甘肃不同地区绵羊TLR9基因多态性分析

针对由于急、慢性呼吸道引起的生产性能降低甚至死亡,选择与该病相关的基因TLR9为研究对象,分析群体中该基因的遗传多态性及变异特征,为进一步揭示舍饲绵羊的遗传特性和生产利用提供基础数据。采用PCR-SSCP检测427只表型正常和58只患呼吸道疾病的舍饲绵羊TLR9基因的多态性,测序群体内变异的各等位基因数列,并构建系统发育树以明确舍饲绵羊TLR9基因等位基因之间的遗传关系。结果显示,甘肃地区绵羊在TLR9基因中发现了4个等位基因,共7个核苷酸多态位点,这些位点多是由点突变形成,其中转换4个(占57.14%),颠换3个(占42.86%)。甘肃地区绵羊TLR9基因具有较丰富的多态性;TLR9基因多态性与绵羊呼吸道疾病有一定的相关性。     

动物TLR4基因多态性及其抗病相关性研究进展

TLR4是天然免疫系统中的一种重要模式识别受体,可选择性识别病原微生物而启动天然免疫,在宿主天然免疫和获得性免疫中具有重要作用。此外,大量研究表明,TLR4基因的多态性与多种动物的细菌、病毒和寄生虫性疾病密切相关,说明通过TLR4基因的研究可为动物分子标记辅助选育研究提供依据。该文简要论述部分动物TLR4基因多态性及其与抗病相关性的研究进展。     

TLR4基因启动子区多态性及其对蛋白表达的影响

目的:检测中国人群Toll样受体4(Toll-like receptor 4,TLR4)基因5′调控区的单核苷酸多态性(single nucleotidepolymorphisms,SNPs),探讨其与TLR4蛋白表达的关系.方法:采用聚合酶链反应-限制性片段长度多态性法(PCR-RFLP)对正常汉族人群样本TLR4启动子区-2242、-1892和-1837这3个可能有意义的SNP位点进行基因分型,以确定中国人群中TLR4基因启动子区SNP基因型和发生频率.取其中89例全血标本用全血培养模型检测内毒素刺激前后TLR4蛋白的表达变化,进一步探讨TLR4启动子区单核苷酸多态性对其蛋白表达的影响.结果:TLR4启动子区-2242、-1892和-1837这3个可能有意义的SNP住点等位基因频率分别是43.27%、27.70%和42.75%.TLR4蛋白表达检测结果表明内毒素刺激后-2242位点TC与CC基因型TLR4蛋白的表达显著高于TT基因型(P<0.05),其它位点则没有影响.结论:中国汉族人群中TLR4基因启动子区-2242位点可能是脓毒症关联分析重要的遗传标记.     

TLR4 基因启动子单核苷酸多态性与结直肠癌临床预后的关系

目的:探讨Toll样受体(TLR)基因启动子单核苷酸多态性(SNP)与结直肠癌临床预后的关系。方法:收集我院2006年1月到2010年1月收治的结直肠癌患者200例,通过PCR扩增外周血DNA,经过查找数据库发现TLR4基因启动子区域有rs137853920、ss77136219多态位点,对所有患者随访5年,比较不同手术方式、美国癌症联合委员会(AJCC)分期、分化程度患者的总生存时间(OS)率和疾病无进展时间(PFT)率,分析基因型频率以及单倍体频率对患者生存影响。结果:200例患者生存时间在4~60个月,中位生存时间为54个月,OS率和PFT率在不同手术方式、癌症AJCC分期、分化程度患者间差异均具有统计学意义(P0.05);对生存资料进行多因素的Cox回归分析,结果显示rs137853920基因多态位点基因型AA、AG、GG具有较好的预后(P0.05),而ss77136219基因多态位点基因型GG、GA、AA具有较差预后(P0.05)。rs137853920、ss77136219多态位点共有四个单倍体型分别为AA、AG、GA、GG,频率分别为26.3%、21.7%、38.5%、21.6%,经过Cox多因素分析AG型患者具有较好预后(P0.05),而GG型患者具有较差预后(P0.05)。结论:TLR4基因启动子SNP可以作为结直肠癌临床预后的特异指标。     

