Comparison of HPV genotypes and viral load between different sites of genital tract: The significance for cervical cancer screening

ObjectivesTo compare the consistency of HPV genotype and viral loads among different sites within the female genital tract, and to correlate these with clinical outcomes.Methods2646 previously unscreened rural women were enrolled in this population-based, cross-sectional study between May 2006 and April 2007. Physician-collected samples from lower vagina, upper vagina, cervix, and one self-collected sample were taken from each woman. Viral load was assessed by HC2 using the relative light unit/cutoff ratio (RLU/CO), and HPV genotyping was tested by Linear Array.ResultsThe low risk HPV positive rate was highest in lower vagina samples and lowest in cervix samples. Overall kappa values of high risk HPV types between various anatomic sampling sites showed substantial or almost perfect agreement among women with normal pathology, CIN1, and CIN2+. In the CIN2+ population, high risk HPV viral load for cervix samples (557.25 RLU/CO) were much higher than upper vagina samples (96.43 RLU/CO, P < 0.001), lower vagina samples (36.51 RLU/CO, P < 0.001), and self-collected (206.83 RLU/CO, P = 0.003) samples.ConclusionsAlthough the distribution of high risk HPV genotypes was fairly equivalent across different genital sites, particularly for CIN2+ lesions, viral loads were largely variable. The findings may affect the cervical cancer screening methods using self-collected samples, particularly in resource-challenged areas.     

Human papillomavirus infection in women with and without cervical cancer in Tbilisi, Georgia

Background: No accurate estimates of cervical cancer incidence or mortality currently exist in Georgia. Nor are there any data on the population-based prevalence of high-risk (HR) human papillomavirus (HPV) infection, which, in the absence of good-quality screening, is known to correlate with cervical cancer incidence. Methods: We obtained cervical cell specimens from 1309 women aged 18–59 years from the general population of Tbilisi, and also from 91 locally diagnosed invasive cervical cancers (ICC). DNA of 44 HPV types was tested for using a GP5+/6+-based PCR assay. Results: In the general population (of whom 2% reported a previous Pap smear) HPV prevalence was 13.5% (95% CI: 11.6–15.9), being highest in women aged 25–34 years (18.7%) and falling to between 8.6% and 9.5% for all age groups above 34 years. HR HPV prevalence was 8.6% overall, being 6.8% and 38.9% among women with normal and abnormal cytology, respectively. HPV45 (1.6%) was the most common type in women with normal cytology, whereas HPV16 predominated among women with cervical abnormalities (including 7 of 10 histologically confirmed cervical intraepithelial neoplasia 2/3) and among ICC (57.6%). The next most common types in ICC in Georgia were HPV45 and 18 (13.2 and 11.0%, respectively). Conclusions: We report a relatively high burden of HPV infection in Tbilisi, Georgia. Improving cervical cancer prevention, through screening and/or HPV vaccination, is an important public health issue in Georgia, where 70% of ICC are theoretically preventable by HPV16/18 vaccines.     

Human papillomavirus genotype distribution in cervical intraepithelial neoplasia grades 1 or worse among 4215 Chinese women in a population-based study

PurposeTo estimate the burden of human papillomavirus (HPV) infection and cervical disease among sexually active women in a sample of Chinese women.MethodsA multicenter, population-based study was conducted between May 2006 and April 2007. A total of 4215 sexually active women aged 17–54 years were surveyed from five geographical sites representing both urban and rural areas: Beijing, Shanghai, Shanxi, Henan and Xinjiang. Women were referred for colposcopy on the basis of results of Pap testing and HPV screening. HPV genotyping of the CIN1+ specimens was performed with INNO-LiPA. Attribution of HPV types to lesions was estimated using a fractional contribution approach.Results13.3% of the women (559/4215) were referred for colposcopy; 4.3% (183/4215) of these were diagnosed with CIN1+. Of the latter, 88.5% (162/183) were typed and 94.4% (153/162) were HPV-positive. HPV16 was the most prevalent type in lesions in both urban and rural settings. Combined, HPV16 and 18 were attributable to 71.4% of HPV-positive CIN2+ lesions. In addition, HPV31, 33, 52 and 58 were prevalent in CIN1+ lesions, with HPV33, 52, and 58 combined accounting for 24.1% CIN2+ lesions. Though prevalent, HPV31 always occurred as a co-infection with another HPV type and therefore was attributed minimal causality.ConclusionsHPV16 and 18 are associated with the majority of cervical lesions in Chinese women from which this population-based sample was drawn. In addition, other HPV types, such as 33, 52, and 58, also play an important role in cervical disease.     

