Nucleotide Sequence and Phylogenetic Classification of Human Papillomavirus Type 67

The complete nucleotide sequence of human papillomavirus type 67 (HPV 67) cloned from a vaginal intraepithelial neoplasia, has been determined. It consists of 7801 nucleotides with a GC content of 38.4% and exhibits similar genome organizations of genital HPVs. By phylogenetic analysis based on the full nucleotide sequences of E6 open reading frame of 28 genital HPVs, HPV 67 was clustered with HPV 16, 31, 33, 34, 35, 52, and 58. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detection and genotyping of human papillomavirus in cervical samples from Italian patients

Human papillomaviruses (HPVs) are etiological agents of cervical cancer. In order to assess the epidemiological incidence and frequency of different HPV types, we applied a polymerase chain reaction (PCR)-direct sequencing approach based on the use of MY09/MY11 primers as compared to Hybrid Capture assay. Cervical samples were taken from 1,500 women, both with normal and abnormal cytological smears, and we found an incidence of 6.6% of HPV infection in Brescia. Overall, 97 samples tested HPV-positive, yielding 18 HPV types. The four most frequent HPV types were: HPV 16, -31, -6, and -58. This approach could be used in ordinary laboratory settings for quick and reliable typing of known and novel HPVs from clinical specimens and it could also be applied to anti-cancer vaccine development.     

Sequence variation of human papillomavirus type 16 E7 in preinvasive and invasive cervical neoplasias

Variation in the nucleotide sequence of the HPV 16 E7 gene in preinvasive cervical intra-epitherial neoplasia (CIN) and invasive cervical carcinoma specimens was analyzed. Direct DNA sequencing of PCR-amplified products with primers different from those used for PCR with 5-end labeling generated distinct sequence ladders with a low background, even in specimens containing relatively low copy numbers of HPV. Of 14 cervical neoplasias, 11 cases showed sequence diversity from prototype HPV16, and a total of 22 nucleotide exchanges were detected. Nine of these led to single amino acid exchanges: [Thr⁵] to [Lys⁵] in one case and [Asn²⁹] to [Ser²⁹] in eight cases. The [Ser²⁹] E7 was distributed uniformly among invasive carcinomas and precancerous legions, and was also found in a normal cervix. The [Lys⁵] E7 and [Ser²⁹] E7 had transforming potential similar to the prototype E7 assessed by cooperation with the activatedras gene in rat embryo fibroblasts.     

Polymorphism of the capsid L1 gene of human papillomavirus types 31, 33, and 35

The L1 gene encodes for the major capsid protein of human papillomaviruses (HPV). There is limited information on the polymorphism of L1 for types related to HPV‐16. This report explores the polymorphism of L1 in phylogenetically related types 31, 33, and 35 compared to HPV‐16. Genital specimens collected from 732 HIV‐seropositive and 323 HIV‐seronegative women were screened for HPV DNA with consensus L1 PCR. Cervical samples positive for HPV‐16 (n = 74), HPV‐31 (n = 78), HPV‐33 (n = 37), and HPV‐35 (n = 58) were further characterized by PCR‐sequencing of the complete L1 gene. The number of nucleotide substitutions within L1 ranged from 19 for HPV‐33 to 52 for HPV‐31. The ratio of the number of variants/number of isolates tested was higher for HPV‐31 (56.4%, P = 0.05) and HPV‐35 (60.3%, P = 0.04) compared to HPV‐16 (40.5%), while this ratio was lower for HPV‐33 (24.3%), although not significantly (P = 0.14). The maximal distance between HPV variants was greater in the five putative surface‐exposed loops of L1 than in sequences outside the loops (P < 0.01). Synonymous variations were encountered in 1.7% (95% CI 1.1–2.3) of nucleotides inside the L1 loops and 2.4% (95% CI1.2–3.7) of nucleotides outside the L1 loops. Non‐synonymous variations were encountered in 1.8% (95% CI 1.1–2.5) of nucleotides within the L1 loops and 0.2% (95% CI 0–0.4) of nucleotides outside the loops. dN/dS ratios were below 1.0 in extra‐loop and intra‐loop regions, but they were lower in extra‐loop regions. These results suggest that sequences within and outside the hypervariable loops of L1 were under selective constraint. J. Med. Virol. 82: 1168–1178, 2010. © 2010 Wiley‐Liss, Inc.     

