宫颈上皮内瘤样病变患者高危型HPV感染基因分型分析

目的了解宫颈上皮内瘤变患者的高危型HPV的感染及其分型和不同程度宫颈病变的主要感染型别情况。方法应用型特异PCR检测宫颈癌前病变的患者的主要高危型HPV-16、18、33、58的感染及其分型情况的相关性研究。结果在本研究宫颈上皮内瘤变患者98例中,4种高危型HPV的总阳性例数为73例,HPV总的感染率为74.5%,存在多重感染。其中HPV-16、18、33、58的总感染率分别为53.1%、38.7%、17.3%和30.6%。CIN的Ⅰ/Ⅱ/Ⅲ3组患者的4种高危型HPV的感染率分别为42.9%、61.1%和93.2%。结论主要高危型HPV在宫颈上皮内瘤变患者中感染的主要型别依次为HPV16、HPV18、HPV58、HPV33,主要为HPV16和HPV18型;不同程度CIN的高危型HPV的总感染率不同,随病变程度的加重而增加。     

宫颈癌及其前期病变HPV感染与基因型分布

目的了解宫颈癌及宫颈上皮内瘤变CIN患者的HPV的感染和其基因分型及主要感染型别情况。方法应用型特异PCR检测宫颈癌及其前病变的患者的HPV感染及其主要基因分型情况的分析。结果在本研究宫颈癌及宫颈上皮内瘤变患者中,宫颈癌的HPV感染率为91．0％,CINⅠ／Ⅱ／Ⅲ的HPV感染率为73．3％,主要高危型HPV基因型别依次为HPV16、HPV18、HPV58、HPV33。结论在宫颈上皮内瘤变患者中感染主要高危型HPV基因型别依次为HPV16、HPV18、HPV58、HPV33、HPV16在宫颈癌和CIN中的构成比随着宫颈病变的增加而明显增加。     

P16 and Ki-67 expression improves the diagnostic accuracy of cervical lesions but not predict persistent high risk human papillomavirus infection with CIN1

Objective: To determine the association between the expression of p16 and Ki-67 and cervical lesions, and to evaluate the role of p16 and Ki-67 as prognostic markers for persistent high risk human papillomavirus (hr-HPV) infection. Methods: Totally 1,154 cases of cervical biopsies were enrolled, 331 cases with negative for dysplasia (NEG), 462 with cervical intraepithelial neoplasia 1 (CIN1), 176 with CIN2, 163 with CIN3 and 22 with cervical squamous cell carcinoma (SCC). Furthermore, 283 women with CIN1 were recruited into 12-month follow-up, and HPV specific gene detection by polymerase chain reaction was used to detect hr-HPV of cervical secretions at 6-month-interval for 12-month follow-up period. 40 women were infected with persistent hr-HPV, 182 with transient infection and 61 unfected with hr-HPV. The expression of p16 and Ki-67 were evaluated by immunohistochemical method. The immunostaining results of p16 and Ki-67 were classified into four categories: negative, 1+, 2+ and 3+. Results: There was significant increase in the expression of p16 (P < 0.001) and Ki-67 (P < 0.001) from NEG to SCC. The expression of Ki-67 (P < 0.001) but not p16 (P = 0.254) significantly increased in CIN2, CIN3. Ratio of p16 (P = 0.215) and Ki-67 (P = 0.495) positivity were not correlated with persistent hr-HPV infection. Conclusion: P16 and Ki-67 can improve the diagnostic accuracy of cervical lesions but can not predict persistent hr-HPV infection with CIN1.     