基质金属蛋白酶-9 基因单核苷酸多态性与肺结核病的相关性研究

目的:探讨基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)基因多态性(single nucleotide polymorphism,SNP)与肺结核的相关性。方法:对符合纳入及排除标准的肺结核病例组224例及健康对照组249例进行血样收集与临床资料采集。采用飞行时间质谱分析方法对MMP-9基因rs17576、rs2236416、rs3787268、rs3918254共4个多态性位点进行基因分型,数据统计分析采用SPSS20.0和Haplo View 4.0软件进行。结果:我们首次发现,在病例及对照组中,rs17576基因型频率分布存在统计学差异(X~2=7.822,P=0.020)。与对照组相比,病例组G等位基因频率显著高于对照组(X~2=7.335,P=0.007,OR=1.463,95%CI=1.110-1.927)。病例组rs17576基因型分布中,GG和AG基因型患者吸烟史显著高于AA基因型患者;GG和AG基因型患者卡介苗接种史显著低于AA基因型患者。连锁不平衡分析发现一个单倍型(rs17576-rs3918254)高度连锁(D'0.7;r~20.8)。在病例组及对照组中,G-C和A-C单倍型频率分布存在显著性差异,病例组中G-C单倍型频率显著高于对照组(P=0.022),对照组中A-C单倍型频率显著高于疾病组(P=0.024)。结论:MMP-9基因rs17576多态性位点可能与肺结核有关,携带有rs17576位点G等位基因的个体更易发生肺结核。携带G-C(rs17576-rs3918254)单倍型的个体更易患肺结核病,携带A-C(rs17576-rs3918254)单倍型的个体相对不易患肺结核病。     

绵羊H-FABP基因单核苷酸多态性的研究

以小尾寒羊(48只)、宁夏滩羊(121只)、滩寒杂交羊F1(23只)、无角陶塞特(48只)、萨福克(24只)5个绵羊群体为实验材料, 利用PCR-SSCP和DNA测序技术对心脏型脂肪酸结合蛋白(H-FABP)基因(GenBank登录号: AY157617)外显子2和内含子2部分序列进行单核苷酸多态性(SNPs)检测及遗传多态性分析。结果表明: (1)引物2的PCR扩增产物中存在981(G/A)、1014(A/C)、1019(T/C) 和1058 (-/G ) 4个SNP位点, 表现为AA、BB、CC、AB、AC、BC、AD、CD和BD 9种基因型, 其中AA为优势基因型。经χ2检验后, 除滩羊和萨福克羊外, 其他群体的基因频率和基因型频率均处于Hardy-Weinberg平衡状态。群体遗传多态性分析表明: 宁夏滩羊、小尾寒羊和无角陶塞特羊3个群体中的多态信息含量(PIC)均处于0.25和0.50之间, 为中度多态, 萨福克羊和滩寒杂交羊F1为低度多态(PIC<0.25), 表明脂肪酸结合蛋白基因在不同绵羊品种中具有单核苷酸多态性, 可以进一步作为候选基因来分析其与肌内脂肪含量性状的关联性。(2)引物4的PCR扩增产物中检测到1个SNP多态位点为2407(T/C), 表现为HH、Hh和 hh 3种基因型, 基因型频率大小为HH＞Hh＞hh, 经χ2检验后, 在滩羊和无角陶塞特羊中均为达到Hardy-Weinberg平衡状态, 其多态信息含量均为低度多态(PIC<0.25), 而在小尾寒羊、滩寒杂交羊F1和萨福克羊均没有多态出现。     

TLR9介导CpG激活免疫细胞

机体的天然免疫 (ｉｎｎａｔｅｉｍｍｎｕｎｉｔｙ)在抵抗微生物感染中发挥重要作用。然而机体天然 (非特异性 )免疫系统识别病原微生物的机制尚未完全阐明。近来关于Ｔｏｌｌ样受体(ＴＬＲ)的研究表明 :病原微生物通过呈现多种具有保守序列的分子格局 (ｍｏｌｅｃｕｌａｒｐａｔ ｔｅｒｎ)被机体免疫细胞表面的格局识别受体(ｐａｔｔｅｒｎ ｒｅｃｏｇｎｉｔｉｏｎｐａｔｔｅｒｎ)识别 ,从而诱导天然免疫应答。ＴＬＲ家族是在进化中由昆虫到人类均保守的格局识别受体[1] 。而新近发现的ＴＬＲ9可能是哺乳类细胞识别细菌ＤＮＡ中免疫刺激序…     

