Toll样受体9基因启动子单核苷酸多态性与儿童变应性哮喘的相关性

Objective To investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) in the promoter region of the toll-like receptor 9 gene (TLR9)in Chinese Han children from Zhejiang province, and their associations with asthma susceptibility and phenotypes. Methods A case-control study was conducted. A total of 312 asthmatic children aged between 1.9 and 11.6 and 339 age matched healthy controls were enrolled in this study from April 2007 to November 2008. The -1486 C/T in rs187084 and -1237 C/T in rs5743836 loci of the TLR9 gene were genotyped by direct DNA sequencing of the PCR products. Serum levels of IFN-γ, IL-12 and IL-4 were detected by enzyme linked immunosorbent assay. Serum levels of total IgE were detected by chemiluminescence, and serum levels of ildren (P＜0.01). The CC genotype had the lowest levels of serum IFN-γand the highest levels of serum IL-4 among the three genotypes. There were no significant differences in these cytokines among the healthy controls (P＞0.05). No statistical differences of serum IL-12 were found among the three genotypes in the two groups (P＞0.05). (4) There were no significant differences of total IgE (log-transformed) among the three genotypes in the asthmatic children (P＞0. 05). Conclusion The -1237 C/T polymorphism of TLR9 gene was not detected in Chinese Han children in this study. The - 1486 C/T polymorphism was associated with the levels of serum IFN-γ and IL-4 in children with asthma.However, there were no correlations between the -1486C/T polymorphism and serum IL-12 levels, total IgE levels or asthmatic susceptibility.     

Toll样受体9基因启动子单核苷酸多态性与儿童变应性哮喘的相关性

Objective To investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) in the promoter region of the toll-like receptor 9 gene (TLR9)in Chinese Han children from Zhejiang province, and their associations with asthma susceptibility and phenotypes. Methods A case-control study was conducted. A total of 312 asthmatic children aged between 1.9 and 11.6 and 339 age matched healthy controls were enrolled in this study from April 2007 to November 2008. The -1486 C/T in rs187084 and -1237 C/T in rs5743836 loci of the TLR9 gene were genotyped by direct DNA sequencing of the PCR products. Serum levels of IFN-γ, IL-12 and IL-4 were detected by enzyme linked immunosorbent assay. Serum levels of total IgE were detected by chemiluminescence, and serum levels of ildren (P＜0.01). The CC genotype had the lowest levels of serum IFN-γand the highest levels of serum IL-4 among the three genotypes. There were no significant differences in these cytokines among the healthy controls (P＞0.05). No statistical differences of serum IL-12 were found among the three genotypes in the two groups (P＞0.05). (4) There were no significant differences of total IgE (log-transformed) among the three genotypes in the asthmatic children (P＞0. 05). Conclusion The -1237 C/T polymorphism of TLR9 gene was not detected in Chinese Han children in this study. The - 1486 C/T polymorphism was associated with the levels of serum IFN-γ and IL-4 in children with asthma.However, there were no correlations between the -1486C/T polymorphism and serum IL-12 levels, total IgE levels or asthmatic susceptibility.     

IFN-γ与 IL-4基因多态性与儿童哮喘易感性及外周血IFN-γ、IL-4和IgE的关系

目的: 探讨IFN-γ和 IL-4 基因多态性与儿童哮喘易感性及血浆IFN-γ、IL-4和IgE的相关性。方法: 用聚合酶链反应-限制性酶切片段长度多态性(PCR-RFLP)法检测100例哮喘儿童和122例对照儿童IFN-γ基因-179G/T、 IL-4 基因-33C/T和-589C/T位点基因型;等位基因特异性-聚合酶链反应(AS-PCR)法检测IFN-γ基因+874A/T位点基因型;毛细管电泳法检测IFN-γ基因CA重复序列基因型;ELISA法测定血浆IFN-γ、IL-4和IgE。结果: 100例哮喘儿童和122例对照儿童IFN-γ基因-179位点均为GG纯合子,未检测到突变基因型。IFN-γ基因+874A/T位点和CA重复序列的基因型和等位基因频率分布在哮喘组和对照组间无显著差异(P>0.05);+874位点多态性与血浆IFN-γ水平相关,AA基因型IFN-γ含量低于AT基因型(P<0.05)。 IL-4 基因-33C/T和-589C/T位点的基因型和等位基因频率分布在哮喘组和对照组间有显著差异(P<0.05);-33和-589位点TT基因型外周血IL-4和总IgE浓度均高于CT基因型,只有-33位点与血浆IL-4水平存在相关性(P<0.05)。结论: IFN-γ基因+874A/T和CA重复序列多态性可能与儿童哮喘无相关性,+874A/T位点多态性与IFN-γ水平相关。 IL-4 基因-33TT和-589TT基因型可能为儿童哮喘的易感基因型,-33位点多态性与IL-4表达水平相关。     

