人乳头瘤病毒感染与人乳头瘤病毒疫苗的研究进展

人乳头瘤病毒(HPV)感染与宫颈癌的发生密切相关,是宫颈癌发生的最主要诱发因素.预防性HPV疫苗是一种预防宫颈癌的新方法,其效果得到了多项临床试验的肯定.治疗性HPV疫苗的研发同样备受关注,目前治疗性疫苗的类型很多,但因其机制较复杂,大多仍处在实验阶段.     

Induction of immune responses against human papillomaviruses by hypervariable epitope constructs

An ideal prophylactic vaccine against human papillomaviruses (HPV) would be one that can induce broadly reactive antibody titres to at least the major oncogenic strains of HPV. It has been previously shown that HPV structural proteins are highly immunogenic but fail to elicit cross-reactive immune responses against heterologous strains of HPV. Recent studies have demonstrated that the immunity induced by virus-like particles is mostly type specific. In the present study, we determined the breadth of reactivity of antibodies induced in mice immunized with hypervariable epitope constructs (HECs), which represent sequence variants of immunodominant B-cell epitopes of the major capsid protein L1 of HPV. In order to test the breadth of reactivity, sera from immunized mice were tested against peptides representing analogous sequences of HPV types 16, 18, 31 and 45. Mice immunized with HECs based on two epitopes mounted antibody responses that cross-reacted with two different analogues, 16 and 18. Significantly, antibodies from mice immunized with HECs also inhibited haemagglutination mediated by HPV-16 L1 VLPs, suggesting that immunization resulted in the development of antibodies that could bind to viral capsid proteins in their native conformation. Our observations suggest that HECs may overcome the restriction of type specific immunity against HPV.     

Commercially available molecular tests for human papillomaviruses: a global overview

BackgroundMolecular tests for detection of human papillomaviruses (HPVs) play a crucial role in the prevention of cervical cancer, including recently announced elimination efforts. HPV testing is a recommended approach for cervical cancer screening of women over 30 and for management of those with precancerous cervical lesions. In addition, they are widely used in epidemiological studies, HPV surveillance and vaccination impact monitoring.ObjectivesThe aim was to provide an updated 2020 inventory of commercial molecular HPV tests available on the market.SourcesData were retrieved from internal files, and a detailed search using Medline/Pubmed, Web of Science, Scopus, Google Scholar, Google and Bing, without language or period restrictions, was performed in September 2019 and again in January 2020.ContentWe identified 254 distinct commercial HPV tests and at least 425 test variants available on the global market in 2020, which represents a 31% and 235% increase in the number of distinct tests and variants, respectively, compared with the previous inventory performed in 2015. Although the proportion of commercially available HPV tests with at least one peer-reviewed publication has increased over the past decade, 60% of the HPV tests on the global market are still without a single peer-reviewed publication. Furthermore, 82% of tests lack any published analytical and/or clinical evaluation, and over 90% are not evaluated in line with consensus requirements that ensure safe use in clinical settings.ImplicationsSignificant challenges and scope for improvement still exist for both the HPV scientific community and the manufacturers of HPV tests. The latter must put more effort into validating their products, in agreement with standardized procedures, including all steps of HPV testing and various clinical specimens. High throughput capacity and point-of-care HPV tests are needed, both with affordable prices.     

宫颈癌患者HPV感染和调节性T细胞的相关性研究

目的 探讨宫颈部位人乳头瘤病毒（HPV）感染与CD4＋ CD25＋ CD127-调节性T细胞（TReg）及机体免疫水平的关系.方法 采用第二代核酸杂交扩增技术（HC2）检测41例正常对照组、50例宫颈上皮内瘤变（CIN）Ⅰ、42例CIN Ⅱ～Ⅲ和70例感染HPV宫颈癌患者;采用流式细胞术检测外周血CD4＋CD25＋ CD127-TReg、细胞毒性T细胞（CTL）和NK百分率;采用酶联免疫吸附实验（ELISA）检测血清中TGF-β1和INF-γ的含量.结果 ①HPV感染率随着宫颈病变的加重而升高（P =5.75×10^-19）;②宫颈CIN Ⅱ～Ⅲ组与宫颈癌组外周血CD4＋CD25＋CD127-TReg、CTL、NK、TGF-β1、INF-γ与CIN Ⅰ组及正常对照组比较,差异均 有统计学意义（P=1.03×10^-9）;且在HPV阴阳性组间比较亦有统计学意义（P=2.33×10^-4）;③CTL与NK和Treg呈负相关（P值分别为1.62×10^-6和3.41×10^-5）;INF-γ含量与TReg呈负相关（P=2.11×10^-5）;④HPV与CD4＋ CD25＋ CD127-TReg百分率呈正相关（P =3.02×10^-6）.结论 宫颈HPV感染与TReg密切相关,TReg失调导致的免疫水平下降可能是宫颈癌免疫逃避机制之一.     

