Papillomavirus Type Research Articles

Two Lineages of Papillomaviruses Identified from Caracals (Caracal caracal) in South Africa.

Papillomaviruses (PV) infect epithelial cells and can cause hyperplastic or neoplastic lesions. In felids, most described PVs are from domestic cats (Felis catus; n = 7 types), with one type identified in each of the five wild felid species studied to date (Panthera uncia, Puma concolor, Leopardus wiedii, Panthera leo persica and Lynx rufus). PVs from domestic cats are highly diverse and are currently classified into three genera (Lambdapapillomavirus, Dyothetapapillomavirus, and Taupapillomavirus), whereas those from wild felids, although diverse, are all classified into the Lambdapapillomavirus genus. In this study, we used a metagenomic approach to identify ten novel PV genomes from rectal swabs of five deceased caracals (Caracal caracal) living in the greater Cape Town area, South Africa. These are the first PVs to be described from caracals, and represent six new PV types, i.e., Caracal caracal papillomavirus (CcarPV) 1-6. These CcarPV fall into two phylogenetically distinct genera: Lambdapapillomavirus, and Treisetapapillomavirus. Two or more PV types were identified in a single individual for three of the five caracals, and four caracals shared at least one of the same PV types with another caracal. This study broadens our understanding of wild felid PVs and provides evidence that there may be several wild felid PV lineages.

Diverse papillomaviruses identified from Antarctic fur seals, leopard seals and Weddell seals from the Antarctic

Papillomaviruses (family Papillomaviridae) are non-enveloped, circular, double-stranded DNA viruses known to infect squamous and mucosal epithelial cells. In the family Papillomaviridae there are 53 genera and 133 viral species whose members infect a variety of mammalian, avian, reptilian, and fish species. Within the Antarctic context, papillomaviruses (PVs) have been identified in Adélie penguins (Pygoscelis adeliae, 2 PVs), Weddell seals (Leptonychotes weddellii, 7 PVs), and emerald notothen (Trematomus bernacchii, 1 PV) in McMurdo Sound and Ross Island in eastern Antarctica. Here we identified 13 diverse PVs from buccal swabs of Antarctic fur seals (Arctocephalus gazella, 2 PVs) and leopard seal (Hydrurga leptonyx, 3 PVs) in western Antarctica (Antarctic Peninsula), and vaginal and nasal swabs of Weddell seals (8 PVs) in McMurdo Sound. These PV genomes group into four genera representing 11 new papillomavirus types, of which five are from two Antarctic fur seals and a leopard seal and six from Weddell seals.

Učinci ekstrakta Cytinus hypocistis (L.) L. u životinjskom modelu neoplazije inducirane papiloma virusom

Infections with certain types of papillomavirus, such as the human papillomavirus 16 (HPV16), are associated with the development of preneoplastic lesions and cancers of the anogenital, and head and neck regions. Cytinus hypocistis (L.) L. extracts are composed of substances presenting antiproliferative, antioxidant, anti-inflammatory and antibacterial properties, which might be promising as new therapeuticcompounds. This study analysed the influence of topical application of an extract obtained from C. hypocistis (CH) on K14-HPV16 and FVB/n mice to evaluate its therapeutic and toxicological properties. To achieve the study goals,30 female mice, 33–37 weeks old, were divided into six groups (n=5/group): I (HPV+CH3.1); II (HPV+CH6.2); III (HPV+CH12.4); IV (FVB/n+CH12.4); V (HPV+control) and VI (FVB/n+-control). CH was applied topically to both ears for 28 days. After this period, all animals were sacrificed for samples collection. Skin lesions were classified histologically. Toxicological parameters included haematological and biochemical blood markers, and hepatic oxidative stress analysis. Transgenic animals showed a decrease in mean body weight regardless of the concentration of extract applied. The extract had no influence on physiological parameters, organ weight, or biochemical and oxidative stress parameters. Histology demonstrated the presence of proliferative epithelial lesions in the skin and oral mucosa of K14-HPV16 mice, with no association with the application of this extract. Overall, the application of CH extract had no influence on the skin lesions and was well tolerated by the animals in these concentrations.

