细胞因子在HR-HPV持续感染宫颈局部免疫微环境中的作用

发布时间：2018-07-08 15:41

本文选题：人乳头瘤病毒 + 宫颈癌　；参考：《天津医科大学》2017年硕士论文

【摘要】：目的:研究宫颈高危型人乳头瘤病毒(high risk human papillomavirus,HR-HPV)持续感染过程中宫颈局部Th1/Th2/Th17/Treg细胞相关的8种细胞因子的变化,包括白介素-2(interleukin-2,IL-2)、IL-4、IL-6、IL-10、干扰素-γ(γ-interferon,IFN-γ)、肿瘤坏死因子(tumor necrosis factor,TNF)、IL-17A及转化生长因子-β(transforming growth factor-β,TGF-β)。进一步分析不同HR-HPV感染亚型引起的几种细胞因子浓度变化,对IL-6的单核苷酸多态性(single nucleotide polymorphism,SNP)进行检测,分析多态性对HR-HPV持续感染及IL-6浓度的影响,为预防HR-HPV持续感染及宫颈病变进展提供理论依据。方法:选取2015年11月至2016年6月期间就诊于天津市中心妇产科医院阴道镜门诊的有HR-HPV感染史的210位已婚女性,所有研究对象均需签署知情同意书。按照HR-HPV感染情况分为两组,既往HR-HPV感染转阴组及HR-HPV持续感染组各105例,持续感染组按照宫颈活检病理进一步分为无宫颈鳞状上皮内病变组(non squamous intraepithelial lesion,NSIL)35例、低度宫颈上皮内病变组(low grade squamous intraepithelial lesion,LSIL)35例、高度宫颈上皮内病变组(high grade squamous intraepithelial lesion,HSIL)35例。采集研究对象阴道灌洗液(以下简称灌洗液)及宫颈脱落细胞。用流式BD CBA(cytometric bead array)及酶联免疫吸附法(enzyme-linked immunosorbent assay,Elisa)对灌洗液中8种细胞因子水平进行检测,提取宫颈脱落细胞中的DNA,使用巢式PCR(polymerase chain reaction)及焦磷酸测序技术对HR-HPV进行分型及IL-6两个位点rs1800795(-174 G/C)、rs1800796(-572 C/G)的SNP检测。应用SPSS17.0对数据进行分析,计量资料均采用均数±标准差(??±s)表示,组间比较采用单因素方差分析,进一步的两组比较用LSD-t法;两组间比较使用独立样本t检验,计数资料采用卡方检验,P0.05为差异有统计学意义。结果:1.IL-2浓度在HR-HPV持续感染组低于转阴组,且随着病变加重浓度逐渐降低(P0.05),IL-6浓度变化恰好相反,余6种细胞因子水平在组间差异无统计学意义。2.转阴组及持续感染组中各细胞因子浓度均与绝经状态无相关性。3.HR-HPV持续感染者中最常见的型别为HPV16(50.48%)。4.与其他高危亚型相比,HPV16感染引起的宫颈局部细胞因子变化并无差异。5.IL-6 rs1800795(-174G/C)、rs1800796(-572 C/G)基因型及等位基因频率在HR-HPV持续感染组与转阴组比较无差异。6.IL-6浓度与rs1800795(-174G/C)、rs1800796(-572 C/G)多态性无相关性。结论:1.用流式BD CBA方法进行阴道局部灌洗液中细胞因子的检测为研究HR-HPV持续感染过程中宫颈局部免疫微环境改变提供了新的思路。2.IL-2、IL-6在HR-HPV持续感染过程中有重要作用,IL-2随病变程度加重浓度逐渐降低,IL-6随病变加重浓度逐渐升高,HR-HPV持续感染过程中存在Th1/Th2漂移现象。3.HR-HPV持续感染最常见的亚型为HPV16。但HPV16与其他高危亚型感染相比引起宫颈局部8种细胞因子水平的改变无差异。4.IL-6 SNP的研究为从遗传学角度分析HR-HPV持续感染的原因提供了思路,发现影响HR-HPV持续感染的多态性位点会为预防HR-HPV持续感染及病变进展提供新的理论依据。
[Abstract]:Objective: To study the changes of 8 cytokines related to cervical local Th1/Th2/Th17/Treg cells during the continuous infection of cervical high-risk human papillomavirus (high risk human papillomavirus, HR-HPV), including interleukin-2 (IL-2), IL-4, IL-6, IL-10, interferon gamma, tumor necrosis factor. Factor, TNF), IL-17A and transforming growth factor - beta (transforming growth factor- beta, TGF- beta). Further analysis of the changes in the concentration of several cytokines caused by different HR-HPV infection subtypes, and detect the single nucleotide polymorphisms of IL-6 (single nucleotide polymorphism), and analyze the effect of polymorphism on persistent infection and concentration of HR-HPV. To provide a theoretical basis for preventing HR-HPV persistent infection and progression of cervical lesions. Methods: 210 married women who had a history of HR-HPV infection in the colposcopy clinic of Tianjin Central Obstetrics and Gynecology Hospital from November 2015 to June 2016 were selected. All the subjects needed to sign informed consent. According to the HR-HPV infection, two groups were divided into H. The R-HPV infection group and the HR-HPV continuous infection group were 105, and the continuous infection group was further divided into 35 cases without cervical squamous intraepithelial lesion (non squamous intraepithelial lesion, NSIL), 35 cases of low cervical intraepithelial lesion (low grade squamous intraepithelial lesion), and high cervical intraepithelial disease. 