Awareness and uptake of human papilloma virus vaccines among female secondary school students in Benin City,Nigeria

BackgroundThere is no Government endorsed HPV vaccine immunisation program in Nigeria. The Vaccine has been available at the University of Benin Teaching Hospital (UBTH) in Benin City for more than 7 years.ObjectivesThe aim was to evaluate awareness about HPV, the prevalence of HPV immunisation and its associated factors among the study population.MethodsA cross-sectional study using interviewer-administered questionnaires among 215 females attending secondary schools in Benin city, Nigeria. Participants were selected using multi-stage stratified sampling. The primary outcome measure was HPV immunisation of the girls.ResultsThe majority of the participants were between 14 to 18 years (58.6%). Almost all the participants (>97%) had not heard of HPV, HPV Vaccines and Cervical cancer. In addition, 2 (0.9%) persons correctly identified that the virus can be transmitted sexually while only 1 person (0.5%) had received the HPV vaccine. The respondents all agreed that they needed to be enlightened about HPV, HPV vaccines and Cervical cancer. Majority (49.3%) of the girls suggested that this could be done through the mass media (49.3%) or their parents (32.1%).ConclusionHPV immunisation, knowledge of HPV vaccines and Cervical cancer among the study population was very low. We recommend interventions in Schools to increase knowledge about cervical cancer and HPV vaccines.     

Functional biomarkers in cervical precancer: An overview

Cervical cancer develops over a long time through precursor lesions that can be detected by cytological screening. Majority of these lesions regress spontaneously. Therefore, the challenge of cervical cancer screening is to detect the lesions that have a high risk of progression. Several promising biomarkers have been described that may improve screening of cervical cancer, but to date, new biomarkers have not been thoroughly validated in high‐quality studies. The knowledge about human papillomavirus as a causative agent of cervical cancer has accumulated over the last decades has opened the possibility to improve the existing prevention strategies and screening practices. p16 has amply been applied on cytologic samples and has been shown to be a promising marker especially in identification of high‐grade dysplasia. ProEx C, a replication marker, has also been recently shown to be a good marker for identification of high‐grade dysplasia and has been used on cytologic samples. Proliferation markers such as MYC, Ki67, telomerase, MCM, topoisomerase 2A and 3q amplification by in situ hybridization technique are other methods being employed in identification of high‐grade dysplasia. However, currently available data on most of the biomarkers does not warrant their routine use yet. This review highlights the major findings of previous studies on cervical cancer biomarkers. Diagn. Cytopathol. 2010;38:618–623. 2009 Wiley‐Liss, Inc.     

Detection and typing of common human papillomaviruses among Jordanian patients

The epidemiology of human papillomaviruses (HPVs) genotype distribution of cutaneous warts in Jordanian patients were studied. A total of 200 samples were collected using skin swabs from patients with warts attending the dermatology clinic at the Jordan University Hospital over the period of June 2010 to October 2010. Another 100 control samples were taken from healthy Jordanian individuals with no current or previous history of warts. DNA extraction and sequencing was carried out using PCR with the FAP primer pair to detect HPV DNA, followed by multiple‐type‐specific (Multiplex) PCR combined with DNA sequencing. The prevalence of HPV among Jordanian patients tested with warts diagnosed clinically was 82% (157/192); of these 45% (87/192) were detected by FAP PCR method, and 37% (70/192) were detected by multiplex PCR method. Sequencing of the FAP positive samples shows that HPV 2 was associated with the highest prevalence (36%), followed by HPV 27 (28%) and HPV 57 (21%). A total of 6% of healthy persons were positive for HPV DNA. In conclusion, this study demonstrates that alpha HPV types (HPV 2, HPV 27, and HPV 57) are associated with the most prevalent cutaneous warts in Jordanian patients J. Med. Virol. 85: 1058–1062, 2013. © 2013 Wiley Periodicals, Inc.     

