Evaluation of the Long-Term Anti-Human Papillomavirus 6 (HPV6), 11, 16, and 18 Immune Responses Generated by the Quadrivalent HPV Vaccine

This quadrivalent human papillomavirus (qHPV) (HPV6, -11, -16, and -18) vaccine long-term follow-up (LTFU) study is an ongoing extension of a pivotal clinical study (FUTURE II) taking place in the Nordic region. The LTFU study was designed to evaluate the effectiveness, immunogenicity, and safety of the qHPV vaccine (Gardasil) for at least 10 years following completion of the base study. The current report presents immunogenicity data from testing samples of the year 5 LTFU visit (approximately 9 years after vaccination). FUTURE II vaccination arm subjects, who consented to being followed in the LTFU, donated serum at regular intervals and in 2012. Anti-HPV6, -11, -16, and -18 antibodies were detected by the competitive Luminex immunoassay (cLIA), and in addition, serum samples from 2012 were analyzed by the total IgG Luminex immunoassay (LIA) (n = 1,598). cLIA geometric mean titers (GMTs) remained between 70% and 93% of their month 48 value depending on HPV type. For all HPV types, the lower bound of the 95% confidence interval (CI) for the year 9 GMTs remained above the serostatus cutoff value. The proportion of subjects who remained seropositive based on the IgG LIA was higher than the proportion based on cLIA, especially for anti-HPV18. As expected, the anti-HPV serum IgG and cLIA responses were strongly correlated for all HPV types. Anti-HPV GMTs and the proportion of vaccinated individuals who are seropositive remain high for up to 9 years of follow-up after vaccination.     

Immunogenicity of the Quadrivalent Human Papillomavirus (Type 6/11/16/18) Vaccine in Males 16 to 26 Years Old

Human papillomavirus (HPV) infection can lead to significant disease in males, including anogenital warts, intraepithelial neoplasias, and several types of oral and anogenital cancers. The quadrivalent HPV (type 6/11/16/18) L1 virus-like particle (VLP) vaccine (qHPV vaccine; Gardasil) has recently been demonstrated to prevent persistent infection and associated disease related to vaccine HPV types in males. We report the overall immunogenicity results from a trial of the quadrivalent HPV vaccine in males. Overall, 3,463 heterosexual men and 602 men who had sex with men were enrolled into a randomized, placebo-controlled, double-blind safety, immunogenicity, and efficacy study. Serum samples were collected prior to vaccination at day 1 and at months 7, 24, and 36 postvaccination. Immunogenicity was evaluated with a multiplex, competitive Luminex immunoassay. Almost all subjects (97.4 to 99.2%) seroconverted for vaccine HPV types by month 7. At month 36, 88.9%, 94.0%, 97.9%, and 57.0% of subjects were still seropositive for HPV-6, -11, -16, and -18, respectively. For all vaccine HPV types, black subjects had significantly higher antibody titers at month 7 than did both Caucasian and Asian subjects. An anamnestic antibody response was seen in men seropositive before vaccination. The vaccine was highly immunogenic in males 16 to 23 years of age; responses were comparable to those observed in women. Furthermore, the immune responses were consistent with the established efficacy of the vaccine in the prevention of incident and persistent HPV infection, anogenital warts, and anal intraepithelial neoplasia.     

Comparison of Different Assays To Assess Human Papillomavirus (HPV) Type 16- and 18-Specific Antibodies after HPV Infection and Vaccination

We compared the measurement of human papillomavirus (HPV)-specific serum antibody levels with the virus-like-particle multiplex immunoassay (VLP-MIA), competitive Luminex immunoassay (cLIA), and glutathione S-transferase (GST) L1-based MIA. Using a large panel of serum samples, these assays showed mutually good correlations for both naturally induced and vaccine-derived HPV-specific antibody levels. However, an adaptation of the GST L1-based MIA resulted in an improved correlation with both cLIA and VLP-MIA.     

Vaccination with a Human Papillomavirus (HPV)-16/18 AS04-Adjuvanted Cervical Cancer Vaccine in Korean Girls Aged 10-14 Years