TLR9多态性和IFN-γ表达与结直肠癌的相关性

目的 探讨Toll样受体9(Toll-like receptor 9,TLR9)-1237T/C、-1486T/C和2848 G/A多态性和IFN-γ表达与四川结直肠癌相关性.方法 246名结直肠癌患者和246名健康对照个体TLR9多态性位点基因型采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)分析,ELISA方法检测结直肠癌组织匀浆中IFN-γ、TGF-β和TNF-α细胞因子蛋白水平.结果 TLR9-1237T/C和2848G/A多态性基因型在病例-对照组中频率无显著性差异,-1486T/C CC基因型显著增加了个体患结直肠癌的风险；IFN-γ在-1486T/C CC基因型个体结直肠癌组织中浓度显著降低,在-1486T/C不同基因型个体组中的表达有显著性差异.结论 TLR9-1486T/C可调节IFN-γ在结直肠癌组织中的表达,与结直肠癌发生相关.     

TLR2和TLR4单克隆抗体对DSS诱导的小鼠结肠炎肠黏膜细胞因子及肠道菌群的影响

Objective To evaluate the effect of TLR2McAb and TLR4McAb on intestinal flora of DSS-induced colitis in mice. Methods Fifty healthy male BALB/c mice (SPF level), were randomly assigned into five groups: the control group( group A), the UC model group( group B), TLR2McAb intervention group( group C), TLR4McAb intervention group( group D) and TLR2McAb + TLR4McAb intervention group(group E). Clinical symptoms were evaluated by the disease activity index(DAI), while tissue sam ples were evaluated by histological scoring(HS). The quantities of mRNA for IFN-γ, IL-4 and IL-17 were determined by real-time PCR. Meanwhile, fecal samples were obtained directly from the cecum for microbiological studies. Results After the treatment with TLR2McAb and TLR4McAb, DAI and HS were decreased significantly. Compared with group A, inflammatory cytokines such as IFN-γ, IL-4 and IL-17 in group B were higher. Compared with group B, expression of these three cytokines in group C to E was all markedly decreased. Group A showed a considerable predominance of Lactobacillus spp and Bifidobacterium spp,while the UC model group showed a conspicuous increase of Escherichia coli and decreases of Lactobacillus spp and Bifidobacterium spp. After treatment with TLR2McAb or/and TLR4McAb, Lactobacillus spp and Bifidobacterium spp increased to the normal level. But counts of E. Coli in the three intervention groups were not changed. Conclusion TLR2McAb and TLR4McAb suppressed the development of DSS-induced colitis and increase cecum counts of Lactobacilli and Bifidobacteria.     

TLR2和TLR4单克隆抗体对DSS诱导的小鼠结肠炎肠黏膜细胞因子及肠道菌群的影响

Objective To evaluate the effect of TLR2McAb and TLR4McAb on intestinal flora of DSS-induced colitis in mice. Methods Fifty healthy male BALB/c mice (SPF level), were randomly assigned into five groups: the control group( group A), the UC model group( group B), TLR2McAb intervention group( group C), TLR4McAb intervention group( group D) and TLR2McAb + TLR4McAb intervention group(group E). Clinical symptoms were evaluated by the disease activity index(DAI), while tissue sam ples were evaluated by histological scoring(HS). The quantities of mRNA for IFN-γ, IL-4 and IL-17 were determined by real-time PCR. Meanwhile, fecal samples were obtained directly from the cecum for microbiological studies. Results After the treatment with TLR2McAb and TLR4McAb, DAI and HS were decreased significantly. Compared with group A, inflammatory cytokines such as IFN-γ, IL-4 and IL-17 in group B were higher. Compared with group B, expression of these three cytokines in group C to E was all markedly decreased. Group A showed a considerable predominance of Lactobacillus spp and Bifidobacterium spp,while the UC model group showed a conspicuous increase of Escherichia coli and decreases of Lactobacillus spp and Bifidobacterium spp. After treatment with TLR2McAb or/and TLR4McAb, Lactobacillus spp and Bifidobacterium spp increased to the normal level. But counts of E. Coli in the three intervention groups were not changed. Conclusion TLR2McAb and TLR4McAb suppressed the development of DSS-induced colitis and increase cecum counts of Lactobacilli and Bifidobacteria.     

