Oral squamous cell carcinoma (OSCC) often presents as an invasive tumor with poor prognosis. Recent evidence suggests that microRNAs, particularly miR-1-3p, may regulate the molecular pathways associated with tumor invasion. This study aimed to investigate the functional role of miR-1-3p in OSCC invasion using integrated in vitro, in vivo, and ex vivo approaches, and to evaluate its expression in tumors with varying depths of invasion (DOI). SCC-9 cells were transfected with miR-1-3p mimic and inhibitor. Invasion was assessed using transwell assays. Zebrafish xenografts were generated to evaluate tumor area, invasion depth, migration, and metastasis. Additionally, miR-1-3p expression was quantified by RT-qPCR in 26 human OSCC samples stratified by different groups of DOI. miR-1-3p inhibition significantly increased tumor cell invasion in vitro (P = 0.039) and resulted in larger, deeper, and more migratory tumors in vivo (P < 0.05). All in vitro experiments were performed in duplicate, which may limit the precision of quantitative estimates. Metastases were observed in 30.8

High-risk human papillomaviruses (HPV) are the main etiological agents of cervical cancer. The viral early promoter regulates the expression of E6/E7 oncoproteins, and its transcriptional activity is positively or negatively regulated by host transcription factors (TFs) that are able to bind to the viral long control region (LCR). In this study, we assessed the impact of a TF library on the early transcriptional activity of high-risk HPV-16 and -18, and among these TF, we selected and investigated the impact of GATA3, as well as its potential role as a prognostic biomarker for the development of cervical cancer. Luciferase reporter assays demonstrated that GATA3 negatively influences the transcriptional activity of HPV-16 and -18, downregulating E6 and E7 mRNA levels, with in silico and in vivo assays indicating that this effect is due to GATA3 direct binding to the viral LCR. Subsequently, we evaluated GATA3 levels in normal and HPV-immortalized epithelial raft cultures and cervical cancer samples by immunohistochemistry, also accessing the TF presence in pre-neoplastic intraepithelial cervical lesions (CIN) by immunofluorescence assays, further correlating GATA3 protein expression with the presence of HPV E6/E7 mRNA. This approach revealed an apparent inverse correlation between GATA3 expression and the grade of CIN lesions, as well as with the presence of viral oncogene transcripts. When accessing GATA3 expression in cancer samples, we observed a significant correlation between the absence of GATA3 and higher stages of cancer. Thus, our data indicates that the loss of GATA3 expression contributes to high-risk HPV-mediated cervical carcinogenesis, with the TF possibility acting as a protective factor whose absence enables sustained viral oncogene expression and disease progression in the cervical tissue.

IntroductionSolid oral tumors represent approximately 6% of malignant neoplasms in dogs, with melanoma, squamous cell carcinoma, and fibrosarcoma identified as the most prevalent types. This case-control study examined internal and external factors associated with their development.MethodsOwners of 80 dogs with oral tumors and 95 healthy controls completed a structured epidemiological questionnaire. Univariate and multivariate analyses were performed.ResultsSignificant associations were identified regarding age, weight, reproductive status, dietary habits, and environmental exposures, such as barbecue smoke and professional dental cleaning.DiscussionThese findings suggest potential risk factors, though the study’s observational design requires cautious interpretation. Further investigation through prospective studies is warranted.

Forkhead box protein A1 (FOXA1) is linked to cancer development in different anatomical sites. We previously demonstrated that FOXA1 markedly increases high-risk human papillomavirus (HPV) transcription by directly binding to the long control region (LCR). Herein, we aimed to assess FOXA1 levels and its prognostic significance in cervical cancer (CC). Methods: Patients with CC treated at the Instituto do Cancer do Estado de São Paulo (ICESP), Brazil, were included. FOXA1 levels were measured using immunohistochemistry (IHC). Results: Overall, FOXA1 staining was observed in 85% (208/245) of the samples, primarily localized in both the nucleus and cytoplasm (71%). High FOXA1 staining was more frequently detected in squamous cell carcinoma (SCC) compared to adenocarcinomas (ADC) (p = 0.003). High FOXA1 staining was significantly associated with HPV-positive CC (p = 0.005), with worse overall survival (OS) and disease-free survival (DFS) when considering all CC samples, as well as when restricting the analysis to SCC cases. Among HPV-positive cases, high FOXA1 staining was also significantly associated with poorer OS or DFS in women with SCC. Finally, high FOXA1 staining was associated with poor OS (HR 1.79; 95% CI: 1.14–2.79; p = 0.011) and poor DFS (HR: 1.64; 95% CI: 0.98–2.74; p = 0.056) among SCC cases in comparison to cases with low/moderate staining. No association was found between FOXA1 levels and prognosis in ADC. Conclusions: Our data suggest that FOXA1 may be involved in HPV-induced carcinogenesis and could serve as a prognostic biomarker in patients with cervical SCC.

