Screening for genital human papillomavirus: results from an international validation study on human papillomavirus sampling techniques.

The objective of this study was to determine the validity of human papillomavirus (HPV) detection using exfoliated cervical cells compared with cervical biopsy specimens in women with normal cervix and to assess whether HPV detection rates using exfoliated cells is dependent on the number and order in which cervical scrapes are taken. Women undergoing hysterectomy for reasons other than cervical cancer were recruited in three hospitals in countries with varying risks of cervical cancer. After informed consent and at the time of surgery, three consecutive cervical scrapes were taken as well as four biopsy specimens, one in each of the quadrants around the cervical os. In this study, 331 women were recruited and provided 992 cell samples and 1324 biopsy samples. All scrapes and a sample of biopsy specimens (n = 103) were tested by polymerase chain reaction enzyme immunoassay using a general primer (GP5+/ bio6+). Type-specific tests were performed for 14 HPV types at the subpicogram level in one test and individually. Positive samples were verified using Southern blot hybridization. The prevalence of HPV DNA was 6.3% in cervical cells. Of 19 HPV positive samples in the scrapes, 17 were confirmed in the biopsy specimens. The agreement, as measured by the Kappa statistic, was 0.90 (P < 0.0001). The concordance in detecting HPV infection between scrapes and biopsy specimens was 97.5%, and the concordance in categorizing the samples as negatives was 94.4%. These values were unchanged when the order in which scrapes were taken was compared. Among women without cervical cancer, HPV DNA detection rates do not vary if exfoliated cells or random biopsy specimens are taken as the primary testing specimen. Screening programs based on highly sensitive HPV DNA detection technology in cell scrapes should expect a minimal underdetection.     

God's gift to women: the human papillomavirus vaccine

An Australian newspaper recently bestowed Ian Frazer the title of "God's gift to women" for his research team's part in developing a vaccine to help control cervical cancer. Here Frazer discusses this work and the science behind the vaccine.     

治疗性人乳头瘤病毒疫苗研究进展

人乳头瘤病毒是以表皮细胞为感染对象的小型DNA病毒,HPV感染与子宫颈癌的发生、发展有着十分密切的关系.宫颈癌位居女性恶性肿瘤的第二位,且呈逐年上升的趋势.已上市的疫苗主要用于预防HPV的感染,对已感染HPV人群的治疗效果欠佳,治疗性疫苗可以通过诱导特异性的细胞免疫应答,阻止甚至清除HPV感染引起的病变及肿瘤.动物实验和临床试验结果显示,治疗性HPV疫苗在治疗HPV感染相关疾病有重要作用.因此,治疗性HPV疫苗的研制及应用已成为近年来研究热点.     

Interlaboratory agreement among results of human papillomavirus type 16 enzyme-linked immunosorbent assays.

Serological assays for measuring antibodies to human papillomavirus type 16 (HPV-16) virus-like particles (VLPs) have become important epidemiologic tools in recent years. However, the interlaboratory replicability of these assays has not been assessed. In this investigation, three laboratories tested a panel of specimens obtained from two different groups: 265 subjects in a vulvar cancer case-control study and 107 healthy volunteer blood donors. Each laboratory used an enzyme-linked immunosorbent assay (ELISA), but no attempt was made to standardize assay procedures among the three laboratories. The data showed good day-to-day intralaboratory replicability in laboratory 1 (correlation coefficient, > or = 0.88) and good intra-assay variability in laboratory 3 (correlation coefficient, > or = 0.93). Interlaboratory correlations, likewise, ranged between 0.61 and 0.80 in both case-control study subjects and healthy blood donors, indicating that ELISA optical density (OD) values between laboratories were linearly related regardless of the population. Kappa coefficients (kappa), based on each laboratory's categorical interpretation of its results (as positive or negative), showed good agreement (kappa, > 0.6) in case-control study subjects and moderate agreement (kappa, > or = 0.4) in blood donors, a population that had few strongly positive sera. When OD values near seropositive cutoffs were treated as indeterminates, there was little discordance between laboratories in either population. The data suggest that each laboratory measured the same humoral immune response and that their HPV-16 VLP ELISAs performed similarly (Pearson correlations). Interlaboratory differences, however, probably due to reagents and procedures, were considerably greater than intralaboratory day-to-day variability. Interlaboratory agreement in determining seropositivity (kappa) could be improved by sharing positive and negative serum controls and by treating marginal results as indeterminate. As part of continuing cooperation to improve interlaboratory agreement, we are preparing bulk serum control specimens to be shared and made available to interested researchers.     