Moderate variation of the oncogenic potential among high-risk human papillomavirus types in gynecologic patients with cervical abnormalities

The oncogenic potential of human papillomavirus (HPV) infection was assessed by following the disease course in 455 patients who had had a routine diagnostic Hybrid Capture HPV test due to squamous cell abnormalities of the uterine cervix as detected by cytology and/or colposcopy. At entry, 308 patients had cytologic atypia classified as P3 by the Papanicolau classification, 168 had a positive high-risk HPV test, and 23 were infected only with low-risk HPV. The patients were followed-up using the patient registry until the endpoint of histologically diagnosed cervical intraepithelial neoplasia (CIN). High-grade CIN was diagnosed in 75 surgical biopsies. High-risk HPV infection (relative risk: 76.8 CI(95): 23.7-249.5), cytologic atypia (RR: 16.2 CI(95): 3.9-66.6), and age above 35 (RR: 1.99 CI(95): 1.26-3.16) were independent risk factors for high-grade CIN, while the viral load did not predict oncogenic progression (P = 0.47). After PCR-RFLP typing, the high-risk types were classified into groups as follows: (1) types 16 and 18, (2) types 45, 52, and 56, (3) types 31, 33, 35, 51, and 58. The relative risks of high-grade CIN were 119.1 (CI(95): 36.2-390.9) for group 1, 44.4 (CI(95): 9.8-201) for group 2, and 39.7 (CI(95): 10.9-144.8) for group 3, respectively. The risk ratios between the groups of high-risk types were found to differ at most by a factor of 2.98 (corrected P value: 0.007) indicating that the oncogenic potential varies moderately within the high-risk group of HPVs.     

Orientation-dependent toxic effect of human papillomavirus type 33 long control region DNA in Escherichia coli cells

Virus Genes - The functional analysis of human papillomavirus (HPV) sequence variation requires the molecular cloning of different genomic regions of virus variants. In this study, we report an...     

鲍温氏病组织人乳头瘤病毒16型结构基因L1的克隆及其序列分析

本研究采用ＰＣＲ法从中国妇女鲍温氏病组织标本扩增获得了宫颈癌密切相关的人乳头瘤病毒１６型（ＨＰＶ１６）的晚期基因Ｌ１,并装入测序载体测序分析了其ＤＮＡ的序列,与标准型进行了比较,发现有若干变异,此研究对序列及变异意义进行了分析和讨论,为今后Ｌ１基因分子流行病学调查,Ｌ１蛋白的结构与功能研究、疫苗研制以及ＨＰＶ相关的基础性研究提供了有用的资料,同时在国内外首次报道了鲍温氏病组织中ＨＰＶ１６Ｌ１的序列     

HPV31型L2保守中和表位可诱发广谱中和抗体

目的研究人乳头瘤病毒31型(HPV31)次要外壳蛋白L2保守中和表位的免疫活性及诱发抗体的中和范围。方法合成法获得HPV31 L2 aa.17-40多肽,用EDC法偶联KLH,联合弗氏佐剂免疫新西兰大白兔,用假病毒中和实验检测免疫血清对来自α4、α7、α9、α10及β1亚属的多个HPV型别的中和抗体。结果 HPV31 L2-KLH偶联肽可在新西兰大白兔体内诱发针对至少17种HPV型别的广谱中和抗体,其中HPV31的中和抗体滴度最高,HPV5/45/57的次之。结论首次发现HPV31 L2保守中和表位免疫血清具有广谱中和活性,为基于该表位的广谱HPV疫苗研发奠定了基础。     

