Common HPV Types Research Articles

High-risk human papillomavirus genotyping in cervical cancers in Tanzania

BackgroundHigh-risk human papillomavirus (hrHPV) infection causes almost all cervical cancer. Women living with human immunodeficiency virus (Women living with HIV: WLWHIV) are at a six-fold increased risk of developing cervical cancer. This study assessed hrHPV types in cervical cancer by HIV status and histologic subtypes at Muhimbili National Hospital (MNH) in Tanzania.MethodsThis cross-sectional study used formalin-fixed paraffin-embedded (FFPE) archived tissue blocks of cervical carcinomas diagnosed in the Department of Anatomical Pathology at MNH from January to December 2020. Tissue sections were tested for 15 HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, and 68) using the Ampfire assay. The distribution of HPV genotypes was assessed and compared by HIV status and histologic subtypes.ResultsThe mean age ± standard deviation (N = 227, with valid HPV results) was 55 ± 12.9 years, 28.6% (n = 65) were WLWHIV, and squamous cell carcinoma (SCC) was the most common histologic subtype (91.2%). Most cervical carcinomas (81.1%, n = 184) tested positive for hrHPV with HPV16 (44.1%), HPV18 (15.9%), HPV35 (8.4%) and HPV45 (5.7%) being the most common HPV types. hrHPV was higher among older women with 64.5%, 85.1% and 81.3% among 30–40, 41–60 and ≥ 61-year-old women, respectively (p = 0.033). HPV16 was more commonly detected in SCC (47.8%) than in adenocarcinomas (5%) (p < 0.0001). There was no difference in hrHPV positivity by HIV status.ConclusionsWe found a high proportion of hrHPV among cervical carcinomas diagnosed in Tanzania. Rolling out HPV vaccines that target more hrHPV types than HPV16/18, especially HPV35 and HPV45, could optimize protection against cervical cancer in Tanzania.

Type-specific human papillomavirus prevalence in women referred for colposcopy in Tehran.

Although several studies have been achieved on the frequency of the HPV types among women with cervical cancer in Iran, HPV-positive samples were in some cases directed to specific-primer genotyping of HPV 16 and 18. Therefore, the other HPV types are underestimated. Several studies have also reported a greater prevalence of HPV 16 in cervical cancer in Iran than in the world. To clarify these subjects, the distribution of HPV types in women referred for colposcopy in Tehran was investigated. In this cross-sectional study, a total of 148 cervical samples from women with normal, atypical squamous cells of undetermined significance, cervical intraepithelial neoplasia I-III, and invasive cervical cancer histopathology were included. HPV was detected by PCR assay and all HPV-positive specimens were subjected to direct nucleotide sequencing. Our results demonstrated that the total prevalence of HPV was 92.5%. The five most common HPV types were HPV 16 (49.3%), 18 (14.8%), 6 (7.4%), 31 (4.1%), and 11 (2.7%). About the histopathological stage, HPV 16 and 18 were dominant in all studied groups. In cervical cancer, HPV 16 and 18 were detected in 60% and 20% of cases, respectively. HPV 16 and 18 were the most common in cervical cancer in Iran.

Whole genome sequencing in high-grade cervical intraepithelial neoplasia patients from different ethnic groups in China.

Cervical cancer (CC) is the fourth most common cancer in women worldwide. It develops through precancerous lesions (cervical intraepithelial neoplasia (CIN), graded from low-grade (CIN1) to high-grade (CIN2-3)). It is well established that precancerous and cancerous cervical lesions are caused by a persistent infection with high-risk types of the human papilloma virus (hrHPV). To have a deeper understanding of the pathogenesis of CIN and CC, we systematically analyzed the landscape of genomic alterations and HPV integration profiles in high-grade CIN2/3. We performed deep whole genome sequencing on exfoliated cervical cells and matched peripheral blood samples from a cohort of 51 Chinese patients (of whom 35 were HPV+) with high-grade CIN from 3 ethnic groups and constructed strict integrated workflow of genomic analysis. In addition, the HPV types and integration breakpoints in the exfoliated cervical cells from these patients were examined. Genomic analysis identified 6 significantly mutated genes (SMGs), including CDKN2A, PIK3CB, FAM20A, RABEP1, TMPRSS2 and SS18L1, in 51 CIN2/3 samples. As none of them had previously been identified as SMGs in the Cancer Genome Atlas cervical squamous cell carcinoma and endocervical adenocarcinoma (TCGA-CESC) cohort, future studies with larger sample size of CINs may be needed to validate our findings. Mutational signature analysis showed that mutational signatures of CINs were dramatically different from CCs, highlighting their different mutational processes and etiologies. Moreover, non-silent somatic mutations were detected in all of the CIN2/3 samples, and 88% of these mutations occurred in genes that also mutated in CCs of TCGA cohort. CIN2 samples had significantly less non-silent mutations than CIN3 samples (P = .0006). Gene ontology and pathway level analysis revealed that functions of mutated genes were significantly associated with tumorigenesis, thus these genes may be involved in the development and progression of CC. HPV integration breakpoints occurred in 28.6% of the CIN2/3 samples with HPV infection. Integrations of common high risk HPV types in CCs, including HPV16, 52, 58 and 68, also occurred in the CIN samples. Our results lay the groundwork for a deeper understanding of the molecular mechanisms underlying the pathogenesis of CC and pave the way for new tools for screening, diagnosis and treatment of cervical precancerous and cancerous lesions.