The human papillomavirus (HPV)-16/18 AS04-adjuvanted cervical cancer vaccine has been demonstrated to be highly efficacious and immunogenic with a favorable safety profile. This study assessed the immunogenicity and safety of the HPV-16/18 AS04-adjuvanted vaccine in healthy Korean girls aged 10-14 yr. This multi-center, observer-blind trial randomly assigned 321 healthy girls to receive three doses (0, 1, 6-month schedule) of HPV-16/18 AS04-adjuvanted vaccine or hepatitis A vaccine. Immunogenicity against vaccine antigens was assessed one month post-Dose 3. Solicited and unsolicited adverse events (AEs) and serious AEs (SAEs) were recorded. In the according-to-protocol analysis, all initially seronegative subjects vaccinated with the HPV-16/18 AS04-adjuvanted vaccine had seroconverted at Month 7, with a peak geometric mean titer (GMT) that was 600-fold higher than the natural infection titer of 29.8 EU/mL for HPV-16 and a peak GMT that was 400-fold higher than the natural infection titer of 22.6 EU/mL for HPV-18. The vaccine was well tolerated with no increase in reactogenicity with subsequent doses and no reports of vaccine-related SAEs. In conclusion, the HPV-16/18 AS04-adjuvanted vaccine is shown to be highly immunogenic and generally well-tolerated in Korean girls aged 10-14 yr.     

Prevalence of Anti-Human Papillomavirus Type 16, 18, 31, and 58 Virus-Like Particles in Women in the General Population and in Prostitutes

Genital human papillomavirus (HPV) infection is sexually transmitted. The aim of the study was to characterize serological responses to HPV types 16, 18, 31, and 58 by exploring type-specific virus-like particles (VLPs) in two groups of women with very distinct sexual behaviors. Anti-VLP antibodies for types 16, 18, 31, and 58 and HPV DNA in cervical cells were investigated with 177 prostitutes and 283 age-matched controls from the female general population in Spain. Anti-VLP positivity increased with number of lifetime sexual partners in women from the general population, and no seroresponse was found in virgins. However, in prostitutes HPV infection was characterized by higher multireactivity to three or four VLPs (25%) than the general population (3%) and by a more frequent antibody response to HPV-58 than in the general population. About 75% of the women seropositive for type 58 had been born in a Latin American country. Seroprevalence of HPV and cervical HPV DNA in prostitutes were 14 and 10 times higher than observed in women in the general population (prevalence odds ratio [POR] of HPV seropositivity, 14.04 [95%; CI = 8.4 to 23.6] and POR for HPV DNA, 10.4 [95% CI = 3.9 to 27.6). Our results indicate that prostitutes are at an increased risk of oncogenic HPV infections, and they confirm the validity of anti-VLPs as markers of present or past HPV infection, that the number of sexual partners is the major determinant in acquisition of oncogenic HPV, and that anti-VLPs could be used as a marker of repeated infection in prostitutes.     

Human Papillomavirus (HPV) L1 and L1-L2 Virus-Like Particle-Based Multiplex Assays for Measurement of HPV Virion Antibodies

Humoral immunity to human papillomavirus (HPV) has not been fully characterized, and there is currently no standard serologic test for the measurement of HPV antibodies. Most HPV serologic assays developed to date are based on virus-like particles (VLPs) of the major HPV capsid protein, L1. We sought to compare the performance of a multiplex HPV L1 VLP-based serologic assay to that of an assay based on VLPs comprised of both L1 and the minor capsid, L2. We developed HPV L1 VLP and L1-L2 VLP-based multiplex seroassays for the detection of HPV type 16 (HPV16) and HPV18 virion binding antibodies using Luminex fluorescent bead technology. We compared the performance of these assays to that of established pseudovirion-based neutralization and L1 VLP-based enzyme-linked immunosorbent assays (ELISAs). A total of 391 serum specimens from unvaccinated adult males and females were tested. The L1 and L1-L2 VLP multiplex seroassays each demonstrated substantial agreement with both the neutralization assays and the ELISAs for the detection of HPV16 antibodies (κ = 0.60 to 0.64). However, the L1-L2 VLP seroassay demonstrated better agreement with neutralization assays for the detection of HPV18 antibodies than the L1 VLP seroassay (κ = 0.74 and 0.43, respectively). L1 and L1-L2 VLP seroassays showed excellent agreement with one another for the detection of HPV16 antibodies (κ = 0.86) but only moderate agreement for HPV18 antibodies (κ = 0.44). The HPV L1-L2 VLP seroassay performs well for the concurrent measurement of HPV16 and -18 antibodies in large numbers of samples and may be extended to include other HPV types.     