中国汉族女性TLR7基因主要编码区的全长扩增及多态性分析

目的:扩增TLR7主要编码区外显子3(exon 3)的全长基因片段并进行基因多态性分析,筛选健康人群中TLR7基因的主要SNP位点.方法:采用酚-氯仿抽提方法从28例健康女性血样中提取基因组DNA,采用长片段扩增方法分别扩增TLR7 exon 3区的3个片段,测序、拼接后进行多态性分析.结果:采用LA-Taq酶体系成功扩增TLR7 exon 3区基因片段;经与Genbank数据库中TLR7参考序列比较,我国女性TLR7基因序列高度保守,仅出现了4个点突变,并发现1个SNP位点RS3853839,表现为GG、CG和CC三种基因型.结论:建立了TLR7编码区基因扩增方法,筛选到1个TLR7SNP位点RS3853839,可为分析TLR7多态性与多种病毒感染性疾病的关系提供参考.     

TLR3和TLR4基因多态性与EV71感染手足口病严重性及易感性的关系研究

手足口病(Hand-foot-and-mouth disease,HFMD)是5岁以下婴幼儿常见的病毒性肠道传染病,该病主要由人肠道病毒71型(Enterovirus 71,EV71)型及柯萨奇A组16型(Coxsackievirus A16,CV-A16)引起,但关于TLR3、TLR4基因多态性与EV71感染手足口病的报道较少。为探讨EV71感染手足口病患儿TLR3和TLR4基因多态性与EV71感染手足口病严重性及易感性的关系,本研究选择2016年8月至2017年8月就诊于安徽医科大学第一附属医院的EV71感染手足口病患儿166例,其中重症组76例,轻症组90例,并选择同期来院体检的健康者120例作为对照组。收集患者入院时的年龄、性别、发热天数等基线资料,采集血液检测白细胞计数(White blood cell count,WBC)、丙氨酸转氨酶(Alanine aminotransferase,ALT)、谷草转氨酶(Glutamate transaminase,AST)、酶联免疫吸附试验(Enzyme-linked immunosorbent assay,ELISA)检测血清C反应蛋白(C reactive protein,CRP)、干扰素-γ(Interferon-γ,IFN-γ)水平;分离外周血单个核细胞提取DNA,琼脂糖凝胶电泳检测DNA情况;聚合酶链反应(Polymerase chain reaction,PCR)扩增TLR3c.1377C/T和TLR4-896A/G,限制性内切酶Tap I(TthHB8 I)、Nco I分别酶切TLR3、TLR4 PCR扩增产物,凝胶成像系统记录实验结果,对扩增产物进行测序,分析其基因多态性结果。结果显示,对照组与EV71感染组TLR3c.1377C/T位点的基因型分布与C、T等位基因频率均无显著统计学意义(P0.05);EV71感染组中重症组TT基因型较轻症组显著升高(P0.05);重症组T等位基因频率显著高于轻症组(P0.05),C等位基因频率显著低于轻症组(P0.05);EV71感染组中,TLR3c.1377C/T位点不同基因型患儿在年龄、性别及ALT、AST、CKMB水平上无显著差异(P0.05);TLR3c.1377C/T位点TT型患儿的发热时间及WBC、CRP水平显著高于CT和CC型,CT型患儿的发热时间及WBC、CRP水平显著高于CC型(P0.05);CC型患儿的IFN-γ水平显著高于CT和TT型(P0.05);TLR4-896A/G基因电泳条带为140bp的特异性扩增产物,为野生型Asp/Asp基因型,对照组和EV71感染组均未出现A→G的突变。本研究得出结论,TLR3c.1377C/T位点有CC、TT、CT三个基因型,且携带T等位基因EV71感染手足口病患儿进展为重症的风险较高;TLR4-896A/G基因无突变,与EV71感染手足口病患儿疾病严重性和易感性无关。     