Characterization of a Chimeric MmuPV1 Genome with HPV-16E6E7

Background/Objectives: Papilloma Viruses (PVs) are double-stranded DNA viruses that infect cutaneous and mucosal epithelium. In humans, there are over 100 types of PVs. HPV-16 is a high-risk type that causes ~50% of cervical cancers and ~70% of oropharyngeal cancers by expressing viral oncogenes E6 and E7 in replicating keratinocytes. The mouse PV, MmuPV1, displays species tropism and causes neoplastic lesions. Its discovery in 2011 introduced the opportunity to study PVs from early stages of infection to cancer development. Such an in vivo model of HPV-16 could uncover novel mechanisms for treatment intervention and disease prevention. Our study aims to characterize a small-animal infection model using a chimeric MmuPV1 genome with HPV-16 E6 and E7 in place of mouse E6 and E7 (MmuPV1-16E6E7). The goal of our study is to investigate the genome’s ability to express the HPV-16 oncogenes in vitro and to cause tumors in vivo. Methods: To increase selective pressure in vitro, we replaced the MmuPV1 L1 and L2 genes with a neomycin cassette (MmuPV1-16E6E7neo). We packaged these genomes into infectious quasiviruses using a HEK 293TTF viral packing cell line. After isolating quasiviruses, we infected two donor human foreskin keratinocytes (HFK) cell lines. Forty-eight hours post infection, we isolated mRNA for qPCR to quantify HPV-16 E6 expression. To investigate whether MmuPV1-16E6E7 can cause tumors in in vivo, we infected athymic nude (N/J) mice orally with quasiviruses to evaluate infection and viable genome persistence. Results: HPV-16 E6 cDNA was detected in three HFK isolates. Conclusions/Future Directions: The presence of HPV-16 E6 cDNA indicates that the gene can be successfully transcribed from MmuPV1-16E6E7neo in HFKs. Future studies will assess this ability in primary mouse keratinocytes. If tumors are observed, we can use this model to study the efficacy of antiviral compounds to inhibit HPV16 E6 in vivo.

Acral vasculitis and Chilblain-like lesions: is it COVID-19 infection?

The cutaneous manifestations associated with COVID-19 are increasingly reported, their exact incidence remains to be estimated, their pathophysiological mechanisms are largely unknown and the role, direct or indirect, of SARS-CoV-2 in their pathogenesis is still debated. We present a case of COVID-19 that manifested with a purpuric vasculitis pattern on the lower extremities and chilblain-like lesion in toe in a patient presented COVID-19 lungs involvement with polymerase chain reaction and serology are negatives. The reported presenting dermatological symptoms: associated chilblain-like lesions and vasculitis pattern of patient with negative COVID-19 RT-PCR and serology. The polymorphism of cutaneous lesions associated with the COVID-19 infection can mimic or appear similar to other well-known conditions. Even though, RT- PCR and serology are negatives, an infection of SARS-CoV-2 should be considerate.

The effectiveness of the application of modern diagnostic criteria of cervical pathology

BACKGROUND: Cervical cancer is one of the four most common cancers today and is the second most common cancer among women in developing countries. According to World Health Organization estimates, in 2018, the number of new cases in these countries was 570,000 (84% of all new cases globally). About 311,000 women died of cervical cancer in 2018, with more than 85% of these deaths occurring in low- and middle-income countries. The detection of the disease in its later stages has not significantly reduced since 2018 (32.6% of all cases).
 AIM: The aim of this study was to compare the information contents of conventional and liquid cytology methods in women with verified human papillomavirus of high oncogenic risk.
 MATERIALS AND METHODS: We retrospectively analyzed 50 outpatient records of women aged 23 to 65 years who underwent colposcopy (cervical biopsy) based on the results of cervical screening. To establish the diagnosis, all patients underwent examination of smears from the cervix and cervical canal using conventional (n = 25) and liquid (n = 25) cytology methods. A molecular genetic study was carried out to detect high-oncogenic human papillomavirus types (16, 18, 31, 33, 35, 45, 51, 56, 58) in scraping of the cervix by real-time polymerase chain reaction. Women in the both study groups also underwent colposcopy with histological examination of the cervical biopsy specimen. The analysis of the obtained data was carried out using methods of parametric and nonparametric statistics. Inclusion criteria included cytological examination, verified human papillomavirus of high oncogenic risk, and colposcopy. Exclusion criteria included pregnancy and acute infectious diseases.
 RESULTS: According to the results of histological examination, the percentage of coincidence of cytological and histological diagnosis in the group of women who underwent conventional cytological examination was 48%. The total number of discrepancies in this group was 13 (52.0 9.99%). The number of women infected with human papillomavirus in the both study groups was almost the same (18 in the conventional cytology group and 19 in the liquid cytology group). The most common types of human papillomavirus among the examined patients were the 16th type (9 women) and the 18th type (6 women).
 CONCLUSIONS: To date, cytological research methods have not lost their relevance, the liquid cytology method being a reliable screening test with high specificity, which reduces the number of unsatisfactory preparations for analysis and false negative results. The most informative for women with type 2 or type 3 of transformation zone is the use of the liquid cytology method.

