Low-risk Human Papillomavirus Types Research Articles

Human Papillomavirus Genotype Distribution in a Hospital-based Single-Center Study

Background: Cervical cancer is reported as one of the most prevalent malignancies worldwide in women. There is a substantial connection between cervical diseases and infection with Human Papillomavirus (HPV), especially Types 16 and 18 of HPV. Objective: This study aimed to assess the HPV genotype distribution to determine its most common type at a referral hospital in Iran. Methods: This cross-sectional study was conducted on 400 women with positive HPV result tests who visited the gynecologic oncology clinic of Baqiyatallah Hospital from 2017-2021. Results: The average age of subjects was 37.46±9.75 years (range=19-74). About 167 (41.8%) of the patients were between 25 to 35 years. The analysis showed that 137 (34.2%) patients had only low-risk (LR) typing, 129 (32.2%) patients had only high-risk (HR) typing, and 134 (33.5%) patients had both HR and LR typing. The most frequent LR HPV types were HPV 6 (18.5%) and 11 (17%), and the most frequent HR HPV types were HPV-16 (14.7%), 52 (8.2%), 18, and 31 (6.5%). Conclusion: This study showed that the most frequent age category in both low- and HR HPVpositive women is the age 25 to 35 years. The second most common HR-HPV subtypes included HPV-52, not HPV-18, which indicates variations in HR-HPV subtypes in different populations and races and warrants further research.

Human Papillomavirus Carcinogenicity and the Need of New Perspectives: Thoughts from a Retrospective Analysis on Human Papillomavirus Outcomes Conducted at the Hospital University of Bari, Apulia, Italy, between 2011 and 2022.

The current manuscript's aim was to determine the human papillomavirus (HPV) genotype-specific prevalence and distribution among individuals, males, and females, of different ages in the region of Apulia, Italy, highlighting the possible variables involved in the carcinogenicity mechanism. In addition, we proposed two hypothetical models of HPV's molecular dynamics, intending to clarify the impact of prevention and therapeutic strategies, explicitly modeled by recent survey data. We presented clinical data from 9647 participants tested for either high-risk (HR) or low-risk (LR) HPV at the affiliated Bari Policlinic University Hospital of Bari from 2011 to 2022. HPV DNA detection was performed using nested-polymerase chain reaction (PCR) and multiplex real-time PCR assay. Statistical analysis showed significant associations for all genders and ages and both HR- and LR-HPV types. A major number of significant pairwise associations were detected for the higher-risk types and females and lower-risk types and males. The overall prevalence of HPV was 50.5% (n-4.869) vs. 49.5% (n-4.778) of the study population, of which 74.4% (n-3621) were found to be HPV high-risk (HR-HPV) genotypes and 57.7% (n-2.807) low-risk HPV (LR-HPV) genotypes, of which males were 58% and females 49%; the three most prevalent HR-HPV genotypes were HPV 53 (n707-15%), 16 (n704-14%), and 31 (n589-12%), and for LR-HPV, they were 42 (19%), 6 (16%), and 54 (13%); 56% of patients screened for HPV were ≤ 30 years old, 53% were between 31 and 40 years old, 46% were 41-50 and 51-60 years old, and finally, 44% of subjects were >60 years old. Our study provided comprehensive epidemiological data on HPV prevalence and genotype distribution among 9647 participants, which could serve as a significant reference for clinical practice, and it implied the necessity for more effective screening methods for HPV carcinogenesis covering the use of more specific molecular investigations. Although this is a predominantly descriptive and epidemiological study, the data obtained offer not only a fairly unique trend compared to other studies of different realities and latitudes but also lead us to focus on the HPV infection within two groups of young people and adults and hypothesize the possible involvement of dysbiosis, stem cells, and the retrotransposition mechanism.

Seegene Anyplex II assays detect HPV consistently using DNA extracts from different extraction methods.

The efficiency of PCR-based diagnostic assays can be impacted by the quality of DNA template, and anal samples can be particularly problematic due to the presence of faecal contaminants. Here, we compared the Quick-DNA Viral Kit (Zymo, Zymo Research, CA) and MagNA Pure 96 DNA and Viral NA Small Volume Kit (MP96, Roche) for use of the Seegene Anyplex II HPV28 assay (Anyplex28, Seegene) with anal samples. A total of 94 anal samples extracted using the MP96 and Zymo kits were tested via the Anyplex28, which detects high-risk human papillomavirus (HR-HPV, Panel A) and low-risk (LR-HPV, Panel B) HPV types. Testing the HR-HPV types (Panel A), 86 (91.5%) MP96 and 84 (89.4%) Zymo samples were deemed assessable. Overall agreement between the two methods was 87/94 (92.6%, 95% CI: 85.3-97.0) with the Kappa value of 0.678 (0.5-0.9). Of the 87 assessable samples, 50 (57.5%) were concordant, 34 (39.1%) partially concordant, and 10 (11.5%)discordant. In conclusion, the Anyplex28 produces comparable HPV genotyping results when using DNA extracts from either of these two methods.