HPV感染与宫颈癌前病变

宫颈癌是一个由癌前病变逐渐衍变为癌的连续的病理过程。目前认为宫颈癌前病变,即宫颈上皮内瘤变与HPV感染有关。HPVs是一种双链小DNA病毒,由病毒蛋白外壳和核心DNA物质构成。病毒基因组分为早期基因区、晚期基因区及长调控区;其中早期区编码的E6、E7蛋白对于病毒的复制起关键作用。生殖道HPVs在有性活动的人群中普遍存在,在有性生活的女性中,至少有75%将在人生中的某个时间感染HPV。感染HPV后绝大多数人可以自然消退。只有感染了高危亚型、同时又具备其他高危因素的妇女才可能进展为HSIL或宫颈癌。因此对于HPV的感染既要重视,又不必恐惧。目前的治疗主要是针对由HPV引起的宫颈或外生殖器的局部病变。     

The prevalence of VAIN,CIN, and related HPV genotypes in Japanese women with abnormal cytology

Vaginal intraepithelial neoplasia (VAIN) is often found by chance. We investigated the prevalence of VAIN and related human papillomavirus (HPV) types in comparison with cervical intraepithelial neoplasia (CIN). This study enrolled 648 women who were referred to the outpatient clinic of Kanazawa Medical University Hospital for abnormal cytology from January 2009 to January 2019. HPV genotypes were determined using Genosearch-31 + 4, which can detect 35 different HPV types. Colposcopy was performed at the first visit by an experienced gynecological oncologist. Among 611 subjects with squamous cell lesions, 107 (17.5%) VAIN cases were identified, and 67 (11.0%) women had both VAIN and CIN. Ultimately, 72 VAIN1, 15 VAIN2/3, 203 CIN1, 249 CIN2/3, 32 cervical squamous cell carcinomas (SCC), and one vaginal SCC (Vag-SCC) were identified. The prevalences of VAIN1, VAIN2/3, and Vag-SCC were 35.5%, 6.0%, and 3.1% of equivalent cervical lesions, respectively. The VAIN patients were older than the CIN patients (P = .002). About half of the VAIN cases were diagnosed during the follow-up. Multiple HPV infections were found in 42.9% of the VAIN and CIN patients. HPV52, 16, 51, 53, and 56 were the most common types in VAIN, whereas HPV16, 52, 58, 51, and 31 predominated in CIN. HPV18 was rare in VAIN, HPV58 was more common in CIN than in VAIN, and HPV53 and HPV73 were more common in VAIN. In conclusion, VAIN1 was identified more frequently than we expected. Various HPV types were identified in the vagina, which is likely a reservoir for HPV.     

Association between high risk human papillomavirus infection and co-infection with Candida spp. and Trichomonas vaginalis in women with cervical premalignant and malignant lesions

BackgroundHuman papillomavirus (HPV) is the necessary cause of cervical cancer. Cervico-vaginal infection with pathogens like Chlamydia is a likely cofactor. The interactions between HPV, Trichomonas vaginalis (TV) and Candida spp. are less understood, though inflammation induced by these pathogens has been demonstrated to facilitate oncogenesis.ObjectiveOur study aimed to evaluate the association between Candida spp. and TV co-infection with HPV in cervical oncogenesis.Study designWomen with normal cervix who were high-risk HPV-negative (N = 104) and HPV-positive (N = 105); women with CIN 1 (N = 106) and CIN 2/CIN 3 (N = 62) were recruited from a community based cervical cancer screening program. Cervical cancer patients (N = 106) were recruited from a tertiary care oncology clinic. High-risk HPV was detected by Hybrid Capture II technique; Candida spp. and TV were detected by culturing the high vaginal swabs followed by microscopic examination in all. The disease status was established by histopathology in all the women.ResultHPV-positive women had significantly higher risk of having precursor lesions (of any grade) and cancer compared to HPV-negative women. Candida spp. or TV infection did not alter the risk of low grade or high grade lesions among HPV- positive women. HPV positive women co-infected with TV had higher risk of cervical cancer but not those co-infected with Candida spp.ConclusionThe higher risk of cancer observed in the women co-infected with HPV and TV without any enhanced risk of CIN 3 suggests secondary infection of the malignant growth by TV rather than any causal role. Co-infection with Candida spp. and/or TV infection did not increase the carcinogenic effect of HPV on cervix.     