Association of abnormal vaginal microflora and HPV infection in cervical precancerous lesions: a retrospective study

The aim of this study was to analyze the correlation between abnormal vaginal microflora and different types of human papillomavirus (HPV) infection and cervical precancerous lesions during the perimenopausal period. This retrospective study included women patients who underwent liquid-based cytologic test (LBC), HPV test, leucorrhea routine test, or routine urine test at the China-Japan Friendship Hospital between October 2019 and January 2020. A cut-off of 45 years was used as the cut-off age for menopause. The positivity and subtypes of HPV were determined using a chip-based assay. Vaginal microflora was determined using an HB-2012a flow-through hybridization instrument. A total of 132 patients were included in this study. 97 patients were younger than 45 years of age, with a median age of 35 (8.0), and 35 patients ≥ 45 years of age, with a median age of 55 (11.0). There were no significant differences in cytology, type of cervical lesion, HPV type, and common pathogens of the reproductive tract (all p > 0.05). The multivariable analysis showed that only HPV-16 infection lesions (OR: 2.825, 95% CI: 1.121-7.120, p = 0.028), Chlamydia trachomatis infection (OR: 0.142, 95% CI: 0.024-0.855, p =0.033), and Mycoplasma infection (OR: 7.750, 95% CI: 1.603-37.474, p = 0.011) were independent risk factors for cervical precancerous lesions. The menopausal status (with age < 45 or > 45 years as its surrogate) was not associated with cervical precancerous lesions. Menopause was not associated with cervical precancerous lesions. The results suggest that the prevention and treatment of HPV-16, Chlamydia trachomatis infection, and Mycoplasma infection could be significant to prevent the occurrence of cervical precancerous lesions.

Awareness of the link between HPV, cervical cancer, and HPV vaccination: An online survey among Polish women.

10538 Background: In 2019 cervical cancer (CC) was the 7th most common malignant neoplasm in Polish women. Its incidence has been declining over the years, nonetheless the 5-year survival rate is still significantly lower than the EU average. Vaccination against human papillomavirus (HPV) is not included in the mandatory immunization schedule in Poland and is not reimbursed yet. The aim of the study was to assess women's awareness of the risk factors of CC (including the role of HPV) and the possibility of cancer prevention through HPV vaccination. Methods: The questionnaire prepared by the study team was aimed at women and conducted online using social network sites. The single/multiple-choice questions included knowledge about CC, HPV and HPV vaccination. Statistical analyses were performed using MS Excel 2016 and Medistica pvalue.io 2021 software in general population and age- dependent subgroups. Results: The total number of 3262 women participated in the study. The age subgroups were: aged ≤ 18 years (group 1): 1090 (33.4%), aged 19-25 years (group 2): 1615 (49.5%) and &gt; 25 years (group 3): 557 (17.1%). Overall 81.4% of the respondents correctly associated HPV as the key factor of CC, 85.0% believed that CC could be prevented and 80.0% had heard of HPV vaccination before. In the last two parameters a statistically significant difference in age subgroups was observed (p &lt; 0.001). Only 21.2% of the total number of respondents were vaccinated (26.2%, 22.0% and 9.2% in age groups 1, 2 and 3 respectively, p &lt; 0.001). In all studied populations the following factors had the greatest influence on the decision to get vaccinated: parents’ advice – 54.8%, pediatrician recommendation – 15.7%, possibility of free vaccination offered by municipality – 11.6%, teachers’ advice - 9.7%, gynecologist recommendation - 8.2%. The main reasons behind not getting vaccinated were: high price (63.0%) and disbelief in efficacy (41.5%); only 12.2% respondents were afraid of possible side effects. 65.0% of the unvaccinated respondents declared willingness to get vaccinated in the future (71.5%, 66.1% and 52.0% in age groups 1, 2 and 3 respectively p &lt; 0.001), 28.5% of the respondents were not confident of their decision at the time of submitting answers. Conclusions: As oncogenic types of human papillomavirus play a vital role in pathogenesis of CC, the HPV vaccination and cytological screening together with awareness-raising campaigns should be considered the best strategy for the prevention of CC, its early detection and increase in survival rates. Vaccine willingness remains at high level especially among young women, however the effectiveness of vaccination should be emphasized and the HPV free vaccination programs should be introduced by the state. The educational role of gynecologists needs to be enhanced.