Simultaneous Quantitation of Antibodies to Neutralizing Epitopes on Virus-Like Particles for Human Papillomavirus Types 6, 11, 16, and 18 by a Multiplexed Luminex Assay

Several different methods have been developed to quantitate neutralizing antibody responses to human papillomaviruses (HPVs), including in vivo neutralization assays, in vitro pseudoneutralization assays, competitive radioimmunoassays (cRIAs), and enzyme-linked immunosorbent assays. However, each of these techniques possesses one or more limitations that preclude testing large numbers of patient sera for use in natural history studies and large vaccine clinical trials. We describe here a new multiplexed assay, by using the Luminex Laboratory MultiAnalyte Profiling (LabMAP3) assay system, that can simultaneously quantitate neutralizing antibodies to human papillomavirus types 6, 11, 16, and 18 in 50 μl of serum. The HPV-Luminex competitive immunoassay measures titers of polyclonal antibodies in serum capable of displacing phycoerythrin-labeled detection monoclonal antibodies binding to conformationally sensitive, neutralizing epitopes on the respective virus-like particles. This competitive Luminex immunoassay was found to be as sensitive, accurate, and precise as the currently used cRIAs. An effective HPV vaccine will most likely require several distinct genotypes to protect against multiple cancer causing papillomaviruses. The HPV-Luminex immunoassay should prove to be a useful tool in simultaneously quantitating antibody immune responses to multiple HPV genotypes for natural history infection studies and for monitoring the efficacy of prospective vaccines.     

Evaluation of a Prototype Real-Time PCR Assay for Carcinogenic Human Papillomavirus (HPV) Detection and Simultaneous HPV Genotype 16 (HPV16) and HPV18 Genotyping