Detection of human papillomavirus type 18 E7 oncoprotein in cervical smears: a feasibility study

Persistent infections by high-risk human papillomaviruses (HPVs) are the main etiological factor for cervical cancer, and expression of HPV E7 oncoproteins was suggested to be a potential marker for tumor progression. The objective of this study was to generate new reagents for the detection of the HPV18 E7 oncoprotein in cervical smears. Rabbit monoclonal antibodies against recombinant E7 protein of HPV type 18 (HPV18) were generated and characterized using Western blotting, epitope mapping, indirect immunofluorescence, and immunohistochemistry. One clone specifically recognizing HPV18 E7 was used for the development of a sandwich enzyme-linked immunosorbent assay (ELISA). The assay was validated using recombinant E7 proteins of various HPV types and lysates from E7-positive cervical carcinoma cells. A total of 14 HPV18 DNA-positive cervical swab specimens and 24 HPV DNA-negative-control specimens were used for the determination of E7 protein levels by the newly established sandwich ELISA. On the basis of the average absorbance values obtained from all 24 negative controls, a cutoff above which a clinical sample can be judged E7 positive was established. Significant E7 signals 6- to 30-fold over background were found in 7 out of 14 abnormal HPV18 DNA-positive cervical smear specimens. This feasibility study demonstrates for the first time that HPV18 E7 oncoprotein can be detected in cervical smears.     

Epidemiology of HIV-1 infection in agricultural plantation residents in Kericho, Kenya: preparation for vaccine feasibility studies

A cross-sectional study was performed to determine the prevalence and risk factors for HIV-1 infection among agricultural plantation residents in Kericho, Kenya. Volunteers were recruited, interviewed, and phlebotomized for HIV-1 serologic testing. Sex-specific adjusted odds ratios were estimated using logistic regression. The overall HIV-1 prevalence was 9.9% (81/820), with prevalence in women more than twice that in men (17.4% vs 8.0%, P=0.001). Among men, elevated HIV-1 prevalence was seen with increasing age, peaking in those older than 30 years (10.3%), marriage (10.4%), Luo tribe affiliation (23.5%), employment (8.9%), travel (11.0%), and being uncircumcised (29.2%). Among women, elevated HIV-1 prevalence was seen in those with no formal education (36.8%) and those who received goods in exchange for sex (36.0%). More than 97% of volunteers expressed a willingness to participate in future HIV-1 studies requiring semiannual visits. HIV prevention efforts have been implemented, along with further research to characterize this population for future cohort feasibility studies and HIV-1 vaccine efficacy trials.     

Reduced acquisition and reactivation of human papillomavirus infections among older women treated with cryotherapy: results from a randomized trial in South Africa

Background  

Treatment of women for high-grade cervical cancer precursors frequently results in clearance of the associated high-risk human papillomavirus (hrHPV) infection but the role of treatment among women without hrHPV is unknown. We investigated whether cervical cryotherapy reduces newly detected hrHPV infections among HIV-positive and HIV-negative women who were hrHPV negative when treated.     

Latent infection induced with cottontail rabbit papillomavirus. A model for human papillomavirus latency.

Latent human papillomavirus infection, a very common event, is most likely the source of primary and recurrent papillomas of the respiratory and genital tracts and might also be the source of neoplastic lesions of the female genital tract and the penis. We have developed a simple model for papillomavirus latency using cottontail rabbit papillomavirus. Skin of domestic rabbits was minimally scarified and inoculated with dilutions of a crude virus suspension ranging from 200 ng to 20 pg viral DNA per inoculated site. Dilution of virus to less than 10 ng/site resulted in delayed and reduced efficiency of inducing warts. After follow-up of 1 to 6 months, sites immediately adjacent to papillomas and inoculated sites where papillomas did not form were biopsied and analyzed by Southern blot and polymerase chain reaction. Inoculated tissues that were clinically and histologically normal contained viral DNA at low levels, detectable by polymerase chain reaction. Ability of the latent virus to induce warts was confirmed by activation with mild skin irritation causing wart formation. This simple model system for latent papillomavirus can be used to study mechanisms of viral activation, therapies to prevent activation, and therapies to eliminate latent virus and thus cure the infection.     

Multimodal e-mental health treatment for depression: a feasibility trial

Background

Internet interventions for depression have shown less than optimal adherence. This study describes the feasibility trial of a multimodal e-mental health intervention designed to enhance adherence and outcomes for depression. The intervention required frequent brief log-ins for self-monitoring and feedback as well as email and brief telephone support guided by a theory-driven manualized protocol.

Objective

The objective of this feasibility trial was to examine if our Internet intervention plus manualized telephone support program would result in increased adherence rates and improvement in depression outcomes.

Methods

This was a single arm feasibility trial of a 7-week intervention.