CIN患者高危型HPV感染及宫颈环形电切术对其的影响

目的:分析子宫颈上皮内瘤样变(CIN)与高危型人乳头瘤病毒(HR-HPV)的关系,观察宫颈环形电切术(Leep)对HR-HPV感染的治疗作用。方法:将我院宫颈病-宫颈癌筛查防治中心同时作病理活体组织检查和HR-HPVDNA检测的893例患者,依据病理活检结果分为正常组、CIN-I组、CIN-II组、CIN-III组、SCC组;对其中HR-HPVDNA阳性的CIN-I～CINIII的92例患者,在Leep后3月复查HR-HPVDNA。结果:各组HR-HPVDNA阳性率:正常组32.18%(223/693)、CIN-I组53.66%(66/123)、CIN-II组78.72%(37/47)、CIN-III组93.10%(27/29)、SCC组100%(1/1);92例CIN患者Leep后HR-HPVDNA转阴率分别是:CIN-I组92.86%(26/28)、CIN-II组94.59%(35/37)、CIN-III组62.96%(17/27)。结论:HR-HPV感染与CIN的发生有关;Leep能有效治疗HR-HPV感染。     

The human papillomavirus (HPV) vaccine,HPV related diseases and cervical cancer in the post-reproductive years

Prophylactic HPV vaccines have demonstrated high efficacy against a range of HPV related diseases. They have been widely adopted as population health interventions in many jurisdictions and their routine use has been endorsed by the WHO. The development of these vaccines comes after an increased understanding of the natural history and epidemiology of HPV infection and disease in both males and females. Persistent HPV infection with oncogenic types induces malignant transformation in a range of epithelia including the cervix, anogenital regions, the penis and a number of head and neck sites. In relation to HPV disease prevention in the post-reproductive years, most infections occur soon after commencement of sexual activity but new infections do occur throughout the age spectrum. This reduces the likely impact of prophylactic vaccines in this population. The major impact on HPV related disease in this age group will come from advances in screening and early detection of HPV and neoplastic precursors. The most appropriate prevention for any individual man or women in this age group will be an individualised combination of vaccination, screening and early detection depending on the individual's own circumstances.     

应用表面等离子共振生物传感器分型检测HPV及其应用评价

目的评价表面等离子共振(SPR)生物传感器分型检测女性生殖道人类乳头瘤病毒(HPV)的临床应用价值。方法采集女性宫颈脱落细胞504例,按病理学结果分为炎症组、宫颈上皮内瘤变(CIN)Ⅰ组、CINⅠ-Ⅱ组、CINⅡ组、CINⅢ组、宫颈癌组。应用SPR生物传感器对各组进行检测,一次性分型检测16种HPV高危型和8种低危型,同时采用克隆测序作平行对照,结合病理诊断结果,对SPR生物传感器进行应用评价。结果SPR生物传感器和克隆测序结果的一致率为0.994,Kappa指数为0.987,(P=0.0000.05)。SPR生物传感器测得各病理组别HPV阳性率、高危型阳性率及多重感染率依次为总体(64.7%、62.1%、15.5%),炎症(41.9%、36.6%、11.8%),CINⅠ(44.6%、41.2%、13.5%),CINⅠ-Ⅱ(51.9%、48.1%、11.1%),CINⅡ(74.2%、74.2%、9.7%),CINⅢ(94.3%、93.5%、23.6%)及宫颈癌组(98.2%、98.2%、16.4%)。24种HPV基因型检出21种,阳性率依次为:16、58、33、52、66、11、18、53、6、31、45、39、81、59、70、68、51、54、56、35、40。SPR生物传感器检测HPV DNA诊断CIN III和宫颈癌的灵敏度为95.5%、特异度为52.1%、阳性预测值为52.1%、阴性预测值为95.5%。结论随宫颈病变严重程度的增高,HPV感染率和高危型感染率呈升高趋势,多重感染率无明显升高趋势;SPR生物传感器检测HPV DNA与克隆测序一致性良好,可以实现HPV分型检测,其诊断CINⅢ和宫颈癌具有较高的灵敏度和阴性预测值,在宫颈病变的临床诊断和流行病学调查中具有重要意义。     