Human papillomavirus infection: Epidemiology, biology, host interactions, cancer development, prevention, and therapeutics.

Human papillomavirus (HPV) infection is one of the most common sexually transmitted infections worldwide. It is caused by the HPV, a DNA virus that infects epithelial cells in various mucous membranes and skin surfaces. HPV can be categorised into high-risk and low-risk types based on their association with the development of certain cancers. High-risk HPV types, such as HPV-16 and HPV-18, are known to be oncogenic and are strongly associated with the development of cervical, anal, vaginal, vulvar, penile, and oropharyngeal cancers. These types of HPV can persist in the body for an extended period and, in some cases, lead to the formation of precancerous lesions that may progress to cancer if left untreated. Low-risk HPV types, such as HPV-6 and HPV-11, are not typically associated with cancer but can cause benign conditions like genital warts. Genital warts are characterised by the growth of small, cauliflower-like bumps on the genital and anal areas. Although not life-threatening, they can cause discomfort and psychological distress. HPV is primarily transmitted through sexual contact, including vaginal, anal, and oral sex. It can also be transmitted through non-penetrative sexual activities that involve skin-to-skin contact. In addition to sexual transmission, vertical transmission from mother to child during childbirth is possible but relatively rare. Prevention of HPV infection includes vaccination and safe sexual practices. HPV vaccines, such as Gardasil and Cervarix, are highly effective in preventing infection with the most common high-risk HPV types. These vaccines are typically administered to adolescents and young adults before they become sexually active. Safe sexual practices, such as consistent and correct condom use and limiting the number of sexual partners, can also reduce the risk of HPV transmission. Diagnosis of HPV infection can be challenging because the infection is often asymptomatic, especially in men. In women, HPV testing can be done through cervical screening programs, which involve the collection of cervical cells for analysis. Abnormal results may lead to further diagnostic procedures, such as colposcopy or biopsy, to detect precancerous or cancerous changes. Overall, HPV infection is a prevalent sexually transmitted infection with significant implications for public health. Vaccination, regular screening, and early treatment of precancerous lesions are key strategies to reduce the burden of HPV-related diseases and their associated complications. Education and awareness about HPV and its prevention are crucial in promoting optimal sexual health. This study aimed to carry out a literature review considering several aspects involving HPV infection: Global distribution, prevalence, biology, host interactions, cancer development, prevention, therapeutics, coinfection with other viruses, coinfection with bacteria, association with head and neck squamous cell carcinomas, and association with anal cancer.

Prevalence of Trichomonas Infection in Relation to Human Papillomavirus (HPV) in Pap Smear Samples of Female Patients Referred to Shahid Sadoughi Hospital, Yazd (Iran).

Objectives Human papillomavirus (HPV) and Trichomonas vaginalis (TV) infections have been proposed as risk factors for cervical cancer. This study has been conducted with the aim of investigating the prevalence of TV and its relationship with HPV in women who underwent Pap smear testing as part of cancer screenings. Materials and methods The sampling of liquid-based cervical tissue was conducted among 500 women referred to the women's clinic of Shahid Sadoughi Hospital in Yazd, Iran. The studied samples were examined for Pap smear tests and microscopic identification of TV, as well as HPV-DNA detection and the determination of high-risk and low-risk HPV types by the polymerase chain reaction (PCR) method. The results were analyzed using the IBM SPSS Statistics for Windows, Version 24 (Released 2016; IBM Corp., Armonk, New York) software. Results The individuals included in the study were 16-72 years old. The prevalence rate of TV infection in this population was found to be 29.2%, and the frequency rate of HPV was reported to be approximately 19.4%, with high-risk HPV, including HPV-56, having the highest frequency. The Pap smear test results were reported as abnormal in 20.2%, anda significant correlation was observed between HPV infection and an abnormal Pap smear test (P < 0.05). In addition, a notable correlation was detected between TV infection and high-risk and low-risk HPV (P < 0.05). Conclusion According to the significant relationship found between the two pathogens, TV and HPV, in the abnormal Pap smear test results, TV infection can be considered a risk factor for HPV infection, as well as uterine lesions and cancer.