Epidemiologic characteristics of high-risk HPV and the correlation between multiple infections and cervical lesions

BackgroundThe aim of this study was to determine the prevalence of high-risk human papillomavirus (HR-HPV) and the correlation between multiple infections and cervical lesions.MethodsThe current study involved population-based sample of 20,059 women who underwent cervical screening for 15 HR-HPV genotypes with ThinPrep cytologic test (TCT) results. The correlation between multiple HPV genotype infections and cervical lesions was also determined. The odds ratios (ORs) were calculated to assess co-infection patterns for each genotype with 15 other genotypes and the additive statistical interactions were evaluated.ResultsThere was a bimodal pattern among multiple HPV infections, with a peak in the younger group and a second peak in the elderly group. Indeed, most multiple HPV genotypes exhibited a bimodal pattern. The most common HPV type in patients with high-grade squamous intraepithelial lesions (HSILs) was HPV-16, followed by HPV-52, HPV-58, and HPV-33. The most frequent HPV type in patients with cervical cancer was HPV-16, followed by HPV-58 and HPV-33. Women with multiple infections were at a increased risk of low-grade squamous intraepithelial lesions [LSIL] (OR = 2.01; 95% CI 1.38–2.93) and HSIL (OR 2.28; 95% CI 1.36–3.81) when compared to women with single infections. patients with cervical cancer had the higher percentage of multiple HPV infections. Based on the data herein, we suggest that HPV-33 and HPV-58 may also be high-risk HPV types worthy of increased surveillance and follow-up. Conclusion: Our findings suggested that the association between multiple HPV infections and HSIL and LSIL are stronger compared to single HPV infections. There may be some specific combinations that synergistically affected the risk of HSIL and LSIL.

Human papillomavirus prevalence and type distribution in Chinese juvenile-onset recurrent respiratory papillomatosis patients

Objective: To evaluate HPV prevalence and type distribution in Chinese juvenile-onset recurrent respiratory papillomatosis (JoRRP) patients. Methods: We searched China National Knowledge Infrastructure, Wanfang data, China Biology Medicine disc, PubMed, Embase, and the Cochrane Library for studies assessing HPV infection of Chinese JoRRP patients up to 1 October, 2022. Two authors independently performed literature selection, data extraction, and quality assessment. HPV prevalence and HPV type-specific prevalence were pooled using a random effects model after Freeman-Tukey double arcsine transformation. All analyses were performed with R 4.1.3 software. Results: Nineteen publications investigating HPV infection of JoRRP patients were included in the final analyses. Of these, 16 studies reported HPV prevalence with a sample size of 1 528 patients, and 11 studies reported HPV6 prevalence and HPV11 prevalence with a sample size of 611 patients. All studies were graded as medium quality. In Chinese JoRRP patients, the synthesized HPV prevalence was 92.0% (95%CI:86.0%-96.6%, I2=87%), HPV6 prevalence was 42.4% (95%CI:34.9%-50.1%, I2=61%), and HPV11 prevalence was 72.3% (95%CI:59.0%-83.9%, I2=87%). All the pooled prevalence persisted in subgroup analyses stratified by publication year, sample size, and specimen type (P>0.05). There was no evidence of publication bias. In Chinese JoRRP patients, HPV16, 18, 31, 33, 52, and 58 prevalence was very low. Conclusions: Our findings suggested high HPV prevalence in Chinese JoRRP patients, and the most common HPV types were HPV6 and HPV11.

HPV integration as a prognostic biomarker for metastatic anal cancer: A next-generation sequencing ctDNA-based approach.