A phase II study of pembrolizumab for HPV-associated papilloma patients with laryngeal, tracheal, and/or pulmonary involvement.

2590 Background: Recurrent respiratory papillomatosis (RRP) is caused by human papillomavirus (HPV) types 6 &amp; 11. RRP proliferates in the squamous epithelium lining the respiratory tract impacting breathing, swallowing, and voice and carries a 3-5% risk of malignant transformation. Given the multi-focality of the disease and tolerized host immune response against HPV, in part through upregulation of the PD1:PDL1 axis, the safety and efficacy of systemic pembrolizumab (pembro) as a novel treatment for this benign tumor patient (pt) population was evaluated in a phase II clinical trial. Methods: RRP pts &gt; 12 years of age were treated with pembro 200mg every 3 weeks. Primary endpoints were best overall response (ORR) (measured by endoscopic lesional burden) and safety. Greater than 5 pts with disease response out of 21 (assuming &gt; 1 of first n = 11 with disease in response) provided 86% power to distinguish between a 15% and a 38% ORR (one-sided 8% binomial test). HPV-specific CD8+ T cell frequency and functional states and biomarkers of response and immune resistance are being evaluated in serial tissue and liquid biopsies (up to 8 biopsies/patient over the 24 months of treatment). Results: The Simon two-stage, stage 1 criteria was met. A total of 21 patients were enrolled and all are now off treatment. Median age (range) was 45 (19-68), 57% (12/21) were male and 67% (14/21) were white. 48% (10/21) had Juvenile-onset (Jo)-RRP, 57% (12/21) had pulmonary RRP involvement, and 19% (4/21) had SCC derived from their RRP. 62% (13/21) completed 24 months of treatment. Reasons for discontinuation included disease progression (14%, 3/21), treatment related adverse event (TRAEs) (14%, 3/21), and study withdrawal (10%, 2/21). A partial response (≥25% reduction in endoscopic tumor burden score) was observed in 57% (12/21) (95% CI: 34%-78.2%) of pts (7 of 10 with Jo-RRP and 5 of 11 with Adult-onset (Ao)-RRP disease responded). Stable disease was observed in 33% (7/21). No complete responses were observed. Fatigue was the most frequent TRAEs; Grade 3 TRAEs included uveitis and hypophysitis, both of which were reversible upon pembro discontinuation and steroid use. At a median follow-up: 25.6 (6.2-38.1 months), the mean number of surgical interventions was reduced by 7 surgeries/year (p = 0.004) in pts treated on the trial for &gt; 12 months, and, upon treatment completion, durable clinical benefit was observed with no additional treatment needed for the duration of the clinical trial follow-up for some pts. Conclusions: Pembro reduces the need for routine surgical interventions based on the durable response rates being achieved. Further study of pembro +/- other agents is warranted to achieve and sustain complete responses in this population. Clinical trial information: NCT02632344.

The role of viral infection in the pathology of oral mucosa in persons with prosthetic constructions

Aim.To detect viral infection of the oral mucous membrane in individuals with dentures.Material and methods.110 patients with removable and 100 - with fixed prostheses were examined. Diagnostics of virus infection were carried out by ELISA and PCR methods (Kvant-21).Results and discussion.It has been revealed that patients with removable prostheses revealed oral mucosa diseases in 91 (82,7%) persons and with fixed prostheses - in 75 (75,0%) patients. Viral infection in patients with removable dentures is detected in 58 (63.7%;91) and with fixed dentures in 43 (57.3%;75) patients. Among patients with removable dentures, HPV types 11 and 16 occur 1.3 times more frequently than in patients with fixed dentures, and HPV type 6 - 4.6 times less frequently. HPV types 11 and 16 are most frequently detected in patients with removable prostheses in the case of red lichen planus while in the group of patients with fixed prostheses they are more frequently observed in patients suffering from periodontal disease.Conclusion.Viral infection plays an important role in the development of diseases of the oral mucosa, which differ depending on the presence of removable and fixed dental structures. Papillomavirus types 6, 11, and 16 have the most pronounced effect on the course of oral mucosa diseases.

Non-human primate papillomavirus E6-mediated p53 degradation reveals ancient evolutionary adaptation of carcinogenic phenotype to host niche.