Results from a prototype real-time PCR assay that separately detected human papillomavirus genotype 16 (HPV16), HPV18, and 12 other carcinogenic HPV genotypes in aggregate (cobas 4800 HPV test) and results from a PCR assay that detects 37 HPV genotypes individually (Linear Array) were compared using a convenience sample of cervical specimens (n = 531). The percentage of total agreement between the two assays was 94.7% (95% confidence interval, 92.5 to 96.5%). The Linear Array test was more likely than cobas 4800 HPV test to test positive for the 12 other carcinogenic HPV genotypes among women without evidence of cervical disease (P = 0.004).Persistent cervical infections by approximately 15 carcinogenic human papillomavirus (HPV) genotypes cause cervical cancer and its immediate precursor lesions (17, 22, 27). DNA testing for carcinogenic HPV has proven itself to be more sensitive, albeit slightly less specific, for the detection of cervical intraepithelial neoplasia lesion grade 2 (CIN2) and more severe lesions (CIN2+) (1, 10, 11, 15, 18, 19). Carcinogenic HPV DNA testing of cervical samples has been introduced as an adjunct to cervical cytology into primary cervical cancer screening in the United States (20, 26), and the International Agency for Research on Cancer has endorsed its use as a stand-alone option in primary cervical cancer screening (14).The introduction of carcinogenic HPV DNA testing with cervical cytology as a primary screening modality creates a clinical dilemma—the management of carcinogenic HPV DNA-positive, cytology-negative women (4), who remain at a low but significant risk of CIN2+. One solution for identifying women at risk for CIN2+ in this subgroup is the separate detection of HPV genotype 16 (HPV16) and HPV18, the most carcinogenic HPV genotypes, referring HPV16- or HPV18-positive women immediately to colposcopy and following up with HPV16- and HPV18-negative women after a year (13, 26). While the first generation of clinical HPV tests pooled all carcinogenic HPV genotypes and did not include separate HPV genotyping, at least for HPV16 and HPV18, most of the next generation of clinical tests will offer at least HPV16 and HPV18 genotyping. The utility of full HPV genotyping is less certain (2), as it has been shown that repeatedly testing positive for any carcinogenic HPV is a good proxy for HPV genotype-specific persistence, especially in women aged 30 years and older (7).One of the next-generation tests, the cobas 4800 HPV test, detects a pool of 12 carcinogenic HPV genotypes in aggregate, with concurrent, separate detection of HPV16 and HPV18. We conducted the first evaluation of the cobas 4800 HPV test and compared the results to previously reported results (8) generated by a now well-validated Linear Array HPV genotyping test (5, 6, 9, 12) and to cervical diagnoses.We acquired a convenience sample of 552 anonymized residual PreservCyt (PC; Hologic) specimens, after cytologic interpretation and the histologic diagnosis had been rendered, with human subject research approvals as previously described (3, 8). Women with abnormal cytology, including carcinogenic HPV-positive atypical squamous cells of undetermined significance, underwent colposcopy. Women were assigned a worst histologic diagnosis based on the review of the biopsy specimen and loop electrosurgical excision procedure tissue sample and a pathology review as previously described (3, 8). Five sets of 2-ml aliquots were produced from the residual specimens, balanced for the aliquot order, and were stored at −20°C until used; some sets were missing aliquots because of insufficient specimen volume. Of the 552 PC specimens, nine (1.6%) were excluded because women either had unsatisfactory histology or had disease unrelated to cervical neoplasia (e.g., endometrial carcinoma).In 2006, one set of aliquots was tested for 37 HPV genotypes (genotypes 6, 11, 16, 18, 26, 31, 33, 35, 39, 40, 42, 45, 51 to 56, 58, 59, 61, 62, 64, 66 to 73, 81, 82 subtype [IS39], 82 subtype [W13b], 83, 84, and 89) by using the commercially available Linear Array HPV genotyping test (Roche Molecular Systems) as previously described (5, 8, 12). For Linear Array testing, 285 μl of PC was processed, and the extracted DNA was eluted into 140 μl, of which 50 μl was used in the PCR amplification reaction (0.51% of the 20-ml PC specimen).In 2009, a second set of aliquots was tested for HPV by the cobas 4800 HPV test (Roche Molecular Systems), masked to the results from Linear Array testing. The cobas 4800 system features fully automated sample preparation combined with real-time PCR technology, plus software that integrates the two components. The cobas 4800 HPV test was designed for the amplification and detection of a broad spectrum of high-risk HPV genotypes, along with the coamplification of the human cellular gene β-globin. Briefly, 94 PC aliquots plus a positive and negative control were processed concurrently. HPV and cellular DNA were released by lysing PC aliquots under denaturing conditions at elevated temperatures in the presence of proteinase K, chaotropic agents, and detergents. Isolation and purification of the released nucleic acid occurred on magnetic beads, followed by elution with a low-salt reagent. PCR amplification and detection occurred in a single tube, where probes with four different reporter dyes track the different targets in the multiplex reaction. Reporter dye 1 tracked the high-risk HPV pool with 12 high-risk HPV targets (HPV31, -33, -35, -39, -45, -51, -52, -56, -58, -59, -66, and -68); dyes 2 and 3 tracked HPV16 and -18, respectively; and dye 4 targeted β-globin to provide a control for cell adequacy, extraction, and amplification. Probe cleavage during the amplification process generated an increase in fluorescence, resulting in a sigmoid growth curve in the ideal case. A proprietary algorithm that models the expected curve was employed to generate a cycle threshold (C_T) for determining the presence of HPV or β-globin target. These results were generated with a prototype system where a validated clinical cutoff was not available and thus a conservative cutoff of 45 was used to determine positivity. For the cobas 4800 HPV test, 400 μl of PC was processed, and the extracted DNA was eluted into 150 μl, of which 25 μl was used in the real-time PCR amplification reaction (0.33% of the 20-ml PC specimen).HPV16, -18, -31, -33, -35, -39, -45, -51, -52, -56, -58, -59, -66, and -68 were classified as carcinogenic HPV genotypes. HPV testing results from Linear Array and cobas 4800 HPV tests were also categorized hierarchically according to cancer risk (HPV risk category) as follows: HPV16 > HPV18 > carcinogenic HPV types other than HPV16 and HPV18 > negative for carcinogenic HPV.Seven (1.3%) of the 543 eligible specimens did not have an aliquot for Linear Array testing: three were judged invalid because of the absence of a β-globin signal, and four aliquots were missing. Of the 536 specimens with Linear Array results, two (0.4%) did not have an aliquot for testing by the cobas 4800 HPV test, and three (0.6%) were negative for β-globin by the cobas 4800 HPV test and were excluded from this analysis. This analysis was restricted to women with satisfactory Linear Array and cobas 4800 HPV test results and a final diagnosis which was categorized as normal (which included women with no biopsy specimens taken based on a colposcopic impression of normality) or histologic diagnoses of CIN1, CIN2, or CIN3, or more severe (CIN3+) (n = 531). The hierarchical HPV results for Linear Array and cobas 4800 HPV tests were compared using contingency tables. Kappa value, percentage of total agreement, and percentage of positive agreement were calculated. Differences in HPV categorization and percentage of positive agreement were tested for statistical significance (P < 0.05) using an exact symmetry or McNemar''s χ² test, respectively.The comparison of the results of the Linear Array and cobas 4800 HPV tests, according to the cancer risk categories, is shown in Table ​Table1.1. The percentage of total agreement was 94.7% (95% confidence interval [95% CI], 92.5 to 96.5%), and the kappa value was 0.92 (95% CI, 0.89 to 0.95). There was a significant difference in the categorization of the pooled 12 carcinogenic HPV genotype results (P = 0.003), primarily due to the tendency for the Linear Array test to test positive and the cobas 4800 HPV test to test negative for carcinogenic HPV genotypes other than HPV16 and HPV18. When the results of the tests for all 14 carcinogenic HPV genotypes were pooled, there was 95.5% (95% CI, 93.3 to 97.1%) total agreement and a kappa value of 0.91 (95% CI, 0.87 to 0.94) between the two tests. The Linear Array test was more likely to test positive for any of the 14 carcinogenic HPV genotypes (P = 0.002) than was the cobas 4800 HPV test.