3525 Background: Anal cancer is an HPV-associated malignancy with increasing prevalence in the United States. Immune checkpoint blockade demonstrates limited efficacy in patients with metastatic squamous cell cancer of the anal canal (mSCCA), and no prognostic or predictive biomarkers have demonstrated clinical utility in this population. Available circulating tumor DNA (ctDNA) assays have utilized ddPCR methodologies to report only the most common HPV types (HPV-16 and HPV-18) and HPV copy numbers. To extend the utility of an HPV ctDNA assay, we created a novel next-generation sequencing HPV ctDNA assay to analyze HPV integration as a prognostic biomarker for patients with mSCCA. Methods: Using an IRB-approved protocol, ctDNA isolated from the plasma of patients with mSCCA was sequenced using a 1.4Mb hybrid-capture target-enrichment panel covering the whole genome sequences of all 193 HPV types. HPV type, HPV copy number, and HPV integration status/locus were determined using a bioinformatic pipeline developed in-house. A Fisher's exact test was used to compare radiographic and ctDNA changes in response to systemic treatment. Unpaired t-tests were used to compare demographics between patients with HPV integration (I) or no integration (NI) status in the ctDNA. Median survival was estimated (Kaplan Meier) and compared according to integration status using a log-rank test. Results: 68 plasma samples from 27 patients with mSCCA were analyzed. While HPV-16 was detected in ctDNA of all 27 patients, five patients had additional oncogenic HPV types [HPV-18 (2), HPV-45 (2), HPV-73 (1)]. Radiographic changes in metastatic disease volume were concordant with HPV copy number changes in the ctDNA (odds ratio 12.2, 95% confidence interval (CI) 2.0-75; p = .007). HPV integration events were detected in the ctDNA for 16 (59%) patients (median 2, range 1-7 unique events per patient). For the 23 patients with mSCCA who received anti-PD1 immunotherapy (15 I, 8 NI), there were no differences in age (60 vs 59 years, p = .88) or prior lines of treatment (3.0 vs 3.5, p = .72). There was a trend towards improved median progression-free survival for NI ctDNA status (5.8 vs 3.2 months, hazard ratio (HR) 1.8, 95% CI .71-4.5). Relative to NI status, I status was associated with worse overall survival rates (OS) (22.5 vs 44.9 months, HR 5.0, 95% CI 1.5-17, p = .005). Conclusions: Detection of HPV integration status for all HPV types is feasible using a ctDNA-based approach for patients with SCCA. HPV integration was prognostic for worse OS in patients with mSCCA. Identification of HPV integration as a novel prognostic biomarker for survival outcomes in mSCCA warrants further evaluation in larger clinical trials.

Determination of Human Papilloma Virus (HPV) Genotype Prevalance and Distrubution in Adana: A Hospital-Based Study Between 2014-2021

Cervical cancer is the fourth most common cancer among women all over the world. It is accepted that cervical cancer is highly related to the HPV. The International Agency for Research on Cancer (IARC) has classified 13 HPV types as group 1 carcinogens (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 66), which are commonly referred to as high risk-HPVs (hr-HPVs). Among these, hr-HPV-16 is undoubtedly the most carcinogenic based in the burden of cervical cancer (CC) and its precursor lesions. In our study, we analyzed retrospectively the data of a total of 2329 female patients who applied to the obstetrics and gynecology outpatient clinic of our hospital over a seven-year-period, whose cervical smear were carried out by the polymerase chain reaction (PCR) and cytology. In this study, it was aimed to determine the data of of HPV prevalence in our region during the seven-year-period from April 2014 to April 2021 and the most common genotypes and to interpret them together with the cervical smears cytology and biopsy results if it is available. HPV 3, 6, 11, 16, 18, 21, 26, 31, 33, 35, 39, 40, 42, 45, 51, 52, 53, 54, 55, 56, 58, 59, 61, 62, 66, 67, 68, 70, 72, 73, 81, 82, 83, 84 were identified by using linear array HPV genotyping test (Roche Diagnostics, Switzerland) from April 2014 to October 2017. HPV genotypes were identified by using HPV Genotypes 14 Real-TM Quant (Qiagen, Germany) between October 2017 and April 2021. This method detected HPV genotypes 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68. The data were analyzed using IBM SPSS Statistics (Version 25.0) predictive analytics software. Continuous variables are indicated as mean ± standard deviation, and discrete variables are indicated as number [percentage (%)]. Chi-square test is used to investigate dependencies between variables. All analyzes were evaluated to provide 95% confidence level and 80% test power. p<0.05 was accepted as significant for the analysis results. Out of 2329 patients, 1283 were found to be HPV negative (54.6%) and the others were found to be HPV positive (45.4%) by using real-time PCR in the cervical smears. It was detected that out of 1046 HPV positive patients, 585 of them (55.9%) had one HPV genotype and 461 of them (44.1%) had more than one HPV genotypes. As we divided all of the patients into two groups as <30 (Group I) ve > 30 (Group II) according to age range, HPV positivity was found 134/296 (45.2%) in Group I and 912/2033 (44.8%) in Group II. When we compared the HPV positive/negative results of Groups I and II by using chi-square test, no significant difference was found between the two age groups in terms of HPV positivity (p= 0.894). In our study, the most common HPV types were HPV 16 (14.2%), HPV 68 (8.2%), HPV 56 (8.2%), HPV 52 (7.1%), HPV 51 (6.8%), HPV 31 (6.5%), HPV 66(6.1%), HPV 39 (5.8%) and HPV 18 (5.6%) among the women with normal and abnormal cytology in the cervical smears. ASC-US was the most common abnormal epithelial cell change detected with HPV16 and 18 genotypes and it was detected 26.07% and 21.88% in patients, respectively. In our study, we found HPV prevalance in our region as 45.4% and the most common type was HPV 16. As a result, we concluded that it is important to determine regional HPV prevalance data, which is an important step in cervical cancer prevention strategies, and regional data of detected HPV genotypes.