Non-human primates (NHPs) are infected with papillomaviruses (PVs) closely related to their human counterparts, but there are few studies on the carcinogenicity of NHP-PVs. Using an in vitro cell co-transfection assay, we systematically screened the biochemical activity of E6 proteins encoded by macaque PVs for their ability to bind and promote degradation of host p53 proteins. A host species barrier exists between HPV16 and MfPV3 with respect to E6-mediated p53 degradation that is reversed when p53 residue 129 is swapped between human and macaque hosts. Systematic investigation found that E6 proteins encoded by most macaque PV types in the high-risk species α12, but not other Alpha-PV clades or Beta-/Gamma-PV genera, can effectively promote monkey p53 degradation. Interestingly, two macaque PV types (MfPV10 and MmPV1) can simultaneously inhibit the expression of human and monkey p53 proteins, revealing complex cross-host interactions between PV oncogenes and host proteomes. Single point-mutant experiments revealed that E6 residue 47 directly interacts with p53 residue 129 for host-specific degradation. These findings suggest an ancient host niche adaptation toward a carcinogenic phenotype in high-risk primate PV ancestors. Following periods of primate host speciation, a loss-of-function mutation model could be responsible for the formation of a host species barrier to E6-mediated p53 degradation between HPVs and NHP-PVs. Our work lays a genetic and functional basis for PV carcinogenicity, which provides important insights into the origin and evolution of specific pathogens in host pathogenesis.

Tumor Cell Plasticity in Equine Papillomavirus-Positive Versus-Negative Squamous Cell Carcinoma of the Head and Neck.

Squamous cell carcinoma of the head and neck (HNSCC) is a common malignant tumor in humans and animals. In humans, papillomavirus (PV)-induced HNSCCs have a better prognosis than papillomavirus-unrelated HNSCCs. The ability of tumor cells to switch from epithelial to mesenchymal, endothelial, or therapy-resistant stem-cell-like phenotypes promotes disease progression and metastasis. In equine HNSCC, PV-association and tumor cell phenotype switching are poorly understood. We screened 49 equine HNSCCs for equine PV (EcPV) type 2, 3 and 5 infection. Subsequently, PV-positive versus -negative lesions were analyzed for expression of selected epithelial (keratins, β-catenin), mesenchymal (vimentin), endothelial (COX-2), and stem-cell markers (CD271, CD44) by immunohistochemistry (IHC) and immunofluorescence (IF; keratins/vimentin, CD44/CD271 double-staining) to address tumor cell plasticity in relation to PV infection. Only EcPV2 PCR scored positive for 11/49 equine HNSCCs. IHC and IF from 11 EcPV2-positive and 11 EcPV2-negative tumors revealed epithelial-to-mesenchymal transition events, with vimentin-positive cells ranging between <10 and >50%. CD44- and CD271-staining disclosed the intralesional presence of infiltrative tumor cell fronts and double-positive tumor cell subsets independently of the PV infection status. Our findings are indicative of (partial) epithelial–mesenchymal transition events giving rise to hybrid epithelial/mesenchymal and stem-cell-like tumor cell phenotypes in equine HNSCCs and suggest CD44 and CD271 as potential malignancy markers that merit to be further explored in the horse.

Seroprevalence of mucosal and cutaneous human papillomavirus (HPV) types among children and adolescents in the general population in Germany

BackgroundIn Germany, HPV vaccination of adolescent girls was introduced in 2007. Nationally representative data on the distribution of vaccine-relevant HPV types in the pre-vaccination era are, however, only available for the adult population. To obtain data in children and adolescents, we assessed the prevalence and determinants of serological response to 16 different HPV types in a representative sample of 12,257 boys and girls aged 1–17 years living in Germany in 2003–2005.MethodsSerum samples were tested for antibodies to nine mucosal and seven cutaneous HPV types. The samples had been collected during the nationally representative German Health Interview and Examination Survey for Children and Adolescents in 2003–2006. We calculated age- and gender-specific HPV seroprevalence. We used multivariable regression models to identify associations between demographic and behavioral characteristics and HPV seropositivity.ResultsWe found low but non-zero seroprevalence for the majority of tested HPV types among children and adolescents in Germany. The overall seroprevalence of HPV-16 was 2.6%, with slightly higher values in adolescents. Seroprevalence of all mucosal types but HPV-6 ranged from 0.6% for HPV-33, to 6.4% for HPV-31 and did not differ by gender. We found high overall seroprevalence for HPV-6 with 24.8%. Cutaneous HPV type seroprevalence ranged from 4.0% for HPV-38 to 31.7% for HPV-1. In the majority of cutaneous types, seroprevalence did not differ between boys and girls, but increased sharply with age, (e.g., HPV-1 from 1.5% in 1–3-years-old to 45.1% in 10–11-years-old). Associations between behavioral factors and type-specific HPV prevalence were determined to be heterogeneous.ConclusionsWe report the first nationally representative data of naturally acquired HPV antibody reactivity in the pre-HPV-vaccination era among children and adolescents living in Germany. These data can be used as baseline estimates for evaluating the impact of the current HPV vaccination strategy targeting 9–14-years-old boys and girls.