实时荧光定量PCR法检测人乳头状瘤病毒的实验研究

目的 通过研究病变宫颈中人乳头状瘤病毒(HPV)16/18型的表达,探讨HPV16/18型病毒感染与宫颈病变发生发展之间的关系.方法 结合病理切片诊断,以免疫组化作对照.运用实时荧光定量PCR技术检测病变宫颈中HPV16/18型DNA拷贝数,以及HPV16/18型E7基因mRNA表达量.结果 慢性宫颈炎患者中HPV16/18型感染率低(7.4%).宫颈管上皮内瘤样变(CIN)组HPV16型感染率较高为69.6%,宫颈癌患者巾为72.7%.HPV16型DNA的拷贝数在宫颈上皮内瘤样变患者中与病理分级没有明显的相关性.但在宫颈癌患者中,病毒DNA的拷贝数明显升高,二者差异明显.CIN轻度(I)、中度(Ⅱ)、高度(Ⅲ)组和宫颈癌患者中,HPV16 E7基冈的表达率分别为0、37.5%、42.9%、63.6%.统计学分析表明,HPV16 E7 mRNA的拷贝数与病情呈明显的正相关性.结论 感染者中主要以HPV16型为主,HPV18型较少.宫颈癌患者中HPV16 DNA拷贝数明显高于CIN Ⅱ、Ⅲ组,HPV16型E7 mRNA在宫颈癌中表达率及表达量明显增加并与宫颈癌变呈正相关.实时荧光定量PCR适合临床宫颈病变病毒的筛查与检测.     

Oncogenic activities of human papillomaviruses

Infectious etiologies for certain human cancers have long been suggested by epidemiological studies and studies with experimental animals. Important support for this concept came from the discovery by Harald zur Hausen's group that human cervical carcinoma almost universally contains certain “high-risk” human papillomavirus (HPV) types. Over the years, much has been learned about the carcinogenic activities of high-risk HPVs. These studies have revealed that two viral proteins, E6 and E7, that are consistently expressed in HPV-associated carcinomas, are necessary for induction and maintenance of the transformed phenotype. Hence, HPV-associated tumors are unique amongst human solid tumors in that they are universally caused by exposure to the same, molecularly defined oncogenic agents, and the molecular signal transduction pathways subverted by these viral transforming agents are frequently disrupted in other, non-virus-associated human cancers.     

Detection of Chlamydia trachomatis and herpes simplex virus type 1 or 2 in cervical samples in human papilloma virus (HPV)-positive and HPV-negative women

This study investigated whether the prevalence of human papilloma virus (HPV) in association with Chlamydia trachomatis, herpes simplex virus (HSV)-1 and/or HSV-2 was greater in high-grade than in low-grade or control cervical biopsy specimens. HPV-positive (n = 86) and HPV-negative (n = 213) women were screened for HPV, HSV and C. trachomatis by PCR. The most common HPV genotypes were HPV-16, HPV-6 and HPV-33; mixed HPV infection (n = 12) was also seen. A higher prevalence of C. trachomatis, HSV-1 and HSV-2 was found in HPV-positive samples. High-risk HPV genotypes and combined HPV + C. trachomatis or HPV + HSV-1, but not HSV-2, infections were associated with a greater risk of developing cervical carcinoma.     

134例宫颈上皮内瘤变患者HPV分型检测结果分析

目的探讨宫颈上皮内瘤变(CIN)中不同类型人乳头瘤病毒(HPV)的感染及分型情况。方法回顾性研究北京军区总医院妇科门诊134例已经确诊的CIN患者,采用凯普的核酸分子快速导流杂交基因芯片技术对HPV进行分型。结果 1.各级别CIN以HPV高危亚型单一或者多重感染为主;2.在HPV阳性的101例患者中,单一感染者73例(72.28%),双重感染者24例(23.76%),三重感染者3例(2.97%),四重感染者1例(0.99%);3.HPV-16、58感染率最高,分别为30.83%,24.06%。结论 HPV感染以高危型和多重感染为主;随CIN级别的增高,HPV阳性率也是增高的;HPV-16亚型的感染率最高。     

HIGH‐RISK HPV testing as the primary screening method in an organized regional screening program for cervical cancer: the value of HPV16 and HPV18 genotyping?