Clinical and Genomic Determinants of Survival in Ugandan Women With Cervical Cancer

Clinical and Genomic Determinants of Survival in Ugandan Women With Cervical Cancer

Genotype distribution of cervical HPV among Caribbean women in a population-based study in Martinique: The DEPIPAPUFR study.

The Caribbean ranks seventh among the world regions most affected by cervical cancer. HPV-prevalence and genotype distributions also differ from regions. Knowledge of HPV genotype profiles is important for patients care and HPV vaccination implementation. The objective of this study was to describe HPV genotype distribution and risk factors in a population-based cohort of women in Martinique. In this study, 1312 women were included and underwent cervical cancer screening with successful sample collection between 2009 and 2014. Sociodemographic and clinical variables were recorded. Cytological examination of cervical vaginal smear was performed and classified(Bethesda). Detection of HPV DNA was performed with the PapilloCheck© Kit from Greiner Bio-one. Genotypes were analyzed for18 high-risk HPV (hrHPV) and 6low-risk HPV(lrHPV) types. A total of 1075 women were included with a mean age of 49.1±10.5 years. HPV prevalence was 27.6% (297/1075) with 19.4% (209/1075) women with only hrHPV, 5.3% (57/1075) with only lrHPV. Multiple infections (hrHPV/lrHPV) were detected in 31/240 cases of hrHPV (12.9%). A total of 353 hrHPV genotypes were analyzed; the most common HPV types were HPV51 (11.0%), HPV68 (10.8%), HPV53 (9.1%) and HPV 52 (7.1%). HPV16 and HPV18 represented respectively 4.8% and 4.0% of hrHPV genotypes. Abnormal cytology was observed in 34 cases (3.2%), with 14 ASCUS (1.3%), 10 LSIL (0.9%), 5 HSIL (0.5%), 3 ASC-H (0.3%) and 2 AGC (0.2%). Fifteen (44.1%) were hrHPV and 4 (14.7%) lrHPV; 7 cases of hrPHV were in the age-group 25–34 years. Among 1041cases of normal cytology, 225 had positive hrHPV detection (21.6%). This is the first population-based study of HPV profiles in our country, and we found a high prevalence of hrHPV. The most common genotypes were HPV51, 68, 53. These results could serve for cancer vaccination strategies and HPV surveillance in Martinique.

Prevalence and variety of HPV types in dependence on gender and age.

e17575 Background: Analysis of the typical structure of age, sex and territorial patterns of HPV spread is of interest, since it allows assessing the potential impact of modern immunoprophylaxis. The purpose of the study was to analyze the prevalence and variety of HPV types in dependence on gender and age. Methods: The study included 424 patients (334 women and 90 men) under 45 years of age (228 women and 46 men) and over 46 years old (106 and 44, respectively). Vaginal and cervical swabs were examined in women and urethral swabs in men. DNA extraction was performed by Real-time PCR. Results: HPV DNAs were found in 150(35.4%) patients, including 115(34.4%) women and 35(38.9%) men (p &gt; 0.05). HPV was less frequently detected in older women, and more frequently in older men. Thus, HPV DNAs were found in 85(37.3%) women &lt; 45 years and in 30(28.3%, p = 0.068) women &gt;46 years; in men – 14(30.9%) and 21(47.7%, p = 0.071), respectively. Some HPV types were more frequent in younger patients, compared to older ones: in 26(26.3%) and in 5(9.8%), p = 0.068) of HPV-positive patients, respectively. The most common HPV types were 16(46.7%), 52(16.0%), 56(12.7%), 31(10.7%); HPV16 dominated both in females - 50(50.4%) and males - 12(34.6%). After HPV16, most frequent types in women were: 31-12(11.3%) and 52-13(11.3%), 18-12(10.4%) and 56-119(, 6%); in men - 52-11(31.4%), 56-8(22.9%), 45-4(11.4%) and 18-3(8.6%) types. The total share of two HPV types with the greatest oncogenic potential (types 16 and 18) was 85(56.7%) of all HPV-positive patients, and in women it was higher than in men: 70(60.9) vs. 15(42.9%, p = 0.046), respectively. Conclusions: Simultaneous infection with several HPV types was statistically significantly more often detected in younger patients compared with the older age group among men. HPV16 was the most common type both in men and women; however, the rates of other HPV types differed. Cumulative frequency of types 16 and 18 was statistically significantly higher in women, compared to men.

HPV58 E7 Protein Expression Profile in Cervical Cancer and CIN with Immunohistochemistry.