人乳头瘤病毒感染与宫颈癌的研究进展

人乳头瘤病毒(Human Papillomavirus HPV)感染是导致性传播疾病的常见原因,上世纪八十年代初,首次报道生殖器HPV感染与宫颈癌之间的联系,认为HPV感染是95%以上宫颈癌变的高危因素。随着分子生物学技术的发展,对HPV致癌机制的研究不断深入,取得大量有价值的成果,现就HPV的致癌途径与协同因素探讨宫颈癌的发病机制以及对HPV检测方法等方面的研究进行综述。     

一氧化氮、人乳头瘤病毒和宫颈癌之间相互关系的研究进展

人乳头瘤病毒感染是宫颈癌发生的重要始动原因,从HPV感染到宫颈癌发生,需要许多共刺激因子的参与。这些共刺激因子均可引起宫颈局部一氧化氮浓度的增高。而一氧化氮既可影响HPV的转录和翻译,又在肿瘤发生过程中具有重要调节作用。深入研究一氧化氮、人乳头瘤病毒感染及宫颈癌之间的关系,可为宫颈癌的防治提供新的重要理论基础和药物研制实验平台,通过使用一氧化氮合酶抑制剂降低宫颈局部NO浓度将为全面有效防治宫颈癌带来新的希望。     

Risk factors associated with human papillomavirus prevalence and cervical neoplasia among Cameroonian women

BackgroundThis study used community-based cervical cancer screening for high-risk human-papillomavirus (HPV) to determine demographic and lifestyle factors associated with HPV prevalence and cervical intraepithelial neoplasia grade 2 or worse (CIN2+).MethodsWomen (n = 838) aged 25–65 years were recruited in two sequential studies in Cameroon. Demographic and historical data were obtained from participants and specimens were self-collected for HPV-testing using real-time PCR. HPV-positive women underwent biopsy and endocervical curettage. Associations were determined using bivariate analysis and logistic regression.ResultsHPV and self-reported HIV prevalence were 39.0% and 9.2%, respectively. Eighteen (9.3%) CIN2+ lesions were found among HPV-positive women. Housewives had a higher risk of being HPV infected (OR = 1.60, p = 0.010). HIV co-infection (aOR = 3.44, p < 0.001) and hormonal contraception (aOR = 1.97, p = 0.007) were associated with increased HPV prevalence. HPV-positive women who used condoms during sexual intercourse were at lower risk of CIN2+ (aOR = 0.15, p = 0.029). CIN2–3 lesions were found in women younger than 50 years, with a median age of 36 years (31–44). HPV-16/18-positive women had a 4.65-fold increased risk of CIN2+ (p = 0.015).ConclusionsYoung, single women and housewives were at higher risk of HPV infection. Preventive strategies for cervical cancer in low-resource settings should target women aged 30–50 years for HPV screening, and should focus treatment and follow-up on HPV-16/18-positive women. Further studies are needed to clarify if other risk factors require attention.     

Detection of human papillomavirus types by the polymerase chain reaction and the differentiation between high-risk and low-risk cervical lesions

By means of a consensus polymerase chain reaction (PCR) method, the prevalence of HPV types was determined in cervical biopsies from 137 women referred to the gynecological outpatient clinic for colposcopy because of an abnormal cervical smear. The prevalence of HPV was 80.3%. There was a statistically highly significant rise in the prevalence of the oncogenic HPV types (16, 18, 31, 33) with increasing severity of cervical intraepithelial neoplasia (CIN I to III), indicating a role for these HPV types in the pathogenesis of cervical cancer. The prevalence of other HPV types decreased significantly with the severity of the lesion, suggesting that these HPV types play a less significant role in this process. These data indicate that HPV typing with PCR may be a valuable tool for distinguishing between highrisk and low-risk cervical lesions. Furthermore, our results suggest that the detection of HPV types by consensus PCR in the cervix of patients with an abnormal smear but without histologically detectable CIN is a useful tool for predicting which of these patiens will eventually develop CIN. Finally, a relatively low percentage (3%) of HPV double infections is reported in this study.     