Abstract 4804: Prevalence of anal HPV infection in women attending a gynecology and colposcopy clinic: Assessing the potential impact of HPV vaccination

Abstract Human papillomavirus (HPV) accounts for over 80% of anal cancer cases. In the effort to prevent HPV-related cancers, the 9-valent HPV vaccine emerged as a crucial intervention, targeting HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58; the previous 4-valent (types 6,11,16,18) and 2-valent (types 16, 18). This study involved 213 women aged 21-50 attending gynecology and colonoscopy clinics in San Juan, Puerto Rico. The study assessed anal HPV prevalence and analyzed vaccination status-related differences. Anal swabs were collected for HPV testing using the ATILA AmpFire, targeting 17 HPV types. This method identified 15 high-risk HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, and 68) and two low-risk HPV types (6 and 11). Prevalence of any HPV, HR-HPV, and LR-HPV, and of specific HPV types were calculated by overall and vaccination status. Differences were assessed for statistical significance (p&amp;lt;0.05) using Fisher's exact or Pearson χ2 test. The mean age of women was 36.0 ± 5.9 years. Overall, 19.7% (n=44) had received the HPV vaccine; among them, 52.2% (n=23) had received the 4-valent vaccine, 2.3% had received the 2-valent vaccine, 2.3% had received the 9-valent vaccine, and 43.2% were uncertain about their specific HPV vaccine type. The prevalence of anal HPV infection was 68.5% for any HPV, 61.5% for HR-HPV, and 15.0% for LR-HPV. The most prevalent HR-HPV types were HPV 56 (13.2%), HPV 68 (11.7%), and HPV 66 (11.2%). There were no statistically significant differences in the prevalence of any HPV, HR-HPV, and LR-HPV between vaccinated and unvaccinated women. The most common HR-HPV type among vaccinated were HPV 66 (20.5%), HPV 31 (18.2%), HPV 56 (15.9%), while the most common among unvaccinated women were HPV 56 (12.4%), HPV 68 (12.4%), and HPV 16 (12.4%). Additionally, when evaluating as a grouped variable, the HPV genotypes covered by the 4-valent, unvaccinated women had a significantly higher prevalence of these types as compared to those vaccinated (28.9% vs. 13.6%, respectively, p-value =.03). The unvaccinated women showed a lower prevalence for HPV types covered by the 9-valent vaccine compared to the vaccinated cohort (43.2% vs. 45.5%). However, this difference was not statistically significant. We found a high prevalence of anal HPV among these women, with lower prevalence of 4-valent and 2-valent HPV types among vaccinated women, evidencing the pivotal role of HPV vaccination. Future research should assess the 9-valent vaccine's long-term effectiveness against more HPV types. Funding: Research Centers in Minority Institutions (RCMI), University of Puerto Rico (NIMHD # 2U54MD007600-36), and CAPAC Research Training Program (NCI #R25CA240120). Citation Format: Anelisse Dominicci-Maura, Ana Patricia Ortiz, Josefina Romaguera, Filipa Godoy-Vitorino. Prevalence of anal HPV infection in women attending a gynecology and colposcopy clinic: Assessing the potential impact of HPV vaccination [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4804.

Comparison of nylon-flocked swabs and cotton swabs in the detection of human papillomavirus infection in men.

Human papillomavirus (HPV) is an oncogenic virus and the commonest sexually transmitted pathogen worldwide. Appropriate sampling is an important factor in infection management. This study aimed to compare the efficacy of cotton swabs (CS) and nylon-flocked swabs (NFS) in sampling for HPV-DNA PCR testing in male patients with genital warts. The study included men with genital warts who presented to the urology outpatient clinic of Antalya Medical Park Hospital. Before wart treatment, multisite sampling of the penis and genital area was performed separately with CS and NFS. The samples were analyzed for HPV-DNA using real-time PCR. The study included 45 men with a mean age of 32.1 ± 8.6years. At least one HPV type was detected in all 45 patients with NFS sampling and 44 patients with CS sampling (total HPV types detected: 106 and 84, respectively). NFS sampling detected 52 high-risk HPV types in 37 of the 45 patients, while CS sampling detected 37 high-risk types in 19 patients (p = 0.029). NFS sampling also detected a total of 54 low-risk HPV types in all 45 patients, versus 47 low-risk HPV types in 41 patients with CS sampling. Multiple HPV types were detected in 30 patients with NFS and 17 patients with CS (p = 0.001). NFS were more effective than CS for HPV-DNA testing in men with genital warts. NFS were superior to CS in detecting multiple-type HPV infection and high-risk HPV types. The use of NFS should be recommended for HPV-DNA PCR testing in men.

Feasibility and Acceptability of Self-sampling for Human Papillomavirus DNA Testing Among Asymptomatic Women in Rural Delta State, Nigeria

Background: Self-sampling and human papillomavirus (HPV) DNA testing can be vital tools in cervical cancer screening for hard-to-reach women with limited access to health care in sub-Saharan Africa. This study explored the feasibility and acceptability of self-sampling for HPV testing among asymptomatic women in Orhuwhorun community in Delta State. Methods: This was a cross-sectional study of 230 women aged between 30 - 65years, enrolled by a multi-stage sampling method. Recruited women were asked to provide self-collected vaginal samples between May and June, 2021. HPV detection and genotyping was done using 21 HPV Geno-Array Diagnostic kit. Participants were asked to complete questionnaires after self-sampling to point out their experiences and acceptability of HPV self-sampling. Results: An excellent feasibility of self-sampling (95.2%) was observed. The acceptability rate of self-sampling was 93.0%, and 99.6% (229/230) of the participants were confident that they used the device correctly. The quality of self-sampling was satisfactory in 100% of the samples; 21.1% (48/228) of the samples were positive for HPV, including 12.3% (28/228) with high-risk HPV types,2.6% (6/228) with probable high risk HPV types and 1.8% (4/228)with low-risk HPV types. Multiple HPV infection occurred in 10 cases (4.4%). Conclusion: This study indicates that self-sampling is a feasible and acceptable approach for cervical cancer screening among women in rural Delta State. Thus, the government should consider self-sampling as a valuable strategy in implementing national cancer screening programme.