New approach for genomic characterisation of equine sarcoid-derived BPV-1/-2 using nanopore-based sequencing

BackgroundBovine papillomavirus (BPV) types 1 and 2 play a central role in the etiology of the most common neoplasm in horses, the equine sarcoid. The unknown mechanism behind the unique variety in clinical presentation on the one hand and the host dependent clinical outcome of BPV-1 infection on the other hand indicate the involvement of additional factors. Earlier studies have reported the potential functional significance of intratypic sequence variants, along with the existence of sarcoid-sourced BPV variants. Therefore, intratypic sequence variation seems to be an important emerging viral factor. This study aimed to give a broad insight in sarcoid-sourced BPV variation and explore its potential association with disease presentation.MethodsIn order to do this, a nanopore sequencing approach was successfully optimized for screening a wide spectrum of clinical samples. Specimens of each tumour were initially screened for BPV-1/-2 by quantitative real-time PCR. A custom-designed primer set was used on BPV-positive samples to amplify the complete viral genome in two multiplex PCR reactions, resulting in a set of overlapping amplicons. For phylogenetic analysis, separate alignments were made of all available complete genome sequences for BPV-1/-2. The resulting alignments were used to infer Bayesian phylogenetic trees.ResultsWe found substantial genetic variation among sarcoid-derived BPV-1, although this variation could not be linked to disease severity. Several of the BPV-1 genomes had multiple major deletions. Remarkably, the majority of them cluster within the region coding for late viral genes. Together with the extensiveness (up to 603 nucleotides) of the described deletions, this suggests an altered function of L1/L2 in disease pathogenesis.ConclusionsBy generating a significant amount of complete-length BPV genomes, we succeeded to introduce next-generation sequencing into veterinary research focusing on the equine sarcoid, thus facilitating the first report of both nanopore-based sequencing of complete sarcoid-sourced BPV-1/-2 and the simultaneous nanopore sequencing of multiple complete genomes originating from a single clinical sample.

Učestalost infekcije visoko-rizičnim tipovima humanih papiloma virusa na grliću materice žena reproduktivnog perioda

Introduction: Human papilloma virus is the most important etiological factor that leads to the malignant alteration of cervical epithelium. More than 200 types of human papillomavirus are identified. Based on its oncogenic potential, human papillomavirus can be classified as low-oncogenic and high-oncogenic types. Aim: To determine the frequency of infection with high-risk human papillomaviruses in the cervical swabs of reproductively active women in our population. Material and methods: The research is designed as descriptive study. Data on the presence of HPV infections, as well as the virus genotype in 707 cervical swabs from the period from January 2019 to August 2020, together with the correlation with the woman's age and cytological status on the cervix, were analyzed. Results: Human papilloma virus infection was present in 223 out of 707 tested samples (31.54%). The highest frequency of HPV infection, as well as the highest number of HPV genotypes was in the group of women aged 25 - 34 years (36.32%). The frequency of HPV infection correlates with altered cytological status. The presence of 26 HPV genotypes was determined, 16 highly oncogenic and 10 low oncogenic types, of which the most common was highly oncogenic type 16 (25.11%). Conclusion: These findings showed a high frequency of highly oncogenic HPV genotypes, primarily type 16 in most groups of women of reproductive age. In addition, the frequency of infection correlates with cytological status, which indicates the great importance of prevention of genital HPV infection in our female population.

Abstract P043: A double-blind randomized, placebo-controlled trial of oral administration with human papillomavirus (HPV) type 16 E7-expressing Lactobacillus-based vaccine, BLS-ILB-E710c, for the treatment of cervical intraepithelial neoplasia (CIN2/3)