阴道微生态改变与人乳头瘤病毒感染和宫颈病变相关研究

目的 探讨阴道微生态变化对人乳头瘤病毒(Human papillomavirus, HPV)感染和宫颈病变的影响。 方法 收集我院2018年11月至2019年7月门诊患者1 529例,进行阴道微生态和宫颈HPV检测,根据筛查结果阴道镜下多点活检,应用卡方检验分析阴道微生态变化在HPV阴性组和阳性组之间差异以及对宫颈病变程度的影响。 结果 pH值>4.6、白细胞计数增高、清洁度级别增高、乳杆菌消失、BV和滴虫感染能增加HPV感染(χ2值分别为12.281、17.692、79.999、192.700、48.863和16.758,均P结论 阴道微生态变化能增加HPV感染概率,但与HPV相关性宫颈病变关系不密切。     

宫颈上皮内瘤变患者阴道微生态状况与LEEP术后预后关系

目的 研究宫颈上皮内瘤变（CIN）患者阴道微生态状况及LEEP术后阴道微生态状况的变化特点,分析阴道微生态状况与CIN患者预后的关系。方法 回顾性分析2016年1月至2018年1月温州医科大学定理临床学院122例高危型人乳头瘤病毒（hr-HPV）阳性的CIN患者LEEP术治疗前后阴道微生态的变化特点。结果 LEEP术后菌群密集度Ⅱ~Ⅲ级（74.59%）、多样性Ⅱ~Ⅲ级（68.03%）、乳酸分级Ⅰ~Ⅱa级（70.49%）出现增多,需氧菌性阴道病（AV）（9.84%）、细菌性阴道病（BV）（14.75%）、外阴阴道假丝酵母菌病（VVC）（12.30%）以及滴虫性阴道炎（2.46%）的检出率降低和pH值（4.2±0.5）降低,与术前比较差异具有统计学意义（均P<0.05）;术后复诊hr-HPV阳性CIN患者菌群密集度Ⅱ~Ⅲ级（35.00%）、多样性Ⅱ~Ⅲ级（40.00%）、乳酸分级Ⅰ~Ⅱa级（25.00%）低于hr-HPV阴性CIN患者,AV（30.00%）、BV（35.00%）、VVC（30.00%）检出率,pH值（4.9±0.4）高于hr-HPV阴性CIN患者,差异有统计学意义（均P<0.05）。结论 阴道微生态失衡可降低宫颈免疫力,促进CIN进展、癌变,改善阴道微生态可抑制hr-HPV的感染,抑制宫颈细胞的异常增生、癌变。     

成都市人乳头瘤病毒16型感染与宫颈病变的相关性分析

本研究旨在探讨人乳头瘤病毒(human papillomavirus,HPV)16感染与宫颈病变的关系,为宫颈癌防治提供科学依据。通过核酸杂交法进行HPV感染分型,纳入1 057例HPV阳性且行组织切片病理学检查的患者,对各级别宫颈病变中HPV16构成比、不同年龄组HPV16阳性患者中宫颈上皮内瘤样病变(cervical intraepithelial neoplasia,CIN)Ⅱ及以上病变的患病率,以及HPV16单一与多重感染患者中CINⅡ及以上病变的患病率进行分析。结果显示,在1 057例HPV阳性患者中,352例感染HPV16,CINⅢ中HPV16构成比最高,各级别病变中HPV16构成比差异有统计学意义。随着病变级别增加,HPV16构成比有增高趋势(P0.05)。不同年龄组HPV16阳性患者中CINⅡ及以上病变的患病率差异有统计学意义(P0.05),且随年龄增加而升高(P0.05)。HPV16单一、双重与三重以上感染患者中,CINⅡ及以上病变的患病率差异有统计学意义(P0.05),且随着感染型别种类增加,患病率降低(P0.05)。本研究显示,HPV16与高级别宫颈病变有较明显的相关性,老年HPV16阳性患者检出宫颈癌的概率更高。因此,应高度重视HPV16持续性感染,做到及时诊断与治疗,以减少宫颈高级别病变和宫颈癌的发生。     