宫颈鳞状细胞癌和癌前病变中Skp2表达及其与HPV16/18感染的关系

目的 探讨skp2在宫颈鳞状细胞癌和癌前病变中的表达规律及其与人乳头状瘤病毒(HPV)感染之间的关系.方法 采用免疫组织化学(ABC法)和原位杂交检测Skp2蛋白和HPV16/18 DNA在30例正常宫颈鳞状上皮、29例宫颈低级别上皮内瘤变、31例高级别上皮内瘤变和31例宫颈鳞状细胞癌中的表达.结果 Skp2在正常宫颈鳞状上皮中呈阴性,与宫颈低级别上皮内瘤变(阳性表达率为13.8%,4/29)之间差异无统计学意义(P>0.05).随着上皮病变级别升高,表达也逐渐增强,在宫颈鳞状细胞癌中表达更强;HPV16/18 DNA在四组中的阳性表达率,除高级别上皮内瘤变和宫颈鳞状细胞癌两组间差异无统计学意义外(均为96.8%),其余各组之间差异均有统计学意义(P<0.01);在宫颈低级别上皮内瘤变中skp2蛋白表达和HPV感染相关无统计学意义,但在高级别上皮内瘤变和宫颈鳞状细胞癌两组中均呈正相关(γ高级别=0.373,γ癌 =0.416,P<0.05).结论 Skp2过表达主要在宫颈鳞状细胞癌形成的中晚期起作用,可作为一个早期诊断恶性的指标,且可能与HPV16/18感染有协同作用.E7-skp2-Rb可能是HPV感染诱导宫颈鳞状细胞癌形成的一条新致癌途径.     

HPV mRNA检测在宫颈癌诊断中的研究

根据国际癌症研究机构(IARC)报道,仅在2012年全世界就有超过25万女性死于宫颈癌,这是女性相关癌症的第四大死因[1].不管是生物学研究还是流行病学研究都已经证实宫颈癌与人乳头瘤病毒(HPV)感染息息相关[2]l.单独的HPV感染可能会引起大约5％的病毒相关的癌症发生,但是却是宫颈癌发病的全部因素[3].目前已经有120多种不同基因型的HPV,根据其临床表现可分为低风险型和高风险型,然而只有少数的HPV能够导致癌症的发生[4].本文就HPV致宫颈癌的发生和HPV的实验室检查以及HPV mRNA的相关进展做一些简要论述.     

Fluorescence microscopy of H&E stained cervical biopsies to assist the diagnosis and grading of CIN

Background

Prevention of cervical cancer is based upon the accurate diagnosis and grading of cervical lesions identified during screening. The pathological classification of cervical intraepithelial neoplasia (CIN) is problematic, as it relies on subjective criteria and is known to have high interobserver variability and low reproducibility. These limitations can result in either over or under treatment of patients. Biomarkers to improve CIN diagnosis have not overcome all these challenges.

Significance of HPV 16 and 18 viral load quantitation in women referred for colposcopy

The clinical utility of HPV 16 and 18 viral loads remains debated. The aim of this study was to assess the clinical significance of HPV 16 and 18 viral load and to determine a cut‐off for optimal prediction of grade 2 or higher cervical intraepithelial neoplasia among patients referred to colposcopy. A total of 186 cervico‐vaginal specimens harboring HPV 16 and/or 18 obtained at the time of colposcopy from patients without previous cervical neoplasia were tested for HPV 16 and 18 detection and quantitation using quantitative duplex real‐time PCR method. Grade 2 or higher cervical intraepithelial neoplasia was diagnosed in 87 (46.8%) cases. Only HPV 16 median viral load increased significantly with the lesion grade: 9.1 × 10⁴ in normal cervix or grade 1 cervical intraepithelial lesion versus 4.0 × 10⁶ copies per million cells in grade 2 or higher cervical intraepithelial lesion (P < 0.001). The highest predictive value for grade 2 or higher cervical intraepithelial lesion was observed with a HPV 16 viral load cut‐off of 3.0 × 10⁶ copies per million cells (91% specificity, 58.2% sensitivity). Using this cut‐off, the highest predictive value of HPV 16 viral load was observed among those referred for previous low‐grade abnormal cervical cytology (96.4% specificity, 88% sensitivity). HPV 18 quantitation showed very poor predictive value. Specific attention should be given when performing colposcopic examination of women with an HPV 16 viral load higher than 3.0 × 10⁶ copies per million cells, especially among those referred after a low‐grade abnormal cytology. J. Med. Virol. 84:306–313, 2012. © 2011 Wiley Periodicals, Inc.     