Results

Of the 21 patients enrolled, 2 (9.5%) dropped out of treatment. Patients logged in 23.2 ± 12.2 times over the 7 weeks. Significant reductions in depression were found on all measures, including the Patient Health Questionnaire depression scale (PHQ-8) (Cohen’s d = 1.96, P < .001), the Hamilton Rating Scale for Depression (d = 1.34, P < .001), and diagnosis of major depressive episode (P < .001).

Conclusions

The attrition rate was far lower than seen either in Internet studies or trials of face-to-face interventions, and depression outcomes were substantial. These findings support the feasibility of providing a multimodal e-mental health treatment to patients with depression. Although it is premature to make any firm conclusions based on these data, they do support the initiation of a randomized controlled trial examining the independent and joint effects of Internet and telephone administered treatments for depression.     

Analysis of human antiviral cytotoxic T-lymphocyte responses for vaccine trials using cryopreserved mononuclear leukocytes: demonstration of feasibility with influenza virus-specific responses.

The feasibility of measuring virus-specific human cytotoxic T-lymphocyte (CTL) activity by using cryopreserved mononuclear leukocytes to support clinical vaccine trials was addressed. Autologous fresh and cryopreserved cells from the same sample of peripheral blood were used as sources of CTL precursors and were tested for influenza virus-specific activity. The data indicated that virus-specific CTL activity could be measured by using cryopreserved cells; this could also be done in assays that are designed to characterize the responsible effector cell population.     

人乳头瘤病毒11型L1/E7嵌合DNA疫苗的构建及其诱导的免疫效应

Objective To construct chimerical DNA vaccine plasmid of human papiUomavirus type 11 (HPV11) L1-E7, and to evaluate its immunogenicity. Methods Molecular cloning techniques were used to construct recombinant plasmid PeDNA3 L1-E7 as a DNA vaccine. BALB/c mice were vaccinated with DNA recombinants through muscle injection. IL-2 and γ-INF secreted by immunized spleens lymphocyte and HPV 11 LI or E7 specific antibodies were assayed by ELISA method. Spleens lymphocyte proliferation was measured by MTT assay. Results The chimerical DNA plasmid of pcDNA3 LI-E7 was constructed correctly. Specific anti-HPV11 E7 and L1 antibodies, specific lymphocyte proliferation and secretions of IL-2 and γ-INF were detected in vaccinated mice. Conclusion Specific immune response, including cellular immunity and humoral immunity, could been detected in mice vaccinated with chimerical DNA vaccine of pcDNA3 L1-E7.     

人乳头瘤病毒11型L1/E7嵌合DNA疫苗的构建及其诱导的免疫效应

Objective To construct chimerical DNA vaccine plasmid of human papiUomavirus type 11 (HPV11) L1-E7, and to evaluate its immunogenicity. Methods Molecular cloning techniques were used to construct recombinant plasmid PeDNA3 L1-E7 as a DNA vaccine. BALB/c mice were vaccinated with DNA recombinants through muscle injection. IL-2 and γ-INF secreted by immunized spleens lymphocyte and HPV 11 LI or E7 specific antibodies were assayed by ELISA method. Spleens lymphocyte proliferation was measured by MTT assay. Results The chimerical DNA plasmid of pcDNA3 LI-E7 was constructed correctly. Specific anti-HPV11 E7 and L1 antibodies, specific lymphocyte proliferation and secretions of IL-2 and γ-INF were detected in vaccinated mice. Conclusion Specific immune response, including cellular immunity and humoral immunity, could been detected in mice vaccinated with chimerical DNA vaccine of pcDNA3 L1-E7.     

人乳头瘤病毒11型L1/E7嵌合DNA疫苗的构建及其诱导的免疫效应

Objective To construct chimerical DNA vaccine plasmid of human papiUomavirus type 11 (HPV11) L1-E7, and to evaluate its immunogenicity. Methods Molecular cloning techniques were used to construct recombinant plasmid PeDNA3 L1-E7 as a DNA vaccine. BALB/c mice were vaccinated with DNA recombinants through muscle injection. IL-2 and γ-INF secreted by immunized spleens lymphocyte and HPV 11 LI or E7 specific antibodies were assayed by ELISA method. Spleens lymphocyte proliferation was measured by MTT assay. Results The chimerical DNA plasmid of pcDNA3 LI-E7 was constructed correctly. Specific anti-HPV11 E7 and L1 antibodies, specific lymphocyte proliferation and secretions of IL-2 and γ-INF were detected in vaccinated mice. Conclusion Specific immune response, including cellular immunity and humoral immunity, could been detected in mice vaccinated with chimerical DNA vaccine of pcDNA3 L1-E7.