Vaccine and Non-Vaccine HPV Types Presence in Adolescents with Vertically Acquired HIV Five Years Post Gardasil Quadrivalent Vaccination: The ZIMGARD Cohort.

Human papillomavirus (HPV) vaccination programs are a key intervention in protecting individuals against HPV-related disease. HIV1-infected individuals are at increased risk of HPV-associated cancers. This study was conducted to evaluate the potential role of prophylactic HPV vaccines in preventing new HPV infections among participants with perinatally acquired HIV who received the quadrivalent HPV vaccine at least five years before this study. This cross-sectional study was conducted at Newlands Clinic, Harare, Zimbabwe. The clinic provided the Gardasil quadrivalent HPV vaccine (4vHPV) to 624 adolescents living with HIV starting in December 2015. Vaginal and penile swabs were collected and tested for HPV types from the study participants who had received the 4vHPV vaccine 5-6 years before enrolment. We present the results of 98 participants (44.6% female) vaccinated at a median age of 15 years (IQR 12-16). The mean amount of time since vaccination was 6 years (SD: ±0.4). The HPV-positive rate amongst the analyzed swabs was 69% (68/98). Among 30/98 (31%) HPV-positive participants, 13/98 (13%) had low-risk HPV types, and 17/98 (17%) had high-risk HPV types. Twelve participants tested positive for HPV18, only one participant tested positive for HPV16, and an additional four (4.3%) tested positive for either type 6 or 11, with respect to vaccine-preventable low-risk HPV types. The Gardasil quadrivalent HPV vaccine (4vHPV) was expected to protect against infection with HPV types 16, 18, 6, and 11. We demonstrated a possible waning of immunity to HPV18 in 17% of the participants, and an associated loss in cross-protection against HPV45. We observed a relatively high prevalence of 'opportunistic non-vaccine HPV types' or 'ecological niche occupiers' in this cohort, and suggest further research on the involvement of these types in cervical and other genital cancers. Our study is one of the few, if not the first, to report on HPV vaccine immunoprotection among people living with HIV (PLWH), thereby setting a baseline for further studies on HPV vaccine effectiveness among PLWH.

Detection and typization of HPV genotypes in subjects with oral and upper respiratory tract lesions, Milan, Italy

Background Oral human papillomavirus (HPV) is common among healthy individuals but causes and implications of persistent infections are under evaluation in the pathogenesis of head and neck neoplasms. Methods This was a retrospective study evaluating the prevalence of high-risk (HR), probable HR and low-risk (LR) HPV types in patients reporting signs/symptoms of oral and upper respiratory tract lesions. Individuals attending between 2019 and 2022 a University Hospital in Milan, Italy, with risk factors for HPV (unprotected oral sex and/or previous documentation of HPV infection in oral and upper respiratory tract and/or another anatomical site) were included. Results Fourteen out of 110 (12.7%) individuals tested positive for HPV DNA. The prevalence of HR-HPV and LR-HPV was 3.6% (4/110) and 9.1% (10/110), respectively. No probable/possible HR-HPV was detected. Specifically, 10/110 (9.1%) were diagnosed with 1 LR-HPV genotype, 3/110 (2.7%) were infected with 1 HR-HPV and 1/110 had 3 concomitant HR-HPV types. HPV 16 (2.7%, 3/110) and 6 (4.5%, 5/110) were the most common HR and LR types, respectively. One patient positive for HPV 16, 33 and 35 was diagnosed with cancer at the base of the tongue. Two individuals among those who tested positive for HPV DNA reported previous HPV vaccination. Conclusions Our data, in line with observations from previous prevalence studies, support the potential role of HPV in head and neck neoplasms. HPV DNA testing should be performed in patients presenting lesions in oral/respiratory tracts and risk factors for HPV. Improvement in HPV vaccination coverage is warranted.

A Low-Risk HPV-Associated Well-Differentiated Squamous Cell Carcinoma of the Cervix with Low-Grade Squamous Intraepithelial Lesion Morphology: Clinical and Pathologic Diagnostic Difficulties and Review of the Literature.