TABLE 1.

Comparison of Linear Array (LA) and cobas 4800 HPV test (c4800) results, categorized hierarchically according to cancer risk^a

Patterns of CD8 T-Cell Epitopes within the Human Papillomavirus Type 16 (HPV 16) E6 Protein among Young Women Whose HPV 16 Infection Has Become Undetectable

The patterns of CD8 T-cell epitopes recognized within the E6 protein in women who had cleared their human papillomavirus 16 infection were examined. T-cell lines were established using autologous dendritic cells infected with a recombinant vaccinia virus. Evidence of potential antigenic epitopes was shown in 8 of 23 (34.8%) women.     

Seroepidemiology of Human Papillomavirus 16 (HPV16) L2 and Generation of L2-Specific Human Chimeric Monoclonal Antibodies

Presently, the seroprevalence of human papillomavirus (HPV) minor capsid antigen L2-reactive antibody is not well understood, and no serologic standard exists for L2-specific neutralizing antibodies. Therefore, we screened a total of 1,078 serum samples for HPV16 L2 reactivity, and these were obtained from four prior clinical studies: a population-based (n = 880) surveillance study with a high-risk HPV DNA prevalence of 10.8%, a cohort study of women (n = 160) with high-grade cervical intraepithelial neoplasia (CIN), and two phase II trials in women with high-grade vulvar intraepithelial neoplasia (VIN) receiving imiquimod therapy combined with either photodynamic therapy (PDT) (n = 19) or vaccination with a fusion protein comprising HPV16 L2, E7, and E6 (TA-CIN) (n = 19). Sera were screened sequentially by HPV16 L2 enzyme-linked immunosorbent assay (ELISA) and then Western blot. Seven of the 1,078 serum samples tested had L2-specific antibodies, but none were detectably neutralizing for HPV16. To develop a standard, we substituted human IgG1 sequences into conserved regions of two rodent monoclonal antibodies (MAbs) specific for neutralizing epitopes at HPV16 L2 residues 17 to 36 and 58 to 64, creating JWW-1 and JWW-2, respectively. These chimeric MAbs retained neutralizing activity and together reacted with 33/34 clinically relevant HPV types tested. In conclusion, our inability to identify an HPV16 L2-specific neutralizing antibody response even in the sera of patients with active genital HPV disease suggests the subdominance of L2 protective epitopes and the value of the chimeric MAbs JWW-1 and JWW-2 as standards for immunoassays to measure L2-specific human antibodies.     

Human papillomavirus (HPV) E6/E7 mRNA as a triage test after detection of HPV 16 and HPV 18 DNA

High‐risk human papillomavirus (HPV) DNA detection provides high sensitivity but low specificity for moderate‐grade cervical intraepithelial neoplasia or worse histological identification. A prospective study evaluated mRNA testing efficacy for predicting this histological diagnosis in case of HPV 16 and/or 18 DNA detection. A total of 165 endocervical samples harboring HPV 16 and/or 18 DNA were tested with NucliSENS‐EasyQ® HPV E6/E7‐mRNA‐assay (Biomerieux, Marcy l´Etoile, France). Women with cytological alterations were referred to colposcopy (n = 111). Moderate‐grade cervical intraepithelial neoplasia or worse was diagnosed in 25.8% of women presenting atypical squamous cells of undetermined significance or low‐grade squamous intraepithelial lesions and in 89.8% of women with high‐grade squamous intraepithelial lesions. mRNA sensitivity was 81.3% and 84.1%, respectively. Specificity was 52.2%, and 80.0%, respectively. Negative predictive value (NPV) was 88.9% in undetermined or low‐grade squamous lesions. Positive predictive value (PPV) was 97.4% in high‐grade squamous lesions. mRNA reduced colposcopies by 44.3% in undetermined or low‐grade squamous lesions. Direct treatment of mRNA‐positive cases reduced 77.5% of colposcopies in high‐grade squamous lesions. Women without cytological alterations were followed for 18 months (n = 35), and moderate‐grade cervical intraepithelial neoplasia or worse was diagnosed in 34.3%; mRNA sensitivity and specificity were 83.3% and 86.9%, respectively. PPV and NPV were 76.9% and 90.9%, respectively for predicting moderate‐grade cervical intraepithelial neoplasia or worse in 18 months. mRNA reduced the number of visits for follow‐up in 62.2%. In conclusion, NucliSENS‐EasyQ® HPV E6/E7‐mRNA‐assay (Biomerieux) can serve as a triage test in case of HPV 16 and/or 18 DNA detection. J. Med. Virol. 85: 1063–1068, 2013. © 2013 Wiley Periodicals, Inc.     

