronic inflammation that involves the tumor microenvironment.In order to study the role of TLR9 in cervical cancer,we analyzed the TLR9 expression in different types of HPV infection cervical cancer cells.Then we detected if CpG sequences influenced the TLR9 expression and the sensitivity to cisplatin（DDP） of these cervical cancer cells in vitro.The expression of TLR9 mRNA and protein in SiHa,Hela and C33A cells was detected by RT-PCR and Western blotting.Real-time PCR was used to examine the TLR9 expression changes induced by CpG.Chemosensitivity of the cervical cancer cells to cisplatin（DDP） was measured by MTT.It was observed that the expression of TLR9 mRNA and protein was increased gradually in SiHa（HPV16＋）,Hela（HPV18＋） and C33A（HPV-） cells.Low doses of CpG increased the TLR9 expression only in C33A（HPV-） cells,but not in SiHa（HPV16＋） and Hela（HPV18＋） cells.Furthermore,low dose of CpG significantly increased the sensitivity of C33A（HPV-） cells,but not that of SiHa（HPV16＋） and Hela（HPV18＋） cells.These results indicated that TLR9 may serve as a protective agent in HPV negative cervical cancer cells.It was concluded that TLR9 could improve the sensitivity to DDP in HPV negative cervical cancer cells and might represent a potential therapeutic option in clinical practice.
文摘Klebsiella has been considered as initiator of AS （ankylosing spondylitis） for nearly four decades. This study aimed to demonstrate that Klebsiella triggers ERS （endoplasmic reticulum stress） and HLA-B27 heavy chain misfolding. CA46 cells or splenocytes obtained from wild-type, MyD88/ or TLR9/ mice were stimulated with KP （Klebsiella pneumoniae） or its components including CPS （capsule polysaccharide）, LPS （lipopolysaccharide）, and KP gDNA （genomic deoxyribonucleic acid） respectively for 24 h and 48 h. The activation of ERS-related signaling was detected by Western blotting or RT-PCR, and the level of misfolded HLA-B27 was determined by non-reducing protein gel electrophoresis and Western blotting. The protein expression of BiP/Grp78 and calreticulin, the alternative splicing of XBP-1 mRNA （messenger ribonucleic acid）, and the activation of caspase-12 and p38 were increased in a dose-dependent manner in HLA-B27-expressing CA46 cells after treatment with decapsulated KP. We also demonstrate that the EP, S-inducing effects occur via the TLR （Toll-like receptor）/MyD88-dependent signaling pathway. Significantly, HLA-B27 misfolding was also detected in decapsulated KP-treated B27-expressing cells. These results suggest that the non-antigen-specific induction of ERS and B27 misfoiding through TLR/MyD88 signaling might promote KP antigen-initiated autoreactive responses via the presentation of misfolded B27, and that small-molecules targeting TLRs might have potential as novel therapeutic agents for AS.