Approximately 95% of cervical squamous cell carcinomas are associated with high-risk HPV, with a small number of HPV-independent tumors. However, low-risk HPV types have also been detected in rare cervical squamous cell carcinomas. Low-grade squamous intraepithelial lesion-related changes are a rare morphologic finding in cervical squamous cell carcinoma. We present the case of a 30-yr-old woman who presented with pelvic pain and foul-smelling vaginal discharge showing an exophytic lesion protruding from the cervix. Repeated superficial biopsies showed a low-grade squamous intraepithelial lesion (LSIL) characterized by binucleation and koilocytosis. Chromogenic in-situ hybridization revealed the presence of HPV6/11. The absence of high-risk HPV was confirmed by PCR. After following the patient for nine months without intervention, type III hysterectomy and bilateral pelvic paraaortic lymphadenectomy were performed. Microscopic examination showed well-differentiated squamous cell carcinoma with solid epithelial islands and extensive eosinophilic cytoplasm without pleomorphism. HPV 6 and 11 were also detected with chromogenic in-situ hybridization. Neoplasm invaded the full-thickness of the cervical wall and infiltrated the vagina, parametrium, the proximal ureter and bladder. The patient who received chemoradiotherapy is disease-free at 36 months follow-up. Low-risk HPV-related well-differentiated invasive squamous lesions exist, and such lesions could be a diagnostic pitfall for gynecologists and pathologists; in these cases, radiologic-pathologic correlation and radiologic guided biopsy are mandatory.

Development of a touchdown droplet digital PCR assay for the detection and quantitation of human papillomavirus 16 and 18 from self-collected anal samples

ABSTRACT Broad-spectrum droplet digital polymerase chain reaction (ddPCR) assays can be used to quantitate multiple human papillomavirus (HPV) types. However, the presence of nucleotide mismatches at the primer binding sites may result in different optimal annealing temperatures and, consequently, under detection of some HPV types. Touchdown PCR uses decreasing annealing temperature over the first PCR cycles allowing for specific and efficient amplification of the target sequence. Here, as a proof-of-concept, we assessed if touchdown ddPCR (TD-ddPCR) using broad-spectrum oligonucleotides and genotype-specific probes targeting HPV L1 can be used to quantitate HPV. We tested our hypothesis using HPV positive cell lines and anal samples which potentially contain PCR inhibitors. Using serial dilutions of HPV16 and HPV18 DNA, the TD-ddPCR assays were performed with linearity across the dilution series (Pearson’s correlation r = 0.9999) and detected one copy of both HPV types, with no cross-reactivity to 27 other low-risk and high-risk HPV types. The PCRs were highly reproducible, with inter- and intra-assay coefficient of variations ranging 2.0–6.3% for HPV16 and 4.9–11.6% for HPV18. Testing self-collected anal samples positive for HPV16 ( n = 29) or HPV18 ( n = 33) and those negative for both HPV types ( n = 22) with the ddPCR assay and Seegene Anyplex II HPV28 (Seegene, TD-ddPCR) showed high agreements between the two assays. In conclusion, HPV broad-spectrum primers containing mismatches at their target binding sites and a touchdown thermal cycling approach resulted in sensitive and specific amplification of HPV genome using ddPCR. The assay has potential clinical diagnostic applications for detection, quantitation, and monitoring of anal HPV infections. IMPORTANCE The quantity of the human papillomavirus (HPV) is associated with disease outcome. We designed an accurate and precise digital PCR assay for quantitating HPV in anal samples, a sample type that is typically problematic due to the presence of PCR inhibitors.

Investigation of human papilloma virus frequency in pregnant women.

The aim of this study was to determine the frequency of human papillomavirus (HPV), HPV types, and the association with age, pregnancy, and childbirth in pregnant women. Four hundred and forty-eight pregnant women who visited our clinic were enrolled in the study. Polymerase chain reaction was used to determine HPV DNA and typing by cervical smear in the initial applications of patients with detailed history. Statistical Package for the Social Sciences (SPSS) 13.0 was used to analyze the data. HPV DNA was positive in 26 (5.8%) of 448 patients who participated in the study. High-risk HPV types were detected in 77% of HPV-positive cases and low-risk HPV types in 23% of cases. Only one HPV type was detected in 13 patients (50%), while multiple HPV types were detected in 13 patients (50%) of the HPV-positive cases. A significant negative association was found between gravidity and positivity of HPV DNA. HPV is more common in women with lower parity and gravidity. Studies conducted in different populations in different regions of Turkey have shown that the incidence of HPV in women ranges from 2.2% to 26%. In our study, the incidence of HPV DNA in pregnant women was 5.8%. It was found that there was an inverse relationship between parity and HPV DNA positivity. The HPV DNA positivity decreased with the number of births.

Novel competitive enzyme-linked immunosorbent assay for the detection of the high-risk Human Papillomavirus 18 E6 oncoprotein.