Abstract Background - Despite preventive HPV vaccines is implemented worldwide, current treatments for high grade cervical intraepithelial neoplasia are ablative, and no pharmacological treatments are available. Here we evaluated safety of BLS-ILB-E710c, HPV 16 E7-expressing Lactobacillus-based vaccine, and explored its efficacy for histopathological regression in women with CIN2/3. Methods - Safety and efficacy of BLS-ILB-E710c were assessed in CIN2/3 associated with HPV16 and its related HPV types in a randomized, double-blind, placebo-controlled phase 2b study. Total of 116 patients was recruited from fourteen hospitals in South Korea, and were randomized (2:1) to receive 1000 mg BLS-ILB-E710c or placebo for 5 days at 1, 2, 4, and 8 weeks. The primary endpoint was histopathological regression to CIN1 or normal pathology at 16 weeks after the first dose. Full analysis set (FAS) and Per-protocol set (PPS) analyses were based on patients receiving at least one oral vaccination, and on patients receiving four rounds of oral vaccination without protocol violations, respectively. The safety population included all patients who enrolled. The trial is registered at clinicaltrials.gov (number NCT03274206). Findings - Total of 116 patients were randomized, and 113 received either BLS-ILB-E710c (n=75) or placebo (n=38). The oral vaccination was well tolerated and no serious vaccine-related AEs occurred. No differences were showed between the BLS-ILB-E710c and placebo groups for patient background and adverse events. In the full analysis set (FAS) no statistically significant difference was noted between the two groups of histopathological regression at 16 weeks. However, the distribution by Bethesda system (for cervical cytology) in CIN2 patients showed significant differences between two groups (P=0.0304), which can affect histopathological regression. Sub-group analysis in FAS is performed to reduce the bias; histopathological classification (CIN2, CIN3) and cytological classification (≤ASCUS, LSIL, HISL). Therefore, in CIN3 sub-group, BLS-ILB-E710c recipients showed 13.33% of higher histopathological regression at 32 weeks than placebo recipients, and interestingly represented the statistically significant difference when histopathological regression at 32 weeks compared with 16 weeks (percentage point difference 34.87 [95% CI 7.78–61.96]; P=0.0216). In BLS-ILB-E710c recipients of CIN3 sub-group with histopathological regression, E7-specific CD8+ T lymphocyte immune responses were induced at 32 weeks (P= 0.0323). In addition, in HSIL sub-group, BLS-ILB-E710c recipients showed higher histopathological regression at 32 weeks than placebo recipients in similar with CIN3 sub-group analysis. Interpretation – Based on the finding this study, BLS-ILB-E710c could induce the local immune response by recruiting HPV16 E7-specific CD8+ T cells to cervical lesions, leading to histologic regression. To assess the efficacy of BLS-ILB-E710c as a novel Lactobacillus-based oral vaccine, it is necessary to define the population and time frame for phase 3 trial. Citation Format: Jae Kwan Lee, Seung Hun Song, Young Tae Kim, Chi-Heum Cho, Chan Joo Kim, Young-Chul Park. A double-blind randomized, placebo-controlled trial of oral administration with human papillomavirus (HPV) type 16 E7-expressing Lactobacillus-based vaccine, BLS-ILB-E710c, for the treatment of cervical intraepithelial neoplasia (CIN2/3) [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2021 Oct 7-10. Philadelphia (PA): AACR; Mol Cancer Ther 2021;20(12 Suppl):Abstract nr P043.

Viral load as a factor of persistence of papillomavirus infection in cervical intraepithelial neoplasia grade I

Introduction. The viral antigenic load can influence the nature of the infectious response, leading to elimination of the virus or to chronicity of the process, and in some cases to the progressive course of the disease. The aim of the study was to investigate the relationship between the types of viral load of human papillomavirus and the age of a patient with cervical cervical intraepithelial neoplasia grade I. Material and methods. 86 patients with cervical papillomavirus-induced cervical intraepithelial neoplasia grade I by liquid oncocytology were examined. The frequency of human papillomavirus genotypes of high carcinogenic risk in this group of women was analyzed taking into account the viral load by polymerase chain reaction with real-time hybridization-fluorescence detection. The correlation between the viral load of the human papillomavirus and the age of the patient with further prediction of the regression or progression of cervical pathology was studied. Results. This study found that female patients under 30 years of age with cervical intraepithelial neoplasia grade I associated with papillomavirus infection showed a decrease in viral load after 12 months of follow-up in 30.2%, and in patients with cervical intraepithelial neoplasia grade I of older age — in 9.3%. Discussion. The management tactics for grade I cervical intraepithelial neoplasia associated with papillomavirus infection depends largely on the age of the patients and the degree of viral load. In young patients with immediate reproductive plans, a wait-and-see approach with dynamic observation for 12 months without drug therapy is possible in view of the probable spontaneous elimination of the human papillomavirus within a year. The results of the study suggest a higher rate of human papillomavirus elimination in cervical intraepithelial neoplasia grade I in women under 30 years of age. Conclusion. In cervical intraepithelial neoplasia of degree I in women under 40, infection with several types of the human papilloma virus (16, 18, 31, 51) with a clinically significant viral load (over 4.69±0.07 HPV per 100,000 cells) was found most frequently. In the group of the older reproductive age (40-49 years old), HPV HCC is less frequent in 6.90%. And the predominant role belongs to type 16, which was spread in 69.7% of cases.