HPV16/18和HPV16E6/E7DNA在宫颈癌组织中的表达

目的检测HPV16/18和HPV16E6/E7 DNA在宫颈癌组织中的表达,探讨其在宫颈癌发病中的作用.方法应用PCR和琼脂糖凝胶电泳方法检测46例宫颈癌组织中HPV16/18和HPV16E6/E7DNA.结果 46例宫颈癌中56.5%(26/46)扩增HPV16/18 DNA,其中宫颈鳞癌25例,宫颈腺癌1例.正常对照组20例HPV16/18DNA均为阴性,与宫颈癌组相比差异有显著性(P<0.01).HPV16/18 DNA阳性拷贝对数值为4.32±2.45.HPV16E6,E7DNA分别有53.8%(14/26)、46.2%(12/26)扩增.结论 HPV16/18和HPV16E6/E7 DNA与宫颈癌的发生密切相关,是宫颈癌恶性转化的关键之一,预示着宫颈癌有较强的增殖能力和转移能力.     

Post Hoc Analysis of the PATRICIA Randomized Trial of the Efficacy of Human Papillomavirus Type 16 (HPV-16)/HPV-18 AS04-Adjuvanted Vaccine against Incident and Persistent Infection with Nonvaccine Oncogenic HPV Types Using an Alternative Multiplex Type-Specific PCR Assay for HPV DNA

The efficacy of the human papillomavirus type 16 (HPV-16)/HPV-18 AS04-adjuvanted vaccine against cervical infections with HPV in the Papilloma Trial against Cancer in Young Adults (PATRICIA) was evaluated using a combination of the broad-spectrum L1-based SPF₁₀ PCR-DNA enzyme immunoassay (DEIA)/line probe assay (LiPA₂₅) system with type-specific PCRs for HPV-16 and -18. Broad-spectrum PCR assays may underestimate the presence of HPV genotypes present at relatively low concentrations in multiple infections, due to competition between genotypes. Therefore, samples were retrospectively reanalyzed using a testing algorithm incorporating the SPF₁₀ PCR-DEIA/LiPA₂₅ plus a novel E6-based multiplex type-specific PCR and reverse hybridization assay (MPTS12 RHA), which permits detection of a panel of nine oncogenic HPV genotypes (types 16, 18, 31, 33, 35, 45, 52, 58, and 59). For the vaccine against HPV types 16 and 18, there was no major impact on estimates of vaccine efficacy (VE) for incident or 6-month or 12-month persistent infections when the MPTS12 RHA was included in the testing algorithm versus estimates with the protocol-specified algorithm. However, the alternative testing algorithm showed greater sensitivity than the protocol-specified algorithm for detection of some nonvaccine oncogenic HPV types. More cases were gained in the control group than in the vaccine group, leading to higher point estimates of VE for 6-month and 12-month persistent infections for the nonvaccine oncogenic types included in the MPTS12 RHA assay (types 31, 33, 35, 45, 52, 58, and 59). This post hoc analysis indicates that the per-protocol testing algorithm used in PATRICIA underestimated the VE against some nonvaccine oncogenic HPV types and that the choice of the HPV DNA testing methodology is important for the evaluation of VE in clinical trials. (This study has been registered at ClinicalTrials.gov under registration no. {"type":"clinical-trial","attrs":{"text":"NCT00122681","term_id":"NCT00122681"}}NCT00122681.)     

Efficacy of Human Papillomavirus 16 and 18 (HPV-16/18) AS04-Adjuvanted Vaccine against Cervical Infection and Precancer in Young Women: Final Event-Driven Analysis of the Randomized,Double-Blind PATRICIA Trial