天津地区汉族人群IL-10-592C/A基因多态性与冠状动脉支架术后再狭窄的关系

目的 探讨中国天津地区汉族人群白细胞介素-10(interleukin-10,IL-10)-592C/A基因多态性的功能性以及其对经皮冠状动脉介入治疗(percutaneous coronary intervention,PCI)术后再狭窄的发病,PCI术后血清IL-10水平的影响.方法 对437例接受PCI并进行冠状动脉造影随访的患者,按冠状动脉造影结果分为再狭窄组(166例)和非再狭窄组(271例),应用聚合酶链反应-限制性片段长度多态性方法检测IL-10-592位点基因型和等位基因频率的分布;用酶联免疫吸附试验法测定2组PCI术前及PCI术后24 h血清IL-10浓度,并比较两组间和各基因型间IL-10水平.结果 (1)IL-10-592C/A基因型和等位基因频率在再狭窄组和非再狭窄组之间差异无统计学意义(P均＞0.05);(2)PCI术后24 h血清IL-10水平再狭窄组显著低于非再狭窄组[(82.67±35.02)ng/Lvs.(95.08±32.26)ng/L,P＜0.05];(3)IL-10-592位点A等位基因携带者(AA+AC基因型)术后24 h血清IL-10水平明显低于非携带者(CC型)[(86.13±34.77)ng/L vs.(102.50±27.52)ng/L,P＜0.05];(4)再狭窄组A等位基因携带者术后24 h血清IL-10水平明显低于非携带者[(78.51±34.09)ng/L vs.(102.19±33.66)ng/L,P＜0.05];(5)再狭窄危险的多因素Logistie回归分析显示:急性冠状动脉综合征、术前狭窄程度、靶病变长度与冠状动脉内支架再狭窄呈正相关(()R值分别为5.90、1.86、2.83),术后24 h血清IL-10水平、参照血管直径、支架直径与冠状动脉内支架再狭窄呈负相关(OR值分别为0.99、0.70、0.46).结论 (1)IL-10基因-592 C/A多态性与中国天津地区汉族人群再狭窄发病无关;(2)IL-10是PCI术后早期的炎症细胞因子,术后24 h血清IL-10水平为再狭窄的独立预测因素,携带A等位基因的个体可能通过降低其表型血清IL-10水平而增加了冠状动脉内支架术后再狭窄的发病.

Abstract：

Objective To investigate the relationship of interleukin-10 gene (IL-10)polymorphism and the serum IL-10 level with restenosis after percutaneous coronary intervention (PCI) in Tianjin Chinese Han population and study the effect of IL-10 gene polymorphism on serum IL-10 level. Methods Four hundred and thirty-seven patients who successfully underwent PCI with a follow-up angiography were divided into a restenosis group (n= 166) and non-restenosis group (n= 271). The IL-10 gene promoter polymorphism at position -592 was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Meanwhile their serum IL-10 level before and 24 h after PCI was determined by enzyme-linked immunosorbent assay (ELISA). Results (1) There was no significant difference in frequencies of -592 genotypes and alleles between the two groups (P＞0. 05); (2) The 24 hpost-PCI IL-10 serum level of restenosis group was significantly lower than that of the non-restenosis group [(82. 67±35. 02) ng/L vs. (95.08±32.26) ng/L, P＜0.05]; (3) The serum level of the A allele carriers (AA+AC) was significant lower than that of the CC carriers [(86.13±34.77) ng/L vs. (102. 50±27.52)ng/L,P＜0.05]; (4) In the restenosis group, the 24 h post-PCI serum level of IL-10 in the A allele carriers was also significantly lower than that in those without the A allele [(78.51 ± 34.09) ng/L vs. (102.19 ±33.66) ng/L, P＜ 0. 05]; (5) Logistic regression analysis revealed positive correlations between acute coronary syndrome patients, pre-PCI degree of stenosis, length of target stenosis lesion and restenosis (OR=5.90, 1.86, 2.83 respectively); and there were negative correlations between 24 h post-PCI serum level of IL-10, the stent diameter, the diameter of reference vessel before stent implantation and restenosis (OR=0. 99, 0. 70, 0. 46 respectively). Conclusion (1) TheIL-10 gene -592 C/A polymorphism was not associated with restenosis in the Tianjin Chinese Han population; (2) IL-10 is an early post-PCI inflammatory cytokine, 24 h post-PCI serum IL-10 level was an independent predictive factor for restenosis,the IL-10 A allele carriers may have increased incidence of in-stent restenosis (ISR) by reducing the serum IL-10 levels.     