Association of human papillomavirus infection with carcinoma of the cervix uteri and its precursor lesions: theoretical and practical implications

Human papillomaviruses (HPVs) are the major aetiological agents of cervical carcinoma. In this review, epidemiological and molecular data are combined to present a model for HPV-induced cervical carcinogenesis. The impact of current knowledge regarding diagnostic and therapeutic approaches is shown, i.e. the use of HPV tests in cervical cancer screening, in the management of atypical smears of uncertain diagnosis and in smears indicative of mild dysplasias, as well as in follow-up examinations during and after therapy. In addition, the value of the two most frequently used HPV detection systems, polymerase-chain reaction (PCR) and hybrid capture (HC) analysis, is discussed. Received: 13 January 2000 / Accepted: 20 March 2000     

Typing of human papillomavirus in women with cervical lesions: Prevalence and distribution of different genotypes

Human papillomavirus (HPV) are distributed widely and persistent infection with high‐risk (HR) HPV is recognized as a necessary cause of cervical cancer. The aim of this study was to evaluate the distribution of different HR‐HPV genotypes in 199 women with cervical pre‐invasive lesions undergoing conservative treatment. A Linear Array HPV Genotyping Test was used to identify individual HPV genotypes in cervical samples. It was observed that the most prevalent HPV genotypes were HPV 16 (52.6%), HPV 51 (13.5%), and HPV 31 (10.9%); HPV 18 was found in 7.3% of the patients. Stratifying the different HPV genotypes according to the severity of the cervical lesion, a strong association between the increasing severity of the histological diagnosis and the detection of more carcinogenic HR‐HPV type was found, and in all but one cervical intraepithelial neoplasia of grade 3 the presence of at least one HR‐HPV could be detected, with more than 70% of cervical intraepithelial neoplasia of grade 3 patients bearing HPV 16. Multiple infections, comprising between 2 and 6 HPV types, were found in 43% of patients; however, the presence of more than 1 HR‐HPV type was not associated with an increased risk of high grade lesions. In conclusion, this data show that HPV 16, 51, 31, 52, and 18 were the prevalent types found in patients with cervical lesion undergoing conservative treatment, with a high prevalence of HPV 16 in cervical intraepithelial neoplasia of grade 3 patients. No association between multiple infection and severity of the lesion could be found. J. Med. Virol. 81:271–277, 2009. © 2008 Wiley‐Liss, Inc.     

Prevalence of human papillomavirus types 6, 11, 16, 18, 31, and 33 in a cohort of Greek women

To study HPV prevalence and HPV types 6, 11, 16, 18, 31, and 33 distribution in cervical smears in a cohort of Greek women. One thousand six hundred thirty-six samples were cytologically evaluated and molecularly analyzed, by PCR based assay. Abnormal cytology was identified in 997 women and 75.4% of them were HPV DNA positive, while 639 had normal cytology and 24.6% were HPV DNA positive. HPV was detected in 62.9% of 256 ASCUS smears, 89.3% of 516 LSIL, 86.7% of 60 HSIL and 47.3% of 165 with cervical carcinoma. Overall, HPV 11 was the most common type (13.4%), followed by 18 (10.3%), 6 (7.2%), 16 (6.4%), 31 (3.4%) and 33 (3.4%). Multiple infections with two (11.3%) or more types, primarily 11 and 18 (4.8%), were also identified. Low-risk types 11 and 6 were common in ASCUS (36.6% and 26.4%, respectively), and high-risk types 16 and 18 in HSIL (42.3% and 30.8%, respectively) and in cancer (51.3% and 41%, respectively). Multiple infections were detected in 2.2% of normal and 31.7% of HSIL. HPV prevalence was 75.4% in abnormal and 24.6% in normal cervical smears. HPV 16 and 18 were the most common types in cancer. Single infection with type 11 and multiple infections with 11 and 18 were more frequent.