STAT3基因多态性与女性人乳头瘤病毒感染及宫颈病变的关系

目的探究女性人乳头瘤病毒(HPV)感染及转录活化因子3(STAT3)单核苷酸基因多态性(SNPs)的改变。方法2016年9月至2019年9月经病理组织学确定的健康子宫者60例(NC组)、低度鳞状上皮内病变(LSIL)50例(LSIL组)、高度鳞状上皮内病变(HSIL)55例(HSIL组)和宫颈鳞状细胞癌(CSCC)57例(CSCC组),PCR测定HPV感染情况,同时采用PCR 限制片段长度多态性(RFLP)测定各组STAT3基因C1697G多态性。结果HSIL组、LSIL组及CSCC组高危HPV与NC组比较阳性率显著升高,CSCC组(98.24%)显著高于HSIL组(72.73%)和LSIL组(5.8%),HSIL组显著高于LSIL组,结果均具有统计学差异(P<0.05)。LSIL组、HSIL组及CSCC组中与NC组比较显著升高的基因比例为C/C型,CSCC组C/C型基因比例为70.18%,HSIL组(58.18%)C/C型基因比例显著高于LSIL组(0.18%)(P<0.05)。与HPV阴性比较,C/C型基因比例在HPV阳性中显著升高(P<0.05)。通过调整年龄因素,C/G基因型HPV感染风险上升到3.120(95％CI:0.847～4.995),发生宫颈癌的风险上升到3.876(95％CI为0.935～5.336);C/C型基因HPV感染风险上升到3.574(95％CI为0.893～4.912),发生宫颈癌的风险上升到5.137(95％CI为0.967～5.776)。结论STAT3基因C1697G多态性与HPV易感性及宫颈病变的发生发展有密切关系。     

Haemochromatosis gene (HFE) mutations in viral-associated neoplasia: Linkage to cervical cancer

The present study examines the frequency of the two main HFE mutations (C282Y and H63D) in a randomly selected population of 346 individuals including 201 DNA samples from women with cervical neoplasia (including high-grade squamous intraepithelial lesions and invasive squamous cell carcinoma) and a control population of 146 women from the same geographical area. We found a significantly lower risk of development of cervical neoplasia in H63D carriers (OR = 0.56; 95% CI 0.35-0.92; p = 0.01). Multivariate logistic regression analysis confirms this observation (OR = 0.55; 95% CI 0.35-0.88, p = 0.01). Regarding the C282Y mutation no association was found (OR = 1.32; 95% CI 0.53-3.33; p = 0.52). In addition, a significant difference between H63D carrier and non-carrier women on the time-to-onset of cervical lesions was observed (log-rank test: p = 0.0012). These results indicate that HFE could be considered a candidate modifier gene of viral-related neoplasia such as cervical carcinoma possibly by a dual role on iron metabolism and immunological system.     