Cervical cancer represents a global concern with 604,000 new cases and 342,000 deaths reported annually, with the vast majority diagnosed in low income countries. Despite high-risk Human Papillomavirus (HR HPV)-induced cervical cancer has become highly preventable through prophylactic vaccines, screening programs are critical in the control of cervical carcinogenesis in populations with limited access to vaccination and in older generations of women who have already been exposed to HR HPV infection. The surge of HPV molecular tests has provided a more sensitive and accurate diagnostic alternative to cytology screening. Given that HPV DNA testing presents a low positive predicted value, leading to unnecessary treatment, the E6 oncoprotein from HR HPV types arises as a promising diagnostic marker for its overexpression in transformed HPV-positive cancer cells. For these reasons, this study aimed at obtaining monoclonal antibodies (mAbs) against the E6 oncoprotein of one of the most prevalent HR HPV types worldwide, HPV18, in order to develop a highly specific and sensitive indirect competitive ELISA (icELISA). The production of hybridomas secreting HPV18 E6 mAbs was carried out through a combined tolerization and immunization strategy, in order to avoid cross-reactivity with the E6 protein from low-risk HPV types 6 and 11. We selected the 7D2 hybridoma clone, which recognized HPV18 E6 and showed some cross-reactivity against the HR HPV45 E6 oncoprotein. The 7D2 mAb enabled the development of a sensitive, reliable and reproducible icELISA to detect and quantify small amounts of HPV18 E6 biomarker for cervical cancer progression. The present study establishes a valid 7D2-based icELISA that constitutes a promising bioanalytical method for the early detection and quantification of HPV18 E6 oncoprotein in cervical swab samples and cancer prevention.

Human papillomavirus spectrum of HPV-infected women in Nigeria: an analysis by next-generation sequencing and type-specific PCR

BackgroundHuman papillomavirus (HPV) infection and cervical cancer are leading health problems and causes of death in many parts of the world. There are ~ 200 HPV types that can infect humans. This study aims to understand the spectrum of HPV infections in Nigerian women with normal or abnormal cytology.MethodsWe screened cervical samples from 90 women with possible HPV infections collected in two regional hospitals in Nigeria. The first screening was done using next-generation DNA sequencing (NGS), identifying multiple HPV types in many samples. Thereafter, type-specific PCR analysis was used to verify the NGS-identified HPV types in each sample.ResultsNGS analysis of the 90 samples from the Nigerian cohort identified 44 HPV types. The type-specific PCR confirmed 25 HPV types out of the 44 HPV types detected by NGS, and ~ 10 of these types were the most prevalent. The top five prevalent types found in the Nigerian cohort were HPV71 (17%), HPV82 (15%), HPV16 (16%), HPV6 (10%), and HPV20 (7%). Among the PCR-confirmed HPV types, we found 40.98% high-risk HPV types, 27.22% low-risk HPV types, and 31.15% undetermined HPV types. Among these 25 HPV types in Nigeria, only six were included in the current nine-valent HPV vaccine. We also observed strikingly high multiple HPV infections in most patients, with as many as nine HPV types in a few single samples.ConclusionsOur NGS-PCR approach of HPV typing in the Nigerian cohort samples unveiled all possible HPV types currently circulating in Nigerian people. We confirmed 25 HPV types using NGS and PCR, with many samples infected with multiple HPV types. However, only six of these types are part of the nine-valent HPV vaccines indicating the need to develop region-specific selective vaccines.

Sequence analysis of exons 30 and 31 of LAMA3 gene variants and its association with human papillomavirus infection predisposition: no evidence was found.

Human papillomavirus (HPV) is associated with cervical cancer. For the infection to occur, most HPV types depend on interactions with heparan sulfate proteoglycans (HSPGs); however, non-HSPGs receptors are also involved. Laminin 332 is a crucial component of the epidermis's base membrane. It has shown interactions with HPV that suggest its function as a transient viral receptor in the extracellular matrix (ECM). We provide new information about Laminin 332 and HPV by identifying LAMA3 gene allelic variants from exons 30 and 31 and their distribution among women with and without HPV infection. We included 192 cervical cancer scrape samples from two groups of patients, 96 samples from patients with a low-grade squamous intraepithelial lesion (LSIL) and 96 samples from HPV-negative samples without LSIL. Identification of the HPV type was performed using an LCD-Array kit. Exons 30 and 31 of LAMA3 were amplified by PCR and analyzed by Sanger's sequencing. We identified a wide range of HPV types. The most frequent low-risk (lrHPV) HPV types were 6, 42, 44, and 90. For high-risk (hrHPV) HPV were 16, 31, 56, and 66. Only the genetic variant rs1131521 was identified in both groups. However, no significant association was observed between rs1131521 and the study groups. A single silent polymorphism was identified in both groups with similar frequency, whereas no mutations related to increased epithelial friability were identified.

Association of HPV genotype and phylogenic clade with oropharyngeal cancer outcomes.