Toll-like receptors and CD14 genes polymorphisms and susceptibility to asthma in Tunisian children

Many studies have shown the implication of CD14 and toll-like receptors (TLRs) 2, 4 and 9 in the pathogenesis of asthma or atopy. To evaluate the association of CD14 and TLRs gene polymorphisms with asthma or atopy, 210 asthmatic children, 224 controls and 80 families were enrolled in this study. Six single nucleotide polymorphisms TLR2 (+2408 G-->A), TLR4 (+1196 C-->T), TLR4 (+896 A-->G), TLR9 (-1237 T-->C), TLR9 (-1486 T-->C) and CD14 (-159 C-->T) were genotyped using polymerase chain reaction followed by restriction fragment length polymorphism in the case-control and family study. The -1237C allele in TLR9 gene polymorphisms was associated with increased risk of asthma [odds ratio 1.53, 95% confidence interval (1.03-2.27)], although no statistically significant differences in allele or genotype frequencies of four other TLRs polymorphisms were evident between the asthmatic and control groups. The CD14 -159 C allele was found to be significantly higher in the asthmatic group when compared with controls (P=0.0006<0.05). Transmission disequilibrium test of 80 asthmatic families showed significant transmission of the -159 C allele in the CD14 gene to asthma-affected offspring. It was concluded that TLR9 and CD14 gene polymorphisms may contribute to an inherited predisposition to asthma in Tunisian children.     

The effect of polymorphisms at the CD14 promoter and the TLR4 gene on asthma phenotypes in Turkish children with asthma

BACKGROUND: Endotoxin, with its potential to enhance type 1 immunity, is a significant player in the hygiene hypothesis. The combined effects of the genetic variants of various molecules in the endotoxin response pathway on asthma related phenotypes are largely unknown. OBJECTIVE: To investigate the effects of the genetic variants of CD14 and TLR4 genes on asthma phenotypes in a large number of asthmatic children. METHODS: 613 asthmatic children were genotyped at the CD14-C159T, TLR4-A896G and TLR4-C1196T loci. IgE, eosinophil numbers and FEV1 were compared in 327 children who were not on any controller medications and were symptom free. Multivariate logistic regression was used to determine the factors associated with total IgE. RESULTS: Among children with atopic asthma, total IgE levels were significantly different among the three genotypes in the co-dominant model [CC: 435 kU/l (interquartile range: 146-820); CT: 361 (140-710); TT 204 (98-435), P = 0.035]. TT genotype was significantly and independently associated with lower IgE levels (OR: 0.5 95%; CI = 0.28-0.90, P = 0.021). Both TLR4-A896G and TLR4-C1196T polymorphisms were more frequent in the mild asthma group with atopy (P = 0.032, 0.018, respectively). The combined effects of the genetic variants in CD14 and TLR4 genes did not improve the observed associations. CONCLUSION: Our study demonstrates that the CD14-C159T promoter variant influences total IgE levels and also indicates that the T allele has a more profound effect on total IgE in children with atopic asthma. Polymorphisms in the TLR4 gene may be associated with milder forms of disease in atopic asthmatics in the population studied.     