Since 2012, testing high‐risk (HR)HPV has been used as the primary screening test for women ≥35 years attending the organized cervical cancer screening program in the city of Tampere. We evaluated the contribution of HPV16/18 genotyping. Data from 2012 and 2013, and the follow‐up samples in 2013 and 2014, respectively, were analyzed. Abbott RealTime High‐Risk HPV test detecting 14 HRHPV genotypes combined with concurrent genotyping for HPV16 and HPV18 was used. HPV was positive in 794 samples out of 11 346 HPV tested women (7%). HPV16/18 was represented in 22% of HPV‐positive cases. Negative cervical cytology (NILM) was reported in 51% of HPV‐positive samples. HPV16/18 genotype was accompanied with 50% of HSIL/ASC‐H cases. The predominance of HPV16/18 in higher grade lesions was even more evident in cervical biopsies as 57% of CIN3 cases were associated with HPV16/18, and only 20% of carcinomas were associated with nonspecified high‐risk (NSHR) genotypes. In agreement with previous studies HPV16/18 genotypes caused higher grade cytological and histological changes/pathologies than NSHR genotypes in primary screening. Nevertheless, the majority of HRHPV genotypes detected in the screened population were nonHPV16/18, and especially within persistent infections, precancerous lesions were found also among women with NSHR genotypes.     

Recombinant vaccines and the development of new vaccine strategies

Vaccines were initially developed on an empirical basis, relying mostly on attenuation or inactivation of pathogens. Advances in immunology, molecular biology, biochemistry, genomics, and proteomics have added new perspectives to the vaccinology field. The use of recombinant proteins allows the targeting of immune responses focused against few protective antigens. There are a variety of expression systems with different advantages, allowing the production of large quantities of proteins depending on the required characteristics. Live recombinant bacteria or viral vectors effectively stimulate the immune system as in natural infections and have intrinsic adjuvant properties. DNA vaccines, which consist of non-replicating plasmids, can induce strong long-term cellular immune responses. Prime-boost strategies combine different antigen delivery systems to broaden the immune response. In general, all of these strategies have shown advantages and disadvantages, and their use will depend on the knowledge of the mechanisms of infection of the target pathogen and of the immune response required for protection. In this review, we discuss some of the major breakthroughs that have been achieved using recombinant vaccine technologies, as well as new approaches and strategies for vaccine development, including potential shortcomings and risks.     

Daily self-sampling for high-risk human papillomavirus (HR-HPV) testing

BackgroundSelf-sampling for HPV as part of primary screening is a well-tolerated method for women not attending organized Pap smear screening and could increase coverage of cervical cancer screening.ObjectiveTo investigate if the prevalence of HR-HPV varies from day to day in infected women and if one single sample is reliable for detecting an ongoing infection.Study designThis is a prospective cohort study on 12 premenopausal and 13 postmenopausal women performing daily self-sampling for HR-HPV testing. They were all HR-HPV-positive 1–3 months ago. Postmenopausal women were sampled for 28 days and premenopausal women sampled during bleeding-free days in one menstrual cycle. A possible difference in viral load between the estrogen-dominated proliferative phase and the progesterone-dominated secretory phase was analyzed.Results and conclusionsConsistent results throughout the sampling period were observed for 19 women, with either a daily presence of HPV (14 women) or no HPV at all during the sampling period (5 women). Of 607 samples from 25 women, 596 were consistently positive or negative for HPV during the sampling period and 11 were inconsistent (2%). There was no difference in HPV copy number between the estrogen dominated proliferative or progesterone dominated secretory menstrual cycle phases.The major finding was a high degree of consistency concerning HR-HPV positivity and negativity of HR-HPV in vaginal fluid during a sustained period of daily self-sampling. It does not appear to matter whether the sample is collected in the proliferative or secretory phase.     