高危型人乳头瘤病毒感染与宫颈癌中miR-218表达的关系

目的观察高危型人乳头瘤病毒(HPV)感染与宫颈癌中miR-218表达的关系。方法收集2015年6月至2018年12月我院手术切除的宫颈癌组织并检测高危型HPV感染情况和miR-218表达量。培养HPV16阳性的SiHa细胞株并进行分组,阴性对照(NC)组转染NC模拟物、miR-218组转染miR-218模拟物,检测两组细胞凋亡率、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关x蛋白(Bax)、Bcl-2相互作用细胞死亡介导因子(Bim)、含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-9、Caspase-3的mRNA表达量及凋亡通路分子c-Jun氨基末端激酶(JNK)、c-Jun基因(c-Jun)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)的蛋白表达量。结果高危型HPV阳性的宫颈癌组织中miR-218表达量减少。转染24 h后,miR-218组细胞凋亡率、细胞中Bax、Bim、Caspase-9、Caspase-3的mRNA表达量及JNK、c-Jun的蛋白表达量均明显高于NC组,而细胞中Bcl-2的mRNA表达量及PI3K、AKT、mTOR的蛋白表达量均低于NC组,差异均有统计学意义(均P<0.05)。结论miR-218在高危型HPV感染的宫颈癌组织中表达减少。增加miR-218的表达能够促进HPV感染宫颈癌细胞的凋亡。该调控作用与JNK/c-Jun通路的激活及PI3K/AKT/mTOR通路的抑制有关。     

Human papillomavirus genotyping by multiplex pyrosequencing in cervical cancer patients from India

Cervical cancer is a leading cause of cancer-related deaths among women in India.Human papillomavirus (HPV) infection is the causative agent of cervical cancer; and infection with the high-risk genotypes, predominantly HPV16 and 18,is the biggest risk factor.Vaccines targeting HPV16 and 18 have been found to confer protection in large- scale clinical trials.HPV genotyping has traditionally been carried out to screen the population "at risk" using indirect methods based on polymerase chain reaction (PCR) using consensus primers combined with various DNA hybridization techniques,and often followed by the sequencing of candidate products.Recently,a high-throughput and direct method based on DNA sequencing has been described for HPV genotyping using multiplex pyrosequencing. We present a pilot study on HPV genotyping of cervical cancer and non-malignant cervical samples using multiplex pyrosequencing.Using genomic DNA from cell lines,cervical biopsies,surgical tissues or formalin-fixed,paraffin- embedded tissue samples,we could successfully resolve 6 different HPV types out of the 7 tested,with their prevalence found to be in agreement with earlier reports. We also resolved coinfections with two different HPV types in several samples. An HPV16 genotype with a specific and recurrent sequence variation was observed in 8 cancer samples and one non-malignant sample. We find this technique eminently suited for high-throughput applications,which can be easily extended to large sample cohorts to determine a robust benchmark for HPV genotypes prevalent in India.     

人乳头状瘤病毒分型及病毒载量与宫颈病变的研究进展

宫颈癌作为目前妇女最常见的恶性肿瘤,新发病例在全球范围内仍处于上升趋势。已有实验证明,HPV感染是宫颈癌发生发展的必要条件。然而不同分型及其病毒载量高低仍会影响诊断率,从而影响病情,使之发展为持续性感染,最终演变为癌前病变或宫颈癌。目前,研究者对于高危型HPV病毒载量与宫颈病变之间是否存在依存关系尚存争议。研究表明,高病毒载量可能成为预测宫颈癌前病变进展的指标,然而对于病理诊断明确的宫颈癌患者而言,病毒载量并不能代表病理分期。本文综述了人乳头状瘤病毒,不同基因分型,病毒载量等因素对于宫颈病变及宫颈癌的相关性的近年研究进展,希望有助于指导临床工作。     

人乳头状瘤病毒16型L1蛋白的高效表达及抗原性分析

为了在大肠杆菌中高效表达人乳头状瘤病毒16型L1蛋白,将L1基因的N端截去,选择最适合L1表达的载体,并且对表达条件进行筛选,结果表明重组蛋白的表达量达到33%;经包涵体变性复性和初步纯化,得到初纯的蛋白,ELISA结果分析表明,复性的L1蛋白保留了较好的抗原性。     

Changes in type‐specific human papillomavirus load predict progression to cervical cancer