35 cases of high grade squamous intraepithelial lesion (HSIL) were used to collect the vaginal lavage fluid (hereinafter referred to as the lavage solution) and the cervical exfoliative cells. The level of 8 cytokines in the lavage liquid was detected by flow BD CBA (cytometric bead array) and enzyme linked immunosorbent assay. DNA in the exfoliated cells of the cervix, using polymerase chain reaction and pyrosequencing technology to classify HR-HPV and IL-6 two loci rs1800795 (-174 G/C), rs1800796 (-572). The two groups were compared with the LSD-t method; the two groups were compared with the independent sample t test, the count data were taken by chi square test, and the difference of P0.05 was statistically significant. Results: the concentration of 1.IL-2 in HR-HPV continuous infection group was lower than that in the negative group, and with the increasing concentration of the lesions (P0.05), the IL-6 concentration changed exactly the opposite, the remaining 6 species were the opposite. There was no significant difference in the level of cytokines between the groups and the.2. negative group and the persistent infection group. The most common type of.3.HR-HPV persistent infection in the patients with menopause was the most common type of HPV16 (50.48%).4., compared with other high-risk subtypes, the changes of cervical local cytokines caused by HPV16 infection did not differ from.5.IL-6 rs. 1800795 (-174G/C), rs1800796 (-572 C/G) genotype and allele frequency had no difference between.6.IL-6 concentration and rs1800795 (-174G/C), rs1800796 (-572 C/G) polymorphism in the HR-HPV continuous infection group and the negative group. Conclusion: 1. the detection of cytokine in the local lavage liquid of the vaginal canal was carried out by the flow BD method. The local immune microenvironment changes during the dyeing process provide a new idea.2.IL-2. IL-6 plays an important role in the continuous infection process of HR-HPV. The concentration of IL-2 gradually decreases with the severity of the lesion, IL-6 increases with the aggravation of the lesion, and the most common subtype of Th1/Th2 drift in.3.HR-HPV continues to exist during the continuous HR-HPV infection. The study of changes in the level of 8 cytokine levels of the cervix caused by HPV16. but HPV16 compared with other high-risk subtypes of.4.IL-6 SNP provides a way of thinking to analyze the causes of persistent HR-HPV infection from the genetic perspective. It is found that polymorphic loci affecting the continuous infection of HR-HPV will provide the prevention of HR-HPV persistent infection and the progression of the disease. A new theoretical basis.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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