Background: The persistent infection of high-risk human papillomavirus (HR-HPV) is one of the most common causes of cervical cancer worldwide, and HPV type 58 (HPV58) is the third most common HPV type in eastern Asia. The E7 oncoprotein is constitutively expressed in HPV58-associated cervical cancer cells and plays a key role during tumorigenesis. This study aimed to assess the HPV58 E7 protein expression in the tissues of cervical cancer and cervical intraepithelial neoplasia (CIN).Methods: A total of 67 HPV58-positive cervical samples were collected, including 25 cervical cancer samples and 42 CIN samples. All the tissues were examined by HPV58 E7, p16INK4a and Ki67 immunohistochemistry (IHC). At last, we analyzed their association with clinical and pathological variables.Results: HPV58 E7 expression was detected in 96% of the HPV58 DNA-positive cervical cancer tissues and 85.7% of HPV58-positive CIN tissues. 65 samples of cervical cancer and CIN tissues had p16-positive staining, while 59 samples were Ki-67 positive.Conclusions: HPV58 E7 protein is highly expressed in both cervical cancer and CIN tissues. HPV58 E7 IHC could be sensitive and specific for evaluating HPV-driven cervical cancer and pre-cancerous lesions, in combination with p16 and Ki-67 IHC.

Abstract PO-087: Anal cancer and its association with HPV16 lineage B in African Americans

Abstract Background: Human papillomavirus type 16 (HPV16) is one of the most common and carcinogenic HPV types associated with high risk of anal, vagina, vulva, penis and cervical neoplastic transformations. However, many genetic variants exist within this virus and not all seem to have the same carcinogenic potential. Aim: To determine HPV16 lineages and their association with risk of high-grade anal lesions in African Americans in an inner-city hospital. Methods: We reviewed medical records of 370 African Americans with anal lesions from Jan. 2007 to Dec. 2015. This study was approved by Howard University Institutional Review Board. Demographic, clinical and pathological data including HPV, HIV, HCV (hepatitis C virus), diabetes mellitus, hypertension and body mass index (BMI) were collected. DNA was extracted from a subset of HPV16-positive patients with FFPE tissue samples (72 patients, 111 samples) and used for HPV16 whole-genome sequencing. We assessed HPV16 variant lineages and associations with disease stage. Statistical analyses were performed using Chi-square tests, Student’s t-tests, and logistic regression. Odds ratios (OR) and p-values were calculated for comparisons of normal/condyloma/high-grade dysplasia (HGD) vs. squamous cell carcinoma (SCC) and for normal/condyloma vs. HGD. The most common HPV16 A1 sublineage was used as a reference in these comparisons. Results: Males represented 75% of the patients (n=276), with a median age of 44 years and BMI of 25.8 kg/m2. The frequency of condyloma, high-grade dysplasia, SCC and adenocarcinoma was 191 (52%), 26 (7%), 31 (8%) and 8 (2%), respectively. The frequency of HPV, HIV, and HCV was 231 (68%), 147 (43%) and 42 (12%), respectively. HPV and HIV were risk factors for condyloma and dysplasia (P&amp;lt;0.05). All four main lineages of HPV16 (A,B,C,D) were detected in our specimens, with sublineage A1 most common. Lineage B, also named the African-1 HPV16 lineage because it is most common in Africa, had the strongest association with SCC (OR=10.5) whether alone or in combination with lineages A4 and D (OR=10.5), although with a lower statistical significance (0.054 vs. 0.009). As for HGD, lineage B along with A4, C and D only gave an OR of 1.4. Conclusion: We show that the majority of patients with anal lesions are young males with HPV and HIV co-infections. HPV16 lineage B was associated with a high risk of SCC development. Citation Format: Hassan Ashktorab, Adeyinka Laiyemo, Lisa Mirabello, Hassan Brim. Anal cancer and its association with HPV16 lineage B in African Americans [abstract]. In: Proceedings of the AACR Virtual Conference: Thirteenth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2020 Oct 2-4. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(12 Suppl):Abstract nr PO-087.

Novel concepts in cervical cancer screening: a comparison of VIA, HPV DNA test and p16INK4a/Ki-67 dual stain cytology in Western Kenya

BackgroundScreening of unvaccinated women remains essential to mitigate the high morbidity/mortality of cervical cancer. Here, we compared visual inspection with acetic acid (VIA), recommended by WHO as the most cost-effective screening approach in LMICs, with HPV-based screening, and usage of p16INK4a/Ki-67 dual stain cytology.MethodsWe prospectively enrolled women participating in a VIA-based cervical cancer screening program in two peri-urban health centers of Kenya. Consenting women had a VIA examination preceded by collection of a liquid-based cytology sample from the cervix stored in PreservCyt medium (Hologic®). Analysis of all samples included a hrHPV DNA test and evaluation of a p16INK4a /Ki-67 (CINtecPLUS®) dual stained slide that was prepared using the ThinPrep® 2000 Processor and evaluated by a pathologist trained in the methodology.ResultsIn 701 of a total of 800 women aged 18–64 years, all three investigations were performed and data could be analyzed. The HPV, VIA and dual stain cytology positivity were 33%, 7%, and 2% respectively. The HPV positivity rate of VIA positive cases was 32%. The five most common HPV types were HPV16, 52, 68, 58 and 35. The OR among HIV infected women of an HPV infection, VIA positivity and positive dual stain cytology were 2.6 (95%CI 1.5–4.3), 1.9 (95%CI 0.89–4.4) and 3.4 (95%CI 1.07–10.9) respectively. The sensitivity of VIA to detect a p16INK4a/Ki-67 positive transforming infection was 13% (95%CI 2–38).ConclusionsPrimary HPV testing appears feasible and should be considered as a primary screening test also in LMICs. The poor sensitivity of VIA renders it unsuitable as a triage test for HPV positive women. The utility of p16INK4a/Ki-67 dual stain cytology as a triage test for HPV positive women in LMICs should be further studied.