TNRC9 基因rs12443621A/G 多态性与中国女性乳腺癌易感性及临床病理关系的研究

目的:研究TNRC9/LOC643714 基因 rs12443621A/G 多态性与乳腺癌易感性及临床病理之间的关系.方法:DNA 试剂盒提取321 例乳腺癌患者和340 例正常女性静脉血全基因组DNA,PCR扩增目的基因片段,提取扩增样本进行DNA 测序检测分析rs12443621 多态性.应用 SPSS17.0 软件对实验结果进行统计学分析.结果:应用 SPSS17.0软件对TNRC9/LOC643714 基因rs12443621A/G多态性AA、AG、GG 进行卡方检验分析,结果显示三种基因型分布在病例组及对照组中无统计学意义(X<'2>=1.43,P>0.05),与乳腺癌易感性无关,与乳腺癌病理分型、ER、PR、HER-2状态以及淋巴结是否转移无相关性(X<'2>=2.90,P>0.05;X<'2>=2.25,P>0.05;X<'2>=1.671,P>0.05;X<'2>=1.34,P>0.05;X<'2>=3.24,P>0.05).结论:TNRC9 基因rs12443621A/G 多态性与乳腺癌易感性及临床病理特征无关.不能作为独立的基因标志物对乳腺癌进行早期检测和诊断.

Abstract：

Objective: To investigate the relationship between the rs12443621 polymorphisms of TNRC9/LOC643714 and breast cancer risk and clinico-pathological characteristics in Chinese women. Methods: Genomic DNA was extracted from peripheral blood. The Single Nucleotide Polymorphisms of the TNRC9/LOC643714 rs12443621,from breast cancer of 321 cases and 340 controls were detected by polymerase chain reaction (PCR) and direct DNA sequencing.Results:The genotypes of rs12443621 can not increase the risk for breast cancer(X2=1.43,P>0.05).There was no relationship between the genotypes and pathological category,lymph node metastases, ER status or PR staus or Her-2 status (x2=2.90,P>0.05; X2=2.25,P>0.05;X2=1.671,P>0.05; X2=1.34,P>0.05; X2=3.24,P>0.05). Conelusion:There was no relationship between three genotypes of rs12443621A/G and individual susceptibitity or clinic pathological characteristics of breast cancer.     

绵羊GDF9基因PCR-SSCP分析

生长分化因子9（GDF9）是由卵母细胞分泌的一种生长因子,它对早期卵泡的生长和分化起重要的调节作用。采用PCR－SSCP技术分析了GDF9基因在小尾寒羊、湖羊、多赛特羊和萨福克羊4个绵羊品种的多态性。结果表明：GDF9基因在两对引物扩增片段中均存在PCR－SSCP多态性。对于引物1扩增片段,4个绵羊品种均检测到AA基因型,AB基因型只出现在湖羊、多赛特羊和萨福克羊中,仅在萨福克羊中检测到BB基因型;在4个绵羊品种中,A等位基因频率明显高于B等位基因频率。对于引物2扩增片段,4个绵羊品种均检测到AA基因型,AB基因型只出现在湖羊、多赛特羊和萨福克羊中,4个绵羊品种均没有检测到BB基因型;在4个绵羊品种中,AA基因型频率最高,A等位基因频率明显高于B等位基因频率。引物1的多态性片段测序分析表明：位于GDF9基因cDNA第152处发生了单碱基的改变（A→G）,并导致了氨基酸的改变（天冬酰胺→天冬氨酸）。     

Biased distribution of single nucleotide polymorphisms (SNPs) in porcine Toll-like receptor 1 (TLR1), TLR2, TLR4, TLR5, and TLR6 genes

Toll-like receptors (TLRs) recognize various microbial components and induce immune responses. Polymorphisms in TLRs may influence their recognition of pathogen-derived molecules; swine TLRs are predicted to be associated with responses to infectious diseases such as pneumonia. In this study, we searched for single nucleotide polymorphisms (SNPs) in the coding sequences of porcine TLR1, TLR2, TLR4, TLR5, and TLR6 genes in 96 pigs from 11 breeds and elucidated 21, 11, 7, 13, and 11 SNPs, respectively, which caused amino acid substitutions in the respective TLRs. Distribution of these nonsynonymous SNPs was biased; many were located in the leucine-rich repeats, particularly in TLR1. These data demonstrated that the heterogeneity of TLR genes was preserved in various porcine breeds despite intensive breeding that was carried out for livestock improvement. It suggests that the heterogeneity in TLR genes is advantageous in increasing the possibility of survival in porcine populations.Electronic SupplementaryMaterial Supplementary material is available for this article at     

人Toll样受体9基因启动子区的生物信息学分析

目的:探讨人Toll样受体9(TLR9)基因启动子区序列特征。方法:利用生物信息学技术预测人TLR9基因启动子区域、转录因子结合位点和CpG岛分布。结果:人TLR9基因启动子区有1434个转录因子结合位点,人和小鼠保守区域内存在23个共同的转录因子结合位点,人TLR9基因启动子区包含长572 bp的CpG岛。结论:人TLR9基因启动子区相关生物信息学的研究,提高了针对启动子的研究效率,并为预测基因启动子的功能提供了重要信息。     

