Consensus primers targeting human papillomaviruses (HPVs) have biases in sensitivity toward certain HPV types. We applied 3 primer sets (GP51/61, MY09/11, PGMY09/11) in parallel on 120 Chinese cervical cancer specimens. GP51/61 exhibited a poor sensitivity for HPV52, for which the prevalence among squamous cell cervical cancer was underestimated from 14.6% to 0%. The fact that HPV52 should rank second in prevalence among squamous cell cervical carcinoma in Hong Kong could be missed if GP51/61, a worldwide commonly used primer set, was selected for HPV detection. Biases in HPV type-specific sensitivity may result in misprioritization of vaccine candidates. ' 2005 Wiley-Liss, Inc.Key words: human papillomavirus; detection; genotype; vaccine; Chinese; Hong Kong; PCR Cervical cancer is the second most common cancer in women worldwide. Strong and consistent evidence accumulated over the past 2 decades confirms the aetiologic role of human papillomaviruses (HPVs) and provides a strong impetus for developing HPV vaccines to prevent cervical cancer. More than 100 HPV types have been identified, and at least 30 have been found in cervical cancers. 1 Given the diversity of HPV types and the largely typespecific immunity after natural infections, 2-4 it is important to delineate the prevalence of different HPV types found in cervical cancers so as to guide the selection of vaccine candidates. Most studies on HPV have employed consensus primers with an intention to cover a broad spectrum of HPV types. With the more than 10% sequence variation between HPV types, biases in sensitivity of a given primer set toward certain types could happen. In our study, we examined the influence of detection methods on assessing the prevalence of HPVs and thus their priority as cervical cancer vaccine candidates. Material and methodsA total of 120 cervical cancer specimens (105 fresh frozen and 15 paraffin embedded; 89 squamous cell carcinoma, 26 adenocarcinoma, 4 adenosquamous carcinoma and 1 lymphoepitheliod carcinoma) collected from Hong Kong Chinese aged 26-84 years (mean 55 years; SD 13.9) were examined. Total DNA was extracted by the QIAamp DNA mini kit (QIAGEN, Hilden, Germany) and with the quality of extracted preparations confirmed by beta-globin PCR. 5 The clinical materials were collected with a written informed consent. Our study was approved by the local institutional ethics committee, and the human experimentation guidelines of the local institute were followed in the conduct of our study.HPV detection and typing was accomplished by 3 different methods in parallel. In the first method, HPV DNA was amplified by the GP51/61 primers that target an approx. 150 bp fragment of the L1 region. 5,6 The HPV type was identified by direct sequencing of PCR amplicons. In the second method, the MY09/ 11 primers that target an approx. 450 bp fragment of the L1 region was used for PCR. 7,8 HPV type was identified by restriction fragment length polymorphisms (RFLPs) using endonucleases RsaI and DdeI as previously described. 8 Ambiguous RFLPs w...
A population-based study was conducted on 256 southern Chinese with cervical intraepithelial neoplasia grade III (CIN III) or invasive cervical cancer (ICC) and on 258 controls to examine the associations between HLA-B alleles, infection with high-risk human papillomaviruses (HPVs) and the development of cervical neoplasia. HLA-B15 was found to be protective for CIN III/ICC overall (p corrected 5 0.003), and for HPV52-positive CIN III/ICC (p corrected 5 0.003). A marginal protective effect of B15 was observed for HPV16-positive CIN III/ICC, but no significant associations were revealed for HPV18-or HPV58-positive cases. None of the HLA-B alleles were found to confer an increased risk for cervical neoplasia. HLA-B15 is common among Asian for whom HPV52, a worldwide uncommon HPV type, also exists in a relatively high prevalence. It would also be worthwhile to assess the association between HLA-B15, HPV52 and cervical cancer in other Asian populations. ' 2005 Wiley-Liss, Inc.Key words: HLA; Chinese; HPV; cervical cancer; host genetics Infection with high-risk human papillomaviruses (HPVs) is necessary for the development of cervical cancer.1 However, HPV infection alone is not sufficient and most HPV infections regress spontaneously. For cases that do progress to cancer, a long period of latency is usually required. It has become evident that a combination of environmental, viral and host factors interplay to influence the malignant progression.2 The human leukocyte antigen (HLA) system is involved in presenting foreign antigens to immune cells. This family of molecules is divided into 2 major classes. The class I molecules including A, B and C are expressed in most nucleated cells and are essential for targeting infected cells for killing by cytotoxic T-cells. HLA polymorphism results in variation of the peptide binding cleft that influences the immune response to pathogens. Certain HLA alleles have shown to be associated with an increased or decreased risk for cervical cancer among different ethnic populations. 4 Although cervical cancer remains an important cause of death in most parts of China, so far little is known on its association with HLA alleles. The objective of our study was to assess the association between HLA alleles and high-risk HPV infections on the risk of development of cervical neoplasia in southern Chinese. Our study focused on the HLA class I B alleles, as most previous studies have been on class II alleles. Material and methods Case subjectsA population-based study was performed to compare the frequency of HLA-B alleles between Hong Kong Chinese women with cervical neoplasia and controls. Women attending the Gynaecology Clinic at the Prince of Wales Hospital with biopsy-confirmed cervical intraepithelial neoplasia (CIN) III or invasive cervical carcinoma (ICC) were recruited. Peripheral blood samples and cervical scrape/biopsy samples were collected from participants with a written informed consent. Women with surnames that appeared to be non-Chinese were excluded from the study. The study...
Multiple determinants are involved in the progression of human papillomavirus (HPV)-infected cervical lesion to invasive cancer. Human leukocyte antigen (HLA) polymorphism seems to play a role. This study examined the association between HLA-DRB1 polymorphism, high-risk HPV infection, and the development of cervical neoplasia in southern Chinese. Three hundred and seventy women with cervical neoplasia (43 cervical intraepithelial neoplasia grade II, 154 grade III, and 173 invasive cancers) and 323 controls were recruited for HLA-DRB1 typing by a sequence-based approach. Cervical specimens were collected for HPV detection by a consensus primer-based polymerase chain reaction, and with the type of HPV identified by hybridization with type-specific oligonucleotide probes. A protective effect of HLA-DRB1*12 for cervical neoplasia was observed, and with stronger associations when subgroup analyses were carried out for patients infected with HPV16 and HPV58. The protective effect of HLA-DRB1*13 that had been reported from other populations was not observed. The data obtained in this study showed that HLA-DRB1*03 conferred a higher risk for HPV18-infected, but not for HPV16-, HPV52-, or HPV58-infected cervical lesions. Although, HPV52 was reported as uncommon worldwide, it was found to be the second most prevalent type in the southern Chinese population. However, no additional risk association was observed when subgroup analyses were performed for HPV52-infected patients. The current study shows that, among southern Chinese, the outcome of HPV-infected cervical lesions is associated with HLA-DRB1 polymorphism. These associations often vary with the type of HPV infection.
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