6058 Background: Human papillomavirus (HPV) is a causative agent of oropharyngeal squamous cell carcinoma (HPVOPC). There are multiple different HPV genotypes based on variations in the nucleotide sequence of the L1 capsid, with HPV16 being most common in HPVOPC (85%). HPV genotypes can be further classified into phylogenic clades, including alpha-9 (A9) and alpha-7 (A7). HPVOPC is associated with a more favorable prognosis than HPV-negative disease, but the impact of specific HPV genotype and phylogenic clade on patient outcomes is not well understood and has profound implications for de-escalation studies in HPVOPC. Methods: A retrospective cohort study was conducted at a large multicenter health system. Patients diagnosed with non-metastatic HPVOPC or unknown primary from 2012-2021 and treated with curative intent were included. Low-risk HPV types (6, 11, etc) and local recurrences were excluded. HPV molecular typing was prospectively performed according to institutional protocol. Cox regressions were used to measure the effect of HPV genotype (16 versus non-16) on overall survival (OS) and event-free survival (EFS). Gender, smoking status, alcohol use, ECOG performance status, T stage, N stage, and subsite (oropharynx versus known primary) were covariates. A secondary survival analysis according to high-risk HPV clade (A9 versus A7) was also performed. Results: The total cohort included 520 patients, the majority of whom were HPV16 (86.9%, n = 452), A9 clade (97.1%, n = 505), oropharynx primary (94.8%, n = 493), male (86.9%, n = 452), white (72.5%, n = 377), non-Hispanic (74.2%, n = 386), non-active smokers (88.3%, n = 459), ECOG 0 (75.6%, n = 393), and N2 stage (65.2%, n = 339). The most common non-16 HPV genotypes were 18 (n = 7), 33 (n = 21), and 35 (n = 29). HPV genotype was not associated with OS (HR = 1.64, 95% CI 0.85-3.16, p= 0.14) or EFS (HR = 1.15, 95% CI 0.64-2.07, p= 0.65). A7 clade was associated with a worse OS compared to A9 in the univariate analysis (HR = 3.43, 95% CI 1.07-10.97, p= 0.04), but not in the multivariate analysis (HR = 1.67, 95% CI 0.45-6.17, p = 0.44). Finally, HPV clade was not associated with EFS (HR = 1.38, 95% CI 0.34-5.60, p= 0.66). Conclusions: While HPV genotype was not associated with survival differences in patients with HPVOPC, HPV A7 clade only predicted worse OS in univariate analysis. Although the total cohort size was large, this analysis was limited by small numbers of subjects with non-16 HPV genotypes and non-A9 clade, which is reflective of the predominance of HPV16 in HPVOPC. Thus, a pooled multicenter collaboration is needed to further address this question. Moreover, future studies should investigate whether outcomes may be related to intratypic HPV16 variants that affect oncoprotein function and/or immunogenicity. Detection of a subgroup of HPVOPC with possible inferior outcomes has important implications for studying treatment de-escalation in HPVOPC.

Association between cervical immune markers and cervical high-risk HPV infections.

e17522 Background: Evidence suggests that human papillomavirus (HPV) infection of the cervix, alters local immune markers. The aim of this study was to comprehensively evaluate the types and concentration of cervical cytokines and chemokines in HPV-negative compared to HPV-positive women in sub-Saharan African. Methods: The study population was 275 women enrolled in the African Collaborative Center for Microbiome and Genomics (ACCME) HPV cohort study. The concentration of 27 chemokines, cytokines and growth factors was quantified from cervical samples, using multiplexed bead-based immunoassays. HPV types were characterized using SPF25/LiPA10. Regression models were used to estimate the association between each immune marker and HPV infection. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for fold-change in immune marker levels divided into quartiles, using the lowest quantile as a reference category. Results: The mean (±SD) age of the women was 41 (±8) years. Some, 49% (134/275) were HPV negative and 51% (141/275) were HPV positive. Of the HPV positive women, 39% (55/141) had low-risk HPV (lrHPV) types and 93% (131/141) had high-risk HPV (hrHPV) types. Compared to HPV negative women, lrHPV-positive women had significantly higher median concentration of IFN-g, IL-2, IL-4, IL-7, IL-10 and IL-13. While hrHPV-positive women had significantly lower median concentration of GMCSF, interleukin (IL)-1RA and MCP-1; and higher median concentration of interferon-g (IFN-g), IL-b, IL-6, IL-7, IL-8, IL-9, IL-10, IL-13, IL-15 and MCP-1b, compared to HPV negative women. Women had significantly higher odds of lrHPV infection if they were in the highest quartile for IL-2, IL-4, IL-5, IL-7, IL-10, IL-12p70 and IL-13. While women had significantly lower odds of hrHPV infection if they were in the highest quartile for IL-IRA (OR 0.16, 95% CI: 0.07, 0.34) or MCP-1 (OR 0.39, 95% CI: 0.18, 0.83), and significantly higher odds of hrHPV infections if they were in the highest quartile for eotaxin, IFN-g, IL-1b, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-13, IL-15, MIP-1a, MIP-1b or TNFa. Conclusions: Consistent with findings from other populations, higher concentration of pro-inflammatory immune markers in the cervix were associated with higher odds of high-risk HPV infections among African women.