TLR-9 promoter polymorphisms (T-1237C and T-1486C) are not associated with systemic lupus erythematosus: A case control study and meta-analysis

Toll like receptors (TLRs) are essential molecules implicated in both innate and adaptive immune response. Polymorphisms in TLR gene have been associated with various infectious diseases and autoimmune disorders. Role of TLR9 has been elegantly demonstrated in both human systemic lupus erythematosus (SLE) and mice model of lupus. In the present study we investigated association of TLR-9 promoter polymorphisms (T-1237C and T-1486C) with susceptibility/resistance to SLE in an Eastern Indian state which is endemic to parasitic diseases. 210 Female SLE patients who fulfilled the American College of Rheumatology criteria were enrolled along with matched healthy controls from Odisha, India. TLR-9 polymorphisms (T-1237C and T-1486C) were typed by polymerase chain reaction followed by restriction fragment length polymorphism. For meta-analysis, relevant literatures were searched from PubMed database and comprehensive meta-analysis V2 software was employed for analysis. Allele and genotype frequency of TLR-9 promoter polymorphisms (T-1237C and T-1486C) were comparable among SLE patients and controls. Further, meta-analysis of earlier reports and present study did not reveal a significant association of TLR-9 (T-1237C and T-1486C) polymorphisms with SLE. Data from the present study suggest that TLR-9 promoter polymorphisms are not associated with susceptibility to SLE in an area endemic to parasitic diseases.     

呼吸道合胞病毒毛细支气管炎患儿IL4/IFNγ水平变化及意义

为研究Th1/Th2类细胞因子 (CK )在呼吸道合胞病毒毛细支气管炎 (RSV毛支 )发病中的作用 ,本文采用酶联免疫吸附试验 ,对 2 0例RSV毛支患儿和 15例健康婴幼儿外周血单个核细胞 (PBMC )经植物血凝素 (PHA )刺激培养后 ,测定上清液中某些细胞因子 ,并同时检测患儿血浆总IgE水平。实验结果表明 :(1)经PHA刺激培养后 ,RSV毛支组上清液中IL 4,IFN γ水平及IL 4/IFN γ比值与对照组比较 ,差异有显著性 (P <0 0 1、 0 0 5、 0 0 1)。RSV毛支组血浆总IgE水平较对照组明显升高 (P <0 0 1) ;(2 )相关分析 :培养上清液中IL 4水平 ,IL 4/IFN γ比值与血浆总IgE水平间呈显著正相关 ,IFN γ与IgE间呈显著负相关。此外IL 4与IFN γ间呈显著负相关。结果提示 ,RSV毛支患儿存在Th1/Th2功能紊乱 ,主要表现为Th2细胞功能亢进 ;IL 4、IFN γ参与了RSV感染时的免疫病理机制。     

Association study of Toll-like receptor 9 gene polymorphism in Korean patients with systemic lupus erythematosus

Mammalian Toll-like receptors (TLR) play an important role in both adaptive immunity and innate immunity. Genetic variations within TLR genes are known to be associated with a variety of inflammatory and infectious diseases. TLR9 is potentially associated with autoimmune diseases, because it participates in the production of pro-inflammatory cytokines and the maturation of dendritic cells. We investigated the association of four TLR9 gene polymorphisms (-1486 T>C, -1237 C>T, +1174 A>G and +2848 G>A) with the susceptibility to systemic lupus erythematosus (SLE) and related phenotypes in 680 Korean people (350 SLE patients and 330 controls). TLR9 gene polymorphisms were not significantly associated with the susceptibility to SLE and related phenotypes.     

血清IL-33、IFN-γ与IgE在支气管哮喘患者中的表达及临床意义

目的观察血清白细胞介素-33(IL-33)、干扰素-γ(IFN-γ)及IgE在支气管哮喘患者中的表达及意义。方法选取94例急性发作期支气管哮喘患者,同时选取同期60例健康体检的人群为对照组,采用酶联免疫法测定血清中IgE、IL-33及IFN-γ水平。比较两组IgE、IL-33和IFN-γ的水平。分析IL-33、IFN-γ和IgE的相关性。结果支气管哮喘患者外周血中IFN-γ水平低于正常人群(t=4.533,P<0.001);IL-33、IgE水平高于正常人群(t=5.831、66.129,P<0.001,<0.001),差异有统计学意义。IgE水平与IL-33水平呈正相关(r=0.667,P=0.032),IFN-γ与IgE、IL-33呈负相关(r=-0.714,P=0.024;r=-0.623,P=0.038)。结论血IgE、IL-33和IFN-γ水平的变化在支气管哮喘患者发病过程中起到一定的作用。