Toll样受体5基因多态性与中国人群脓毒症的相关性研究

目的探讨Toll样受体5（Toll-likereceptor5,TLR5）基因多态性位点与脓毒症发生风险及疾病严重程度的相关性。方法采用病例一对照研究设计,募集了255例脓毒症患者和260例对照个体。应用贝克曼公司的商用SNPstream分型技术和PCR—RFLP方法对TLR5基因的3个编码区多态性位点进行分型。采用Logistic回归分析,校正性别、年龄、吸烟和饮酒、慢性病状态、APACHEⅡ评分和脓毒症病因等混杂因素的影响,评价多态性位点与脓毒症的发生风险,以及脓毒症性休克、死亡和器官功能障碍等表型的遗传相关性。结果TLR5基因的3个多态性位点在病例和对照组中的基因型分布均呈哈．温平衡状态。这3个编码区的多态性位点与脓毒症的发生风险和疾病严重程度均无遗传学关联。结论TLR5基因的多态性位点可能在脓毒症的发生、发展和病程转归中不发挥重要作用。     

Characterization of equine and other vertebrate TLR3, TLR7, and TLR8 genes

Toll-like receptors 3, 7, and 8 (TLR3, TLR7, and TLR8) were studied in the genomes of the domestic horse and several other mammals. The messenger RNA sequences and exon/intron structures of these TLR genes were determined. An equine bacterial artificial chromosome clone containing the TLR3 gene was assigned by fluorescent in situ hybridization to the horse chromosomal location ECA27q16–q17 and this map location was confirmed using an equine radiation hybrid panel. Direct sequencing revealed 13 single-nucleotide polymorphisms in the coding regions of the equine TLR 3, 7, and 8 genes. Of these polymorphisms, 12 were not previously reported. The allelic frequency was estimated for each single-nucleotide polymorphism from genotyping data obtained for 154 animals from five horse breeds. Some of these frequencies varied significantly among different horse breeds. Domain architecture predictions for the three equine TLR protein sequences revealed several conserved regions within the variable leucine-rich repeats between the corresponding horse and cattle TLR proteins. A phylogenetic analysis did not indicate that any significant exchanges had occurred between paralogous TLR7 and TLR8 genes in 20 vertebrate species analyzed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

大规模单核苷酸多态性分析与肿瘤易感性

单核苷酸多态性(SNPs)是人类基因组中最常见的变异形式。作为第三代遗传标记,SNP在基因定位、克隆、遗传多态性方面具有广泛应用,特别是作为基因诊断标记在预防医学中具有十分重要的作用。近年来,随着人类基因组计划的发展,数以百万计的SNP被陆续发现,并可在公共数据库中免费获得。SNP数量的快速增加和SNP检测方法的发展,为其在肿瘤易感性领城的应用提供了可能。在本综述中,我们介绍了几种高通量检测SNP的分析方法,总结了大规模SNP分析技术在肿瘤易感性中的应用,介绍了目前人们对于不同人群中的SNP分析、肿瘤易感基因、个体肿瘤易感性的理解,以及研究SNP标记与肿瘤易感性关系时存在的难点。     

DNA-PKcs, but not TLR9, is required for activation of Akt by CpG-DNA

CpG-DNA and its related synthetic CpG oligodeoxynucleotides (CpG-ODNs) play an important role in immune cell survival. It has been suggested that Akt is one of the CpG-DNA-responsive serine/threonine kinases; however, the target protein of CpG-DNA that leads to Akt activation has not been elucidated. Here, we report that ex vivo stimulation of bone marrow-derived macrophages (BMDMs) from mice lacking the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) results in defective phosphorylation and activation of Akt by CpG-DNA. Unexpectedly, loss of the Toll-like receptor 9 has a minimal effect on Akt activation in response to CpG-DNA. Further in vitro analysis using purified DNA-PK and recombinant Akt proteins reveals that DNA-PK directly induces phosphorylation and activation of Akt. In addition, in BMDMs, DNA-PKcs associates with Akt upon CpG-DNA stimulation and triggers transient nuclear translocation of Akt. Thus, our findings establish a novel role for DNA-PKcs in CpG-DNA signaling and define a CpG-DNA/DNA-PKcs/Akt pathway.     