Recurrent Respiratory Papilloma of the Larynx

Recurrent respiratory papilloma (RRP) is a common benign neoplasm that most commonly affects the larynx. Caused primarily by human papilloma virus (HPV) types 6 and 11, RRP affects both children and adults; juvenile-onset RRP is thought to be related to HPV exposure during childbirth, whereas adult-onset disease has been hypothesized to be related to sexual contact.1Aaltonen L.M. Rihkanen H. Vaheri A. Human papillomavirus in larynx.Laryngoscope. 2002; 112: 700-707https://doi.org/10.1097/00005537-200204000-00020Crossref PubMed Scopus (83) Google Scholar Symptoms of RRP are typically voice or airway related. Adults tend to experience voice changes, primarily because of impairment of vocal fold vibration in the setting of papilloma along the membranous vocal folds. In children, in whom lesions tend to grow more quickly, airway narrowing may present with stridor in addition to voice changes. Direct or indirect laryngoscopy is standard work-up. Flexible fiberoptic or distal chip scopes may reveal heaped, irregular lesions of the glottis (Figure 1A) or supraglottis, and filters (eg, narrow-band imaging) (Figure 1B) may help to reveal intraepithelial capillary loops. Biopsy remains the criterion standard for diagnosis. Figure 2 illustrates typical findings, including a higher-power image of a cellular papillary frond with viral cytopathic effects and corresponding chromogenic in situ hybridization for low-risk HPV type 6/11.2Venkatesan N.N. Pine H.S. Underbrink M.P. Recurrent respiratory papillomatosis.Otolaryngol Clin North Am. 2012; 45 (671-94, viii-ix. https://doi.org/10.1016/j.otc.2012.03.006)Abstract Full Text Full Text PDF Scopus (89) Google Scholar Treatment includes excision or ablation with cold technique, photoangiolytic lasers, or microdebriders, with maximal conservation of the subepithelial vocal fold tissue. Recurrent respiratory papilloma routinely recurs, and so adjunct treatments with cidofovir and bevacizumab are advocated for recalcitrant cases.3Pogoda L. Ziylan F. Smeeing D.P.J. Dikkers F.G. Rinkel R.N.P.M. Bevacizumab as treatment option for recurrent respiratory papillomatosis: a systematic review.Eur Arch Otorhinolaryngol. 2022; 279: 4229-4240https://doi.org/10.1007/s00405-022-07388-6Crossref Scopus (3) Google Scholar Emerging evidence suggests that HPV vaccination, even in those already affected, may be beneficial in reducing the severity of disease.4Ponduri A, Azmy MC, Axler E, et al. The efficacy of human papillomavirus vaccination as an adjuvant therapy in recurrent respiratory papillomatosis. Laryngoscope. Published online January 18, 2023. https://doi.org/10.1002/lary.30560.Google Scholar The authors report no competing interests.

Integrative analysis of gene expression and DNA methylation to identify biomarkers of non-genital warts induced by low-risk human papillomaviruses infection

BackgroundHuman papillomaviruses have been shown to dysregulate the gene expression and DNA methylation profiles of their host cells over the course of infection. However, there is a lack of information on the impact of low-risk HPV infection and wart formation on host cell's expression and methylation patterns. Therefore, the objective of this study is to analyse the genome and methylome of common warts using an integrative approach. MethodsIn the present study, gene expression (GSE136347) and methylation (GSE213888) datasets of common warts were obtained from the GEO database. Identification of the differentially expressed and differentially methylated genes was carried out using the RnBeads R package and the edgeR Bioconductor package. Next, functional annotation of the identified genes was obtained using the Database for Annotation, Visualization, and Integrated Discovery (DAVID). Network construction and analyses of the gene-gene, protein-protein, and signaling interactions of the differentially expressed and differentially methylated genes was performed using the GeneMANIA web interface, the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) database, and the Signaling Network Open Resource 2.0 (SIGNOR 2.0), respectively. Lastly, significant hub genes were identified using the Cytoscape application CytoHubba. ResultsA total of 276 genes were identified as differentially expressed and differentially methylated in common warts, with 52% being upregulated and hypermethylated. Functional enrichment analysis identified extracellular components as the most enriched annotations, while network analyses identified ELN, ITGB1, TIMP1, MMP2, LGALS3, COL1A1 and ANPEP as significant hub genes. ConclusionsTo the best knowledge of the authors, this is the first integrative study to be carried out on non-genital warts induced by low-risk HPV types. Future studies are required to re-validate the findings in larger populations using alternative approaches.