We report final event-driven analysis data on the immunogenicity and efficacy of the human papillomavirus 16 and 18 ((HPV-16/18) AS04-adjuvanted vaccine in young women aged 15 to 25 years from the PApilloma TRIal against Cancer In young Adults (PATRICIA). The total vaccinated cohort (TVC) included all randomized participants who received at least one vaccine dose (vaccine, n = 9,319; control, n = 9,325) at months 0, 1, and/or 6. The TVC-naive (vaccine, n = 5,822; control, n = 5,819) had no evidence of high-risk HPV infection at baseline, approximating adolescent girls targeted by most HPV vaccination programs. Mean follow-up was approximately 39 months after the first vaccine dose in each cohort. At baseline, 26% of women in the TVC had evidence of past and/or current HPV-16/18 infection. HPV-16 and HPV-18 antibody titers postvaccination tended to be higher among 15- to 17-year-olds than among 18- to 25-year-olds. In the TVC, vaccine efficacy (VE) against cervical intraepithelial neoplasia grade 1 or greater (CIN1+), CIN2+, and CIN3+ associated with HPV-16/18 was 55.5% (96.1% confidence interval [CI], 43.2, 65.3), 52.8% (37.5, 64.7), and 33.6% (−1.1, 56.9). VE against CIN1+, CIN2+, and CIN3+ irrespective of HPV DNA was 21.7% (10.7, 31.4), 30.4% (16.4, 42.1), and 33.4% (9.1, 51.5) and was consistently significant only in 15- to 17-year-old women (27.4% [10.8, 40.9], 41.8% [22.3, 56.7], and 55.8% [19.2, 76.9]). In the TVC-naive, VE against CIN1+, CIN2+, and CIN3+ associated with HPV-16/18 was 96.5% (89.0, 99.4), 98.4% (90.4, 100), and 100% (64.7, 100), and irrespective of HPV DNA it was 50.1% (35.9, 61.4), 70.2% (54.7, 80.9), and 87.0% (54.9, 97.7). VE against 12-month persistent infection with HPV-16/18 was 89.9% (84.0, 94.0), and that against HPV-31/33/45/51 was 49.0% (34.7, 60.3). In conclusion, vaccinating adolescents before sexual debut has a substantial impact on the overall incidence of high-grade cervical abnormalities, and catch-up vaccination up to 18 years of age is most likely effective. (This study has been registered at ClinicalTrials.gov under registration no. NCT001226810.)     

Quadrivalent Meningococcal Vaccination of Adults: Phase III Comparison of an Investigational Conjugate Vaccine,MenACWY-CRM,with the Licensed Vaccine,Menactra

Neisseria meningitidis is a leading cause of bacterial meningitis in the United States, with the highest case fatality rates reported for individuals ≥15 years of age. This study compares the safety and immunogenicity of the Novartis Vaccines investigational quadrivalent meningococcal CRM₁₉₇ conjugate vaccine, MenACWY-CRM, to those of the licensed meningococcal conjugate vaccine, Menactra, when administered to healthy adults. In this phase III multicenter study, 1,359 adults 19 to 55 years of age were randomly assigned to one of four groups (1:1:1:1 ratio) to receive a single dose of one of three lots of MenACWY-CRM or a single dose of Menactra. Serum samples obtained at baseline and 1 month postvaccination were tested for serogroup-specific serum bactericidal activity using human complement (hSBA). The hSBA titers following vaccination with MenACWY-CRM and Menactra were compared in noninferiority and prespecified superiority analyses. Reactogenicity was similar in the MenACWY-CRM and Menactra groups, and neither vaccine was associated with a serious adverse event. When compared with Menactra, MenACWY-CRM met the superiority criteria for the proportions of recipients achieving a seroresponse against serogroups C, W-135, and Y and the proportion of subjects achieving postvaccination titers of ≥1:8 for serogroups C and Y. MenACWY-CRM''s immunogenicity was statistically noninferior (the lower limit of the two-sided 95% confidence interval was more than −10%) to that of Menactra for all four serogroups, with the postvaccination hSBA geometric mean titers being consistently higher for MenACWY-CRM than for Menactra. MenACWY-CRM is well tolerated in adults 19 to 55 years of age, with immune responses to each of the serogroups noninferior and, in some cases, statistically superior to those to Menactra.Neisseria meningitidis is a leading cause of bacterial meningitis in the United States (11). While the incidence of meningococcal disease is highest in infants, the highest case fatality rates are observed among older subjects: 4.6% in children <15 years of age, 22.5% in individuals 15 to 24 years of age, and 16.5% in adults >25 years of age (5). In 2007, approximately 72% of all cases of meningococcal disease in the United States were reported in individuals ≥18 years of age (4a).Two quadrivalent meningococcal vaccines, for prevention of meningococcal disease caused by serogroups A, C, W-135, and Y, are available in the United States: an unconjugated polysaccharide vaccine (MPSV4 [Menomune]; Sanofi Pasteur, Inc., Swiftwater, PA) and a diphtheria toxoid protein conjugated vaccine (Menactra; Sanofi Pasteur, Inc., Swiftwater, PA). In the United States, the quadrivalent meningococcal polysaccharide protein conjugate vaccine is currently recommended for all persons 11 to 18 years of age and for those persons 2 to 55 years of age who are at increased risk of meningococcal disease (4). This vaccine is only available in the United States and Canada. Outside North America, polysaccharide vaccines are the only quadrivalent meningococcal vaccines available.To expand the options for prevention of meningococcal disease, an investigational quadrivalent meningococcal CRM₁₉₇ conjugate vaccine (MenACWY-CRM; Novartis Vaccines, Siena, Italy) was recently developed. Studies have shown that MenACWY-CRM is well tolerated and elicits robust immunogenicity when administered to infants as young as 2 months of age (10, 12), as well as children (2a) and adolescents (9). A large, randomized, controlled, direct comparative phase III study was recently completed, examining the safety and immunogenicity of MenACWY-CRM versus those of Menactra when administered to healthy subjects 11 to 55 years of age. Due to differences in expected immune responses between adolescents and adults, the predefined analyses were powered and analyzed separately for the 11- to 18-year-old and 19- to 55-year-old groups. The larger adolescent age stratum included in the study was powered to demonstrate the consistency of the immune response to three lots of MenACWY-CRM, and the results showed that MenACWY-CRM generated a significantly higher immune response to all four serogroups than Menactra (8). Here, we present the analysis of the safety and immunogenicity of MenACWY-CRM versus those of Menactra among the adult subjects 19 to 55 years of age.     