徐州地区女性宫颈感染人乳头瘤病毒基因亚型分析

目的：了解徐州地区不同年龄段女性生殖道人乳头瘤病毒（HPV）感染状况及其分布规律,为徐州地区HPV分子流行病学研究提供依据。方法：以2009年2月～2009年5月来我院就诊的1299例可疑患者为对象,采集其阴道宫颈分泌物标本。用凯普医用核酸分子杂交系统（简称H ybriMax）进行21种HPV（8304、6、11、16、18、31、33、35、39、42、43、45、44、51、52、53、56、58、59、66、68）基因型的分型检测,分析常见感染亚型和分布规律。结果：HPV阳性总检出率为26.02%（338/1299）,高危型主要为HPV16,其余依次为HPV58、52、68、33、53、18感染。低危型主要为HPV11、6、8304感染;〈35岁、35～44岁、〉44岁各年龄组中HPV阳性率分别为26.92%、26.79%、22.27%;宫颈癌患者以HPV16感染为主。结论：HPV6、11、16、58、52、68、33是徐州地区HPV感染的主要型别;不同年龄段HPV阳性率无显著性差异（P〉0.05）;对HPV阳性者进行定期跟踪是防治宫颈癌的重点。     

宫颈瘤变程度与其相对应的HPV基因型分布规律的研究

目的研究不同级别宫颈上皮内瘤变（cervical intraepithelial neoplasia,CIN）与人乳头瘤病毒（Human papil-lomavirus,HPV）基因型分布之间的关系。方法收集321例经组织病理学诊断确定为CIN2及以上（CIN2＋）病人的宫颈分泌物标本,其中CIN2 67例,CIN3/ACIS 247例,浸润性宫颈癌7例。采用深圳亚能生物技术有限公司的人乳头瘤病毒（HPV）基因分型检测试剂盒对所选标本进行HPV检测及基因分型。结果在所有321例标本中,HPV阳性300例,阳性率93.5%,两型及两型以上感染138例,占比43.0%。HPV16最常见,检出率为41.1%（132/321）,其次为HPV31,33,52,18和51。HPV16和33的单型感染在CIN3＋中比CIN2更常见,具有统计学差异（P〈0.05）。在包含有HPV16或HPV33的多型感染中,含有HPV16或HPV33的多型感染在CIN3＋中比CIN2更常见,具有统计学意义（P〈0.05）。结论 HPV16和HPV33比其他型别HPV具有更强的潜在致癌性。     

子宫颈癌治疗性疫苗研究进展

子宫颈癌为妇女最常见的恶性肿瘤之一,其与人乳头瘤病毒(HPV)密切相关.HPV早期基因E6、E7的持续表达是维持癌细胞转化状态所必须的.因此,E6、E7可作为宫颈癌治疗的靶点.目前,针对HPV E66、E7的治疗性疫苗已取得了较大进展.本文就近年来子宫颈癌治疗性疫苗的最新进展进行了综述.     

宫颈癌疫苗的现状和前景

宫颈癌的发病率和病死率较高,宫颈癌疫苗的应用是一种控制宫颈癌发生的重要的预防手段。目前疫苗存在诸多问题,如预防性疫苗对已经感染HPV的患者或确诊为癌前病变及宫颈癌的患者则无治疗作用,且大部分疫苗都只针对该疫苗所包含的HPV特异型产生保护作用等。因此,本文就有关宫颈癌疫苗的临床前和临床研究现状综述如下,并探讨宫颈癌疫苗的临床应用前景。     

HPV感染和p53异常表达与宫颈鳞癌的关系

目的研究宫颈鳞癌与p53蛋白表达和HPV感染的关系,探讨宫颈鳞癌的形成机制。方法采用RT-PCR法分别检测p53基因在39例宫颈鳞癌组织和39例正常宫颈黏膜组织中的表达情况以及PCR方法检测HPV在这些组织中的感染情况。结果 HPV在宫颈鳞癌组中阳性率为48.72%(19/39),正常组织中为14.81%(4/27),p53基因表达在宫颈鳞癌组中阳性率为53.85%(21/39),正常组织中为18.52%(5/27),宫颈癌组的HPV感染和p53表达均高于正常组(P0.05)。结论 HPV感染与p53基因的异常表达与宫颈鳞癌发生密切相关,联合检测能提高准确性。