【作者】 李晓辉；

【导师】 李其林；

【作者基本信息】 暨南大学 ， 皮肤病与性病学， 2010， 硕士

【摘要】 尖锐湿疣是由人乳头瘤病毒感染引起的一种性感染疾病,发病率高,已居性感染疾病的第二位。该病传染性强,复发率高,对人类身心危害较大。近年来研究发现,免疫失衡在尖锐湿疣的发生、发展及转归等方面发挥重要作用。Toll样受体是一类天然识别受体,在天然免疫和适应性免疫中发挥着重要的桥梁作用。Toll样受体9(Toll like receptor 9,SNP)是其中的一种,主要识别未甲基化的CpG基序,从而在抗病毒免疫和自身免疫方面发挥重要的作用。本课题着重研究了尖锐湿疣患者外周血TLR9的单核苷酸多态性(Singlenucleotide polymorphism,SNP),分析Toll样受体9的SNP与尖锐湿疣发病的相关性。研究目的探讨Toll样受体9的单核苷酸多态性与尖锐湿疣发病的相关性。研究方法抽取63例确诊的尖锐湿疣患者和23例健康对照组的外周静脉血,采用TLR9基因直接测序的方法分析TLR9受体的单核苷酸多态性。结果1.TLR9基因自翻译起始点第+1174、1635、1269、1724位均存在单核苷酸多态性位点,分别称为SNP1、SNP2、SNP3和SNP4,其中SNP3、SNP4为新发现位点,SNP1、SNP2为NCBI数据库中公布的位点,登录号分别为：rs352139、rs352140。SNP1、SNP2在人群中均可表现为三种基因型：AA纯合、AG杂合和GG纯合,这两个位点的三种基因型在尖锐湿疣患者和健康对照组间比较差异均无统计学意义(P>0.05)。SNP3、SNP4均只在1例尖锐湿疣患者中发现,均表现为AG杂合,其余个体该两位点均为GG纯合。2.尖锐湿疣患者与健康对照组的SNP1位点的等位基因A的频率分别为0.690、0.609,等位基因G的频率0.309、0.391,两组间比较差异均无统计学意义(P>0.05)。SNP2位点的等位基因A的频率分别为0.302、0.369,等位基因G的频率0.698、0.630,两组间比较差异无统计学意义(P>0.05)。3.SNP1、SNP2两个多态性位点存在4种单倍体型,分别为AA、AG、GA、GG。每种单倍体型在尖锐湿疣患者组和健康对照组间比较差异均无统计学意义(P>0.05)。4.尖锐湿疣患者初发组与复发组的CD3+分别为：63.24%±10.09%、67.11%±9.28%,两组间比较差异均无统计学意义(P>0.05)；CD4+分别为：32.13%±8.65%、30.89%±5.81%,两组间比较差异均无统计学意义(P>0.05)；CD8+分别为：29.04%±9.29%、33.00%±8.06%,两组间比较差异均无统计学意义(P>0.05)；Th／Ts分别为：1.25±0.53、1.00±0.33,两组间差异具有统计学意义(P<0.05)。5.SNP1位点AA、AG、GG三种基因型的Th／Ts分别为：1.08±0.48、0.92±0.35、1.09±0.47,三种基因型间比较差异无统计学意义(P>0.05)；SNP2位点AA、AG、GG三种基因型的Th／Ts分别为：1.08±0.42、0.92±0.37、1.08±0.48,三种基因型间比较差异无统计学意义(P>0.05)。结论1.TLR9基因在广东汉族人群中存在4个单核苷酸多态性位点,分别为SNP1、SNP2、SNP3、SNP4。2.SNP1、SNP2两位点均与尖锐湿疣的发病易感性可能无相关性。3.初发与复发的尖锐湿疣患者间存在T淋巴细胞亚群免疫失衡。4.SNP1、SNP2两位点与尖锐湿疣患者的T淋巴细胞亚群间可能无相关性。更多还原

【Abstract】 Condyloma Acuminate is a kind of Sexually Transmitted Disease (STD). In recent years, the incidence of catching this disease appears to have an upward trend, and pathogenesis of Condyloma Acuminatum is not yet thoroughly clear. There is, so far, no effective anti-viral treatment for Condyloma Acuminate, and no satisfactory solution to the recurrence. Different individuals have different responses to HPV infection, suggesting that immunity, particularly, the host cell immunity, has a certain impact in the occurrence of Condyloma Acuminate. It has been widely recognized that the change of cellular immune function closely relates to the occurrence of Condyloma Acuminate. That the body can not build an effective immune response is the main reason for HPV persistent infection; so to explore the immune mechanisms of Condyloma Acuminate has important oretical and clinical significance.Toll-like receptor (TLR) is the first recognition receptor of the inherent immunity, which is a defending response caused by direct perception of microbial pathogen. This kind of receptor plays an important part in inherent immunity defense, and can also adjust adaptive immunity, which makes it a bridge of connecting inherent immunity and adaptive immunity. TLR9 mainly identifies bacteria and virus genome DNA and synthetic CpG ODNs, and it has an important role in the pathogenesis of anti-infection, anti-tumor, mediated cell apoptosis and autoimmune disease.ObjectiveThis study aims at investigating whether polymorphism in the TLR9 gene differentially affect the incidence and the clinical course of Condyloma Acuminatum.MethodsFollowing informed consent from all patients and healthy volunteers,2ml of peripheral blood was collected from each of 63 Guangdong Condyloma Acuminatum patients and 23 unrelated healthy Guangdong individuals admitted to The Fourth Affiliated Hospital of Jinan University. The patients were divided into groups in terms of the primary and recurrent Condyloma Acuminatum patients. With DNA samples from all subjects prepared, all single nucleotide polymorphisms of TLR9 was tested through polymerase chain reaction and direct sequencing. ResultsThere are 4 SNPs in TLR9:SNP1、SNP2、SNP3 and SNP4. SNP1 was located in intron 1 region, SNP2、SNP3 and SNP4 located in exon2. The registration number of SNP1 is rs352139 in NCBI database, and that of SNP2 is rs352140. SNP2 is non-synonymous cSNP. Based on a SNP map that had been constructed, we also constructed a haplotype map of TLR9. The frequency of alleles and genotypes in chinses was revealed as well. But there were no significantly increase in patients compared with controls on SNP1 and SNP2. There are statistical differences between the two groups of the primary and recurrent Condyloma Acuminatum patients on Th/Ts. Two sites of three genotypes were not statistically significant compared with Th/Ts. SNP3 and SNP4 were observed only in the patients, therefore, we don’t have analyzed these two sites.ConclusionsTLR9 genes exist as four single nucleotide polymorphisms (SNPs), respectively, SNP2, SNP1, SNP3, SNP4, in Guangdong Han ethnic population. Compared with the control group, no significant increase of single nucleotide polymorphisms in TLR9 can be seen in the patient group. But the data of TLR9 polymorphism in Guangdong Han Ethnic Group will significantly contribute to the improvement of research on disease association and basic immunology.更多还原