BACKGROUND:Circulating tumor DNA (ctDNA) is a promising biomarker to predict recurrence in solid tumors such as colorectal and breast cancer. Defining complete response after chemoradiation remains challenging, as post-treatment inflammatory and tissue changes limit clinical and imaging evaluation. METHODS:The authors conducted a prospective longitudinal study of patients with localized cervical and anal cancer treated with curative-intent chemoradiation with the aim of evaluating the role of ctDNA as a predictor of recurrence. Blood specimens were collected at D1, D29, immediately post-treatment, 8-weeks, 24-weeks, and every 6-months up to 5-years. ctDNA testing (Signatera™) was provided by Natera as an in-kind donation. The primary endpoint was correlation between 8-week ctDNA and recurrence. RESULTS:28 patients were evaluable. Ten recurrences occurred, all in patients with positive ctDNA after chemoradiation and before radiologic progression. The median interval between ctDNA positivity and radiologic progression was 3.0-months (range 0.5-21.3). All patients with radiologic complete response at 6 months had negative 8-week ctDNA. Among those without complete response, 8-week ctDNA identified most who later progressed (6/8), while all without complete response who did not progress had negative ctDNA. Eight-week ctDNA positivity was strongly associated with inferior progression-free survival: median 6.7-months in ctDNA-positive versus not reached in ctDNA-negative patients (HR = 40.09, p = 0.001). CONCLUSION:Among patients with cervical and anal cancer, ctDNA monitoring after definitive chemoradiation identifies individuals without complete response and predicts recurrence. ctDNA positivity at 8-weeks identifies a very high-risk group for whom treatment intensification trials are warranted.

Cervical cancer remains a leading cause of mortality among women, particularly in low- and middle-income countries. Despite distinct prognoses and clinical outcomes between its main histological subtypes, squamous cell carcinoma (SCC) and adenocarcinoma (ADC), current treatment regimens remain largely similar, creating an urgent need for targeted therapeutic strategies based on molecular distinctions. To address this gap, the long noncoding RNA IGFL2-AS1 was investigated as a potential prognostic biomarker and therapeutic target in cervical adenocarcinoma. A translational approach was employed that integrated patient transcriptome data, in silico analysis from public databases, and in vitro validation. Using CRISPR/dCas9 technology, IGFL2-AS1 expression was modulated in HeLa (ADC) and SiHa (SCC) cell lines to assess its impact on cellular characteristics associated with tumorigenesis. In silico analysis revealed that IGFL2-AS1 expression was significantly reduced in ADC compared to SCC, and its low expression was consistently linked to poorer ADC prognosis and decreased patient survival. Notably, overexpression of IGFL2-AS1 in HeLa cells significantly reduced cell proliferation, migration, clonogenic survival, and enhanced sensitivity to cisplatin and doxorubicin. Conversely, IGFL2-AS1 repression in SiHa cells yielded no significant phenotypic changes, suggesting a context-dependent mechanism. IGFL2-AS1 is identified as a histological subtype-specific prognostic biomarker and promising therapeutic target for cervical adenocarcinoma.

BACKGROUND:Forkhead box protein A1 (FOXA1) is linked to cancer development in different anatomical sites. We previously demonstrated that FOXA1 markedly increases high-risk human papillomavirus (HPV) transcription by directly binding to the long control region (LCR). Herein, we aimed to assess FOXA1 expression and its prognostic significance in cervical cancer (CC). METHODS:Patients with CC treated at the Instituto do Cancer do Estado de São Paulo (ICESP), Brazil, were included. FOXA1 levels were measured using immunohistochemistry (IHC). RESULTS:Overall, FOXA1 staining was observed in 85% (208/245) of the samples, primarily localized in both the nucleus and cytoplasm (71%). High FOXA1 staining was more frequently detected in squamous cell carcinoma (SCC) compared to adenocarcinomas (ADC) (p = 0.003). High FOXA1 staining was significantly associated with HPV-positive CC (p = 0.005). It was also associated with worse overall survival (OS) and disease-free survival (DFS) when considering all CC samples, as well as when restricting the analysis to SCC cases. Among HPV-positive cases, high FOXA1 staining was also significantly associated with poorer OS and DFS in women with SCC. Finally, high FOXA1 staining was associated with poor OS (HR 1.79; 95% CI: 1.14-2.79; p = 0.011) and poor DFS (HR: 1.64; 95% CI: 0.98-2.74; p = 0.056) among SCC cases in comparison to cases with low/moderate staining. No association was found between FOXA1 levels and prognosis in ADC. CONCLUSIONS:Our data suggest that FOXA1 may be involved in HPV-induced carcinogenesis and could serve as a prognostic biomarker in patients with cervical SCC.