The Development of Novel Primer Sets to Specifically Amplify Each of the Five Different Deltapapillomaviruses That Cause Neoplasia after Cross-Species Infection.

Bovine papillomavirus (BPV) types 1 and 2 are recognized as the main cause of equine sarcoids. However, some studies report that up to a quarter of these tumors do not contain detectible BPV1 or BPV2 DNA. The absence of detectible BPV1 or BPV2 in these sarcoids suggests the possible involvement of other papillomavirus types. Currently, five deltapapillomaviruses are recognized to cause mesenchymal neoplasia after cross-species infection. In addition to BPV1 and BPV2, BPV13 has been associated with equine sarcoids in Brazil, BPV14 has been associated with feline sarcoids, and Ovis aries papillomavirus 2 caused a sarcoid-like lesion in a pig. To investigate the cause of equine sarcoids, PCR primers were developed to specifically amplify each of the five different deltapapillomaviruses that have been associated with mesenchymal neoplasia. The specificity of these primers was confirmed using samples of formalin-fixed tissue known to contain each PV type. These primers allow rapid and sensitive detection of deltapapillomavirus DNA in equine sarcoids. As studies have revealed marked regional variability in the cause of equine sarcoids, these primers will be useful to determine the predominant PV type causing sarcoids in a region. Additionally, there is a single report describing mixed infections by BPV1 and BPV2 in equine sarcoids. The specific primer sets are expected to enable more sensitive detection of mixed infections in equine sarcoids. Determining the cause of equine sarcoids is important as vaccines are developed to prevent these common malignant neoplasms.

K-Mer Analyses Reveal Different Evolutionary Histories of Alpha, Beta, and Gamma Papillomaviruses.

Papillomaviruses (PVs) are a heterogeneous group of DNA viruses that can infect fish, birds, reptiles, and mammals. PVs infecting humans (HPVs) phylogenetically cluster into five genera (Alpha-, Beta-, Gamma-, Mu- and Nu-PV), with differences in tissue tropism and carcinogenicity. The evolutionary features associated with the divergence of Papillomaviridae are not well understood. Using a combination of k-mer distributions, genetic metrics, and phylogenetic algorithms, we sought to evaluate the characteristics and differences of Alpha-, Beta- and Gamma-PVs constituting the majority of HPV genomes. A total of 640 PVs including 442 HPV types, 27 non-human primate PV types, and 171 non-primate animal PV types were evaluated. Our analyses revealed the highest genetic diversity amongst Gamma-PVs compared to the Alpha and Beta PVs, suggesting reduced selective pressures on Gamma-PVs. Using a sequence alignment-free trimer (k = 3) phylogeny algorithm, we reconstructed a phylogeny that grouped most HPV types into a monophyletic clade that was further split into three branches similar to alignment-based classifications. Interestingly, a subset of low-risk Alpha HPVs (the species Alpha-2, 3, 4, and 14) split from other HPVs and were clustered with non-human primate PVs. Surprisingly, the trimer-constructed phylogeny grouped the Gamma-6 species types originally isolated from the cervicovaginal region with the main Alpha-HPV clade. These data indicate that characterization of papillomavirus heterogeneity via orthogonal approaches reveals novel insights into the biological understanding of HPV genomes.

Papillomaviruses in Domestic Cats.

Papillomaviruses (PVs) are well established to cause hyperplastic papillomas (warts) in humans and animals. In addition, due to their ability to alter cell regulation, PVs are also recognized to cause approximately 5% of human cancers and these viruses have been associated with neoplasia in a number of animal species. In contrast to other domestic species, cats have traditionally been thought to less frequently develop disease due to PV infection. However, in the last 15 years, the number of viruses and the different lesions associated with PVs in cats have greatly expanded. In this review, the PV life cycle and the subsequent immune response is briefly discussed along with methods used to investigate a PV etiology of a lesion. The seven PV types that are currently known to infect cats are reviewed. The lesions that have been associated with PV infections in cats are then discussed and the review finishes with a brief discussion on the use of vaccines to prevent PV-induced disease in domestic cats.