Persistent high-risk human papillomavirus (HPV) infection is strongly associated with the development of high-grade cervical intraepithelial neoplasia or cancer (CIN3+). However, HPV infection is common and usually transient. Viral load measured at a single time-point is a poor predictor of the natural history of HPV infection. The profile of viral load evolution over time could distinguish HPV infections with carcinogenic potential from infections that regress. A case-cohort natural history study was set-up using a Belgian laboratory database processing more than 100,000 liquid cytology specimens annually. All cytology leftovers were submitted to real-time PCR testing identifying E6/E7 genes of 17 HPV types, with viral load expressed as HPV copies/cell. Samples from untreated women who developed CIN3+ (n = 138) and women with transient HPV infection (n = 601) who contributed at least three viral load measurements were studied. Only single-type HPV infections were selected. The changes in viral load over time were assessed by the linear regression slope for the productive and/or clearing phase of infection in women developing CIN3+ and women with transient infection respectively. Transient HPV infections generated similar increasing (0.21 copies/cell/day) and decreasing (−0.28 copies/cell/day) viral load slopes. In HPV infections leading to CIN3+, the viral load increased almost linearly with a slope of 0.0028 copies/cell/day. Difference in slopes between transient infections and infections leading to CIN3+ was highly significant (P < .0001). Serial type-specific viral load measurements predict the natural history of HPV infections and could be used to triage women in HPV-based cervical cancer screening.     

CIN患者阴道微生态状况与HPV感染的关系分析

分析宫颈上皮内瘤变（CIN）患者阴道微生态与人乳头瘤病毒（HPV）感染的关系。

选取326例本院收治的CIN患者,采集阴道分泌物和宫颈脱落细胞,通过形态学和功能学检测阴道微生态状况,采用PCR-反向点杂交法进行HPV分型检测,采用二分类Logistic回归分析影响CIN患者HPV感染的因素。

CIN患者HPV阳性共256例,占78.5%,HPV阴性共70例,占21.5%。HPV阳性CIN患者白细胞计数＞10个/高倍视野、阴道清洁度Ⅲ-Ⅳ度、VVC阳性、AV阳性、BV阳性、pH值＞4.5、白细胞酯酶阳性、过氧化氢阳性占比高于HPV阴性患者,微生态正常者占比低于HPV阴性患者（P＜0.05）。二分类Logistic回归分析显示,VVC、BV、微生态正常和pH值为CIN患者HPV感染的影响因素。

CIN患者HPV感染率较高,且感染后存在阴道微生态失调,VVC、BV、微生态异常和pH值是CIN患者HPV感染的影响因素。

     

宫颈癌患者人乳头瘤病毒感染情况及STING、ZEB1蛋白水平分析

分析宫颈癌患者人乳头瘤病毒（HPV）感染情况及癌灶组织内干扰素基因刺激因子（STING）、E盒锌指结合蛋白1（ZEB1）表达情况,为该类患者的治疗提供参考。

收集我院病理检验科2019年1月至2022年1月63例宫颈癌标本（宫颈癌组）、55例宫颈上皮内瘤变（CIN）标本（CIN组）及50例正常宫颈标本（正常组）,采用免疫组化法检测各标本内STING及ZEB1表达情况,采用原位杂交法检测各标本内高危HPV感染情况,分析STING及ZEB1蛋白表达情况与宫颈癌病理特征及高危HPV感染之间的关系。

宫颈癌组标本STING及ZEB1表达量均高于CIN组及正常组,CIN组标本ZEB1表达量高于正常组,差异均有统计学意义（均P＜0.05）;CIN组标本STING表达量与正常组比较差异无统计学意义（P＞0.05）。宫颈癌组标本HPV16、18检出率高于CIN组及正常组,CIN组标本HPV16、18检出率高于正常组,差异均有统计学意义（均P＜0.05）。宫颈癌患者FIGO分期及浸润深度与其组织内STING及ZEB1蛋白阳性情况均存在相关性;此外,宫颈癌淋巴结转移及肿瘤分化程度与组织内STING蛋白阳性情况存在相关性,差异均有统计学意义（均P＜0.05）。宫颈癌患者STING及ZEB1蛋白表达与其HPV16、18检出率间均呈正相关（均P＜0.05）。

宫颈癌组标本STING及ZEB1蛋白表达量均高于正常组及CIN组,且其表达量还与宫颈癌浸润、FIGO分期相关。STING和ZEB1可能与HPV16、18感染共同参与宫颈癌的发生及进展。