Abstract 2027: Anal cancer among African Americans associate with HPV16 lineage B and HIV

Abstract Background: Human papillomavirus type 16 (HPV16) is one of the most common and carcinogenic HPV types associated with high risk of anal, vagina, vulva, penis and cervical neoplastic transformations. However, many genetic variants exist within this virus and not all seem to have the same carcinogenic potential. Aim: To determine HPV16 lineages and their association with risk of high-grade anal lesions in African Americans in an inner-city hospital. Methods: We reviewed medical records of 370 African Americans with anal lesions from Jan. 2007 to Dec. 2015. This study was approved by Howard University Institutional Review Board. Demographic, clinical and pathological data including HPV, HIV, HCV (hepatitis C virus), diabetes mellitus, hypertension and body mass index (BMI) were collected. DNA was extracted from a subset of HPV16-positive patients with FFPE tissue samples (72 patients, 111 samples) and used for HPV16 whole-genome sequencing. We assessed HPV16 variant lineages and associations with disease stage. Statistical analyses were performed using Chi-square tests, Student's t-tests, and logistic regression. Odds ratios (OR) and p-values were calculated for comparisons of normal/condyloma/high-grade dysplasia (HGD) vs. squamous cell carcinoma (SCC) and for normal/condyloma vs. HGD. The most common HPV16 A1 sublineage was used as a reference in these comparisons. Results: Males represented 75% of the patients (n=276), with a median age of 44 years and BMI of 25.8 kg/m2. The frequency of condyloma, high-grade dysplasia, SCC and adenocarcinoma was 191 (52%), 26 (7%), 31 (8%) and 8 (2%), respectively. The frequency of HPV, HIV, and HCV was 231 (68%), 147 (43%) and 42 (12%), respectively. HPV and HIV were risk factors for condyloma and dysplasia (P&amp;lt;0.05). All four main lineages of HPV16 (A,B,C,D) were detected in our specimens, with sublineage A1 most common. Lineage B, also named the African-1 HPV16 lineage because it is most common in Africa, had the strongest association with SCC (OR=10.5) whether alone or in combination with lineages A4 and D (OR=10.5), although with a lower statistical significance (0.054 vs. 0.009). As for HGD, lineage B along with A4, C and D only gave an OR of 1.4. Conclusion: We show that the majority of patients with anal lesions are young males with HPV and HIV co-infections. HPV16 lineage B was associated with a high risk of SCC development. Citation Format: Hassan Brim, Lisa Mirabello, Ali Afsari, Muneer Abbas, Meredith Yeager, Joseph Boland, Sara Bass, Mia Steinberg, Michael Cullen, Adeyinka Laiyemo, Tammy Naab, Babak Shokrani, Edward Lee, Mehdi Nouraie, Hassan Ashktorab. Anal cancer among African Americans associate with HPV16 lineage B and HIV [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2027.

How to triage HPV positive cases: Results of four million females

ObjectiveTo evaluate the Turkey's nationwide HPV DNA screening program on the basis of first 4 million screened women. MethodsWomen over age 30 were invited for screening via HPV DNA and conventional cytology. Single visit screen strategy was used to collect for both screening and triage (extended genotyping and conventional pap-smear). ResultsA total of 4,099,230 patients had attended to HPV DNA cancer screening. 4.39% were found to be HPV DNA positive. The most common HPV type was 16, followed by 51, 31, 52, 56 and 18 at all age intervals and geographic regions. Cytology results were reported as “normal” in (69.2%), “inadequate sampling” in (16.6%) and as “abnormal (≥ASC-US)” in the remaining. Current Turkish screening with HPV DNA (referral to colposcopy with HPV 16 or 18 or any smear abnormality ≥ASC-US) gives overall PPV of 24.3% for ≥CIN2. Only Pap-Smear triage revealed PPV of 26.4% for ≥ASC-US thresholds. Comparison of different triage methods for ≥CIN2+ according to different HPV genotype revealed a PPV of 32,6% for HPV 16; 15,3% for HPV 18. This figure was 34.4%, 19.3%, 15.3% and 14.0% for HPV 33, 31, 45 and 35; respectively. ConclusionThis study involves the largest series in the world summarizing a real-world experience with primary HPV DNA screening and triage with a single visit. The results show the feasibility and applicability of such screening method in developing countries with acceptable colposcopy referral rates. Among triage tests, only pap-smear seems to be effective without a need for extended genotyping.