Divergent Toll-like receptors in catfish (Ictalurus punctatus): TLR5S, TLR20, TLR21

Toll-like receptors (TLR) mediate pathogen recognition in vertebrate species through detection of conserved microbial ligands. Families of TLR molecules have been described from the genomes of the teleost fish model species zebrafish and Takifugu, but much research remains to characterize the full length sequences and pathogen specificities of individual TLR members in fish. While the majority of these pathogen receptors are conserved among vertebrate species with clear orthologues present in fish for most mammalian TLRs, several interesting differences are present in the TLR repertoire of teleost fish when compared to that of mammals. A soluble form of TLR5 has been reported from salmonid fish and Takifugu rubripes which is not present in mammals, and a large group of TLRs (arbitrarily numbered 19-23) was identified from teleost genomes with no easily discernible orthologues in mammals. To better understand these teleost adaptations to the TLR family, we have isolated, sequenced, and characterized the full-length cDNA and gene sequences of TLR5S, TLR20, and TLR21 from catfish as well as studied their expression pattern in tissues. We also mapped these genes to bacterial artificial chromosome (BAC) clones for genome analysis. While TLR5S appeared to be common in teleost fish, and TLR21 is common to birds, amphibians and fish, TLR20 has only been identified in zebrafish and catfish. Phylogenetic analysis of catfish TLR20 indicated that it is closely related to murine TLR11 and TLR12, two divergent TLRs about which little is known. All three genes appear to exist in catfish as single copy genes.     

TLR9 agonists induced cell death in Burkitt's lymphoma cells is variable and influenced by TLR9 polymorphism

Toll-like receptor 9 (TLR9) triggering is a promising novel strategy to combat cancer as it induces innate and adaptive immunity responses. B-cell lymphoma is unique in this context as tumor cells express TLR9 and may harbor latent Epstein-Barr virus (EBV), a gamma-herpesvirus with remarkable oncogenic potential when latent. Latent EBV may be promoted by TLR9 triggering via suppression of lytic EBV. Here, we elaborated an initial assessment of the impact of TLR9 triggering on EBV-positive and EBV-negative B-cell lymphoma using Burkitt''s lymphoma (BL) cell lines as an in vitro model. We show that, independent of the presence of EBV, the TLR9 ligand oligodeoxynucleotide (ODN) CpG-2006 may or may not induce caspase-dependent cell death in BL cells. Moreover, ODN CpG-2006-induced cell death responses of BL cells were associated with TLR9 single-nucleotide polymorphisms (SNPs) rs5743836 or rs352140, which we detected in primary BL tumors and in peripheral blood from healthy individuals at similar frequencies. Thus, our findings suggest that the effect of TLR9 agonists on BL cells should be tested in vitro before installment of therapy and TLR9 SNPs in BL patients should be determined as potential biological markers for the therapeutic response to treatment targeting innate immunity.     

Modulation of cytokine production and enhancement of cell viability by TLR7 and TLR9 ligands during anthrax infection of macrophages

Inhalation of Bacillus anthracis, a bioterrorism agent, results in a high mortality rate despite appropriate antibiotic therapy. Macrophages appear to be a key factor in B. anthracis pathogenesis. The burst of pro-inflammatory cytokines from macrophages could be a major cause of death in anthrax. However, preactivation of Toll-like receptors (TLRs) could modify the host response. TLR ligands stimulate the release of activating cytokines but may also down-modulate the subsequent deleterious cytokine response to pathogens. We developed a cell culture model to measure macrophage responses to B. anthracis spores and bacilli. We found that germination from spores to bacilli produced a substantial stimulus for the secretion of the cytokines IL-6, TNF-alpha, IL-10, and IL-12 p40. Our studies showed that pretreatment of mouse macrophages with the TLR9 ligand ISS-1018, or the TLR7 ligands R-848 and IT-37, results in a substantial decrease in the subsequent secretion of IL-6 and TNF-alpha in response to B. anthracis infection of macrophages. Furthermore, the TLR7 and TLR9 ligands significantly decreased anthrax-induced cytotoxicity in the macrophages. These findings suggest that TLR ligands may contribute to the enhancement of innate immunity in B. anthracis infection by suppressing potentially deleterious pro-inflammatory cytokine responses and by improving macrophage viability.