Testing of a new probe 16/18/45 Hybrid Capture (Digene) in women with high risk HPV infection

BACKGROUND: HPV HR detection test are needed when ASCUS is diagnosed on Pap test. The risk of progression to cervical cancer is dependant on the HPV genotype and three types (HPV 16, 18 and 45) are found in 77.4% of the cervical cancer. Here we have tested a new probe 16/18/45 (Digene) that is able to detect specifically these three types. MATERIAL AND METHODS: Thirty-seven women with a Hybrid Capture 2 High Risk test (Digene) positive were selected to test the new probe 16/18/45. Samples were typed using sequencing reaction after GP5+/GP6+ PCR. Types were given after comparison with the GenBank. Discordant results were controlled with Inno-Lipa HPV genotyping v2 test (Innogenetics). RESULTS: Among the 37 women with HR HPV result, 48.6% were positive with the probe 16/18/45 (18 patients). After genotyping, 12 results were concordant and six discordant (three HPV 31, two HPV 58 et one HPV 59). For the other 19 patients with negative result, 18 are concordant and one discordant (HPV 18). Global concordance for typing between this probe and sequencing was 81% with a kappa test of 0.62 that means a good concordance. Positive predictive value is 66.6% and negative predictive value is 94.7%. CONCLUSION: This study shows a good efficiency of the 16/18/45 probe to detect the genotypes that have the higher risk of progression to cervical cancer. This probe could also allow to follow the epidemiology of HR HPV infection after a large use of HPV vaccines.     

转染HPV16E6基因人树突状细胞疫苗的制备及生物学特性

目的：制备来源于人外周血并转染HPV16E6基因的树突状细胞（DC）疫苗，检测其细胞形态、分子表型及诱导的免疫效应。方法：细胞因子扩增人外周血DC，Lipofectamine转染HPV16E6制备DC疫苗。动态形态学观察，免疫细胞化学及流式细胞术检测分子表达．体外诱导并测定CTL活性。结果：转染DC呈形态迥异的多突起状，其E6蛋白、CD80、CD86和CD83分子的表达率依次为47．3％、82．5％、79．8％和85．7％，诱导杀伤Caski细胞的活性明显高于对照组（P〈0．01）。结论：转染DC疫苗保持了功能成熟DC的形态特征，且内源性表达E6蛋白，能诱导高效的特异性抗肿瘤免疫应答。     

Detection of HPV 16 and HPV 18 DNA in the blood of patients with cervical cancer

Persistent infection of the uterine cervix with high-risk human papillomaviruses (HPV) is causally associated with cancer of the cervix. A few studies have reported the presence of HPV DNA in the blood of women with cervical neoplasia. The aim of this study was to determine if HPV DNA could be detected in whole blood of women with a range of cervical pathologies and with HPV 16 or 18 cervical infections and if there is a correlation between cervical lesion grade and the appearance of HPV DNA in the circulatory system. Forty-five women with histologically graded cervical cancer were confirmed to have cervical HPV 16 or 18 infections. Eleven (24.4%) of these women had detectable HPV 16 or 18 in their blood. The HPV types detected in the blood matched those detected at the cervix. No HPV 16 or 18 DNA was detected in the blood of 32 women with pre-cursor cervical lesions or normal cervical pathology but who had cervical HPV 16 or 18 infections. One of 77 women with normal cervical pathology and no cervical HPV infection was positive for HPV 16 DNA in her blood. The results indicate that HPV DNA can be detected in the blood of women with more advanced cervical carcinomas but not in the blood of women with pre-cursor cervical lesions. The results of our study indicate that the role of HPV DNA in the circulatory system appears not be of diagnostic significance and HPV DNA is only detectable in women with more advanced cervical cancers.