Abstract A17: Human papillomavirus type 16 and 18 viral loads as predictors associated with abnormal cervical cytology among women in Saudi Arabia

Abstract The present study aimed to screen for and genotype HPV among Saudi women; develop sensitive quantitative polymerase chain reaction (qPCR) assays to detect the viral load for the two most common HPV types, namely 16 and 18; and assess whether HPV viral load could be used as a marker for cervical abnormality and disease progression. This study examined 733 specimens (both formalin-fixed, paraffin-embedded specimens and PAP smear samples) from women who underwent cervical screening. The specimens and samples were processed for DNA extraction and then tested for HPV DNA. Approximately 165 specimens (18%) were positive for HPV. Those specimens were genotyped using a reverse line blotting hybridization assay. The results indicated that the most common HPV types detected were a single infection with HPV 16 (51%) or with HPV 18 (28%) followed by infections with multiple HPV types (~7%). A qPCR TaqMan assay developed in-house was used to determine the viral load for HPV genotypes 16 (n=80) and 18 (n=45). Viral loads for both HPV types were significantly associated with cervical cytology grade (P &amp;lt; 0.05). The odds ratio (OR) for the HPV 16 viral load was high for specimens with cervical cancer (OR, 18.8; 95% CI, 4.3–82.9) or for those with high-grade squamous intraepithelial lesions (OR, 14.7; 95% Cl, 2.43–88.49). For the HPV 18 viral load, the OR was significant only for specimens with cervical cancer (OR, 11.1; 95% Cl, 2.2–54.9). Logistic regression models for HPV 16 and for HPV 18 viral load levels were significant, with higher viral load associated with cervical abnormalities. These findings indicate that viral load is a predictor significantly associated with cytology abnormality in women who are positive for high-risk HPVs and suggest integrating a viral load test into current clinical screening practices for HPV-positive women. Citation Format: Dalia Obeid, Shihana Almatrrouk, Hadeel Khayat, Turfah Al-Muammer, Asma Tullbah, Ismail Albadawi, Mohammed Alahdal, Fatimah Alhamlan. Human papillomavirus type 16 and 18 viral loads as predictors associated with abnormal cervical cytology among women in Saudi Arabia [abstract]. In: Proceedings of the AACR Special Conference on the Microbiome, Viruses, and Cancer; 2020 Feb 21-24; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2020;80(8 Suppl):Abstract nr A17.

Clinical and epidemiological features of papillomavirus infection manifestations on the example of cervical cancer and genital warts

Background: Cervical cancer and genital warts (GWs) are some of the most common manifestations of human papillomavirus infection (HPV). These lesions cause significant damage to the reproductive health of the population, which leads to increased attention to the prevention of HPV infection among various population groups.&#x0D; Aims: To determine the clinical and epidemiological features of the HPV manifestations by the example of cervical cancer and genital warts.&#x0D; Methods: A retrospective analysis of anamnestic information of 115 women with an established diagnosis of cervical cancer and 177 patients with an established diagnosis of GWs was performed. The clinical and epidemiological characteristics of patients with diagnoses of GWs and cervical cancer were based on the development of outpatient admission cards and inpatient histories, as well as test data for HPV.&#x0D; Results: HPV 16 was the most common HPV type among patients with GWs and cervical cancer ― it was detected in 37.6% of cases. Also the most frequently encountered: HPV 6/18/11/31/51/52. In 43.2% cases of HPV detection, two or more types were detected at once, the most common combinations: HPV16 and HPV18, HPV6 and HPV16, HPV6 and HPV11. Analysis of the frequency of screening for cervical cancer and visits to the gynecologist for 5 years before establishing the diagnosis showed that among those who did not screen for cervical cancer, the risk of diagnosing stage IIIV was 5.2 times higher than among individuals who underwent cervical screening 2 years ago, or once a year for the last five years. Among patients with GWs who had 2 or more sexual partners for 1 year, 13.5% of patients regularly used barrier contraception methods (condoms) during sexual contact, not regularly ― 61.5%, did not use them at all ― 25.0%.&#x0D; Conclusions: Identifying the clinical and epidemiological features of HPV infection should contribute to the development of new and optimize existing prevention programs for a wide range of HPV-associated diseases.