UK population based study to predict impact of HPV vaccination

BackgroundIn 2008 a human papillomavirus (HPV) vaccination programme for cervical cancer prevention was implemented in the UK. Surveillance of vaccine uptake, impact on prevalence of HPV infection and cervical cancer incidence were identified as key measures to evaluate the intervention.ObjectiveTo determine baseline HPV prevalence in unvaccinated women and predict impact of HPV vaccination on high-grade cervical disease (CIN2+).Study designA pseudo-anonymous prospective cohort was sampled on entry to the routine cervical screening programme between March 2009 and November 2010. In total, 13,306 eligible females were identified and high-risk (hrHPV) type specific status determined. Potential impact of prophylactic vaccination on CIN2+ was calculated by applying HPV vaccine clinical trial data to the baseline HPV type-specific data.ResultsOf 13,306 samples tested, 3545 (26.6%) were confirmed positive for at least one hrHPV type and 1325 (10%) were positive for low risk HPV. HPV16 was the predominant type detected in cases positive with either single or multiple hrHPV infection(s) (5.2% and 4.7%, respectively). Based on hrHPV type-specific data, Gardasil would have prevented 33.2% HPV16/18 unrelated CIN2+ compared to 47.1% for Cervarix. This difference was not statistically significant.ConclusionPrior to the introduction of the HPV vaccine, approximately one-quarter of young women were positive for hrHPV and one-tenth positive for HPV16. Post-vaccination, we anticipate a substantial absolute risk reduction in high-grade cervical disease associated with both targeted and non-targeted hrHPV types. There is no significant difference between the two commercially available vaccines in terms of clinical impact.     

Sourcing of the WHO human papillomavirus type 18 international standards for HPV antibody levels

BackgroundHPV serology is important for studies of vaccine immunogenicity, but can not be performed in a comparable manner without international standardisation.ObjectivesTo find suitable candidate sera from naturally infected persons for use as International Standards (IS) for antibodies to high-risk HPVs, with priority for HPV-18.Study design946 healthy Thai women (median age 44, range 18–83) and 61 cervical cancer patients were screened using an HPV pseudovirion-Luminex assay to detect antibodies to genital (HPV-6,-11,-16,-18,-31,-33,-45,-52,-58,-68) and non-genital HPV types (HPV-5,-15,-32,-38 and -76). Suitable candidate sera should ideally be mono-specific (have reactivity against only one genital HPV) and have high antibody levels that are stable over time.ResultsSeroprevalences of HPV-16,-31,-52 and -58 were at least twice as high among cancer patients compared to healthy individuals. Thirteen healthy women who met the IS inclusion criteria in initial testing also consented to blood-bag donations. Donations from 2 women with high HPV-18 Ab titers were pooled to the HPV-18 candidate IS, later established as the WHO official IS for HPV antibodies. Sera that could potentially be used as candidate IS for other oncogenic HPVs have also been identified.ConclusionsIn the Thai population, seroepidemiology implicated HPV types HPV-16,-31,-52 and -58 as particularly associated with cervical cancer. A well characterized cohort study has allowed sourcing of materials for an IS for HPV-18 antibodies and could conceivably be used for IS for other HPV types as well.