Human papillomavirus type 16 and 18 viral loads as predictors associated with abnormal cervical cytology among women in Saudi Arabia

The detection of HPV viral DNA is regularly conducted with cervical screening. However, using a molecular marker such as the viral load may serve as a predictor associated with disease detection and progression. The present study aimed to screen for and genotype HPV among women in Saudi Arabia, develop and validate sensitive quantitative polymerase chain reaction (qPCR) assays to detect viral load for the two most common HPV types, namely 16 and 18, and assess whether HPV viral load could be used as a marker for cervical abnormality and disease progression. This study examined 733 specimens (both formalin-fixed paraffin embedded specimens and PAP smear samples) from women who underwent cervical screening. The specimens and samples were processed for DNA extraction and then tested for HPV DNA using nested PCR. Approximately 165 specimens (18%) were positive for HPV. Those specimens were genotyped using a reverse line blotting hybridization assay. The results indicated that the most common HPV types detected were a single infection with HPV 16 (51%) or with HPV 18 (28%) followed by infections with multiple HPV types (~7%). A qPCR TaqMan assay developed and validated in-house was used to determine viral load for HPV genotypes 16 (n = 80) and 18 (n = 45). Viral loads for both HPV types were significantly associated with cervical cytology grade (P < 0.05). The odds ratio (OR) for the HPV 16 viral load was high for specimens with cervical cancer (OR, 18.8; 95% CI, 4.3–82.9) or for those with high-grade squamous intraepithelial lesions (OR, 14.7; 95% Cl, 2.43–88.49). For the HPV 18 viral load, the OR was significant only for specimens with cervical cancer (OR, 11.1; 95% Cl, 2.2–54.9). Logistic regression models for HPV 16 and for HPV 18 viral load levels were significant, with higher viral load associated with cervical abnormalities. These findings indicate that viral load is a predictor significantly associated with cytology abnormality in women who are positive for high-risk HPVs and suggest that integrating a viral load test into current clinical screening practices for HPV-positive women is warranted in Saudi Arabia.

Progression Low Squamous Intraepithelial Lesion and Human Papillomavirus Infections.

Introduction:Infection with human papillomavirus is the main cause of cervical carcinoma. In Bosnia and Herzegovina (BIH) 556 cases of cervical carcinoma are diagnosed annually, and 141 women die from it.Aim:To determine the most common HPV type found in ASCUS and LSIL and progression, regression and persistence of lesions.Methods:In a retrospective study, 11 051 PAP tests, performed in several private gynecological practices located in Tuzla Canton from January 2016 to December 2019, were analyzed. In processing of data , X2 – statistical method was used.Results:11051 PAP test were inspected. Normal findings were found in 90.48% (N-10002) and pathological findings in 9.49% (N- 049). ASCUS was present in 4.9% (N-544), LSIL in 3.04% (N-337), HSIL in 0.74% (N-84), ASC-H in 0.27% (N-30) and AGC in 0.49% (N-55) of cases. The most common is HPV 16, found in 50.5% (N-44) of ASCUS and LSIL. Monoinfection with HPV 16 was found in 40.9% (N-18) ASCUS (N-3) and LSIL (N-15), and a combination of HPV 16 with other types like HPV 18, 31, 33, 39 in 59% (N-26). Progression of ASCUS lesion in HSIL 1.6% (N-2), and LSIL 9.6% (N-12). Progression of LSIL to HSIL was found in 9.0% (N-10). HSIL progresses significantly more frequent from LSIL (p<0.05) than from ASCUS changes. Lesions that progress into higher grade HSIL are HPV 16 positive. Progression into HSIL is not found in patients with low-risk HPV 6 and 11 infection.Conclusion:Women infected with HPV 16 have more a frequent progression of a lesion into higher grade HSIL. They should be intensively monitored because of the increased risk for development of cervical carcinoma.

Distribution of high-risk α-genus human papillomavirus genotypes impacts cutaneous neoplasms.

High-risk α-genus human papillomaviruses (α-HPVs) are linked to cervical and genital carcinomas; however, their correlation with cutaneous squamous cell carcinoma (cuSCC) or premalignant skin lesions remains controversial. We evaluated the contribution of high-risk α-HPV to the occurrence of cuSCC, Bowen's disease and actinic keratosis (AK), and the distribution of high-risk α-HPV genotypes in these cutaneous tumours. HPV genotypes were determined using a commercial PCR-based microarray on skin tissue samples collected from 76 [38 young (<60years) and 38 elderly (>60years)] cuSCC, 34 Bowen's disease, 48 AK patients and 10 young controls. Associations between α-HPV prevalence and relevant risk factors were analysed. High-risk α-HPV was more frequently detected in cuSCC patients (57.9%) than in the patients with Bowen's disease (38.2%), AK (0.0%) and control patients (10.0%). The high-risk α-HPV prevalence was higher in young than in elderly cuSCC patients (65.8% vs. 50.0%, P=0.031). The most common HPV type was 16, present in 90.9% of all HPV-carrying cuSCC patients. Multiple infections with different high-risk α-HPV types were found in 20.5% of HPV-related cuSCC, whereas only single infection with type 16 was found in Bowen's disease. Although sun exposure is known as a major risk factor for cuSCC, high-risk α-HPVs were more frequently found in non-exposed sites rather than in sun-exposed sites of cuSCC. Multiple infections, as well as single infection with high-risk α-HPV may link to cuSCC. In spite of the involvement of high-risk α-HPV at high levels in cuSCC and Bowen's disease, no high-risk α-HPV was detected in AK patients, suggesting that Bowen's disease rather than AK might be involved in the development of HPV-related cuSCC as a precursor.