Objectives. To evaluate the prevalence and genotype distribution of human papillomavirus (HPV) and their association with cervical cytologic abnormalities among women undergoing routine cervical cancer screening in the Vale do Ribeira region, São Paulo, Brazil.Methods. Liquid-based cytology samples from 1,502 women attending primary healthcare units between September 2020 and December 2022 were analyzed. HPV detection and genotyping were performed using PGMY09/11 PCR followed by Sanger sequencing. Associations were evaluated using chi-square or Fisher’s exact tests and Poisson regression with robust variance.Results. HPV DNA was detected in 11.1% of women, and high-risk HPV (HR-HPV) was identified in 6.5%. HR-HPV prevalence increased significantly with cytologic severity, rising from 4.3% in women with negative cytology (NILM) to 25.5% in those with low-grade abnormalities (ASC-US/LSIL) and 40.0% in those with high-grade abnormalities (AGC/ASC-H/HSIL) (p < 0.001). HPV-16 (18.8%), HPV-52 (10.7%), HPV-51 (7.4%), HPV-58 (5.4%), and HPV-35 (5.4%) were the most frequent HR-HPV types. HPV-58 was the most common genotype among women with high-grade cytologic abnormalities. In multivariable analysis, women aged 25–44 years (PR=0.60; 95% CI:0.38–0.95) and ≥45 years (PR=0.40; 95% CI:0.24–0.68) had lower HPV prevalence than women aged 18–24 years, whereas reporting 3–4 (PR=1.81; 95% CI:1.21–2.70) or ≥5 lifetime sexual partners (PR=1.87; 95% CI:1.25–2.81) was independently associated with HPV positivity.Conclusions. HR-HPV prevalence increased markedly with cytologic severity. Although HPV-16 predominated overall, HPV-58 was more frequent in high-grade abnormalities, highlighting regional genotype patterns relevant to HPV-based screening and vaccination strategies.

Human papillomavirus Type 16 (HPV16) variants are classified into four lineages (A-D) and 16 sublineages: A1-3 (European), A4 (Asian), B1-4 and C1-4 (African), and D1-4 (Asian-American). This study investigated the genetic HPV16 variability through sequencing of the complete long control region (LCR), in 44 cervical samples from Paraguayan women, including 19 without cervical intraepithelial neoplasia grade 2 (<CIN2) and 25 with CIN2 or worse (CIN2+: 1 CIN2, 16 CIN3, and 8 invasive cancers). Sublineages A1 (81.8%), A4 (2.3%), D2 (2.3%), D3 (11.4%), and D4 (2.3%) were detected. A total of 48 point mutations were detected, of which 11 showed nominal association with CIN2+ (p < 0.05), although none remained statistically significant after correction for multiple comparisons. Notably, most of these mutations were predominantly observed in non-A1-A3 variants. An exploratory association was observed between non-A1-A3 variants and the presence of CIN2+ ( p = 0.0065). Functional analysis revealed that the A4 variant, detected in a woman with cancer, exhibited the highest transcriptional activity (2.34 ± 0.78), followed by D3 (0.72 ± 0.21) and A1 (0.03 ± 0.02), corresponding to samples from women with CIN3 and without lesions, respectively. In conclusion, we report for the first time the detection of an A4 HPV16 variant in Paraguay, which exhibited the highest transcriptional activity among the variants analyzed. These findings suggest potential functional differences among HPV16 variants and are consistent with a possible relevance of non-A1-A3 variants in cervical lesion severity. However, these results should be interpreted with caution as exploratory and hypothesis-generating.

TPS5621 Background: Cervical and anal cancers treated with definitive radiotherapy (RT), with or without chemotherapy, frequently exhibit persistent disease or recurrence. Post-treatment imaging—the current gold standard for assessing response—faces limitations due to local inflammatory processes, necessitating repeated imaging and invasive biopsies, which increase complexity and costs. Circulating tumor DNA (ctDNA) has emerged as a promising biomarker for assessing recurrence or disease progression in HPV-associated cancers. This study proposes the development of a low-cost HPV ctDNA test. Additionally, it aims to evaluate ctDNA’s accuracy compared to standard imaging and its role in guiding immunotherapy for patients at high risk of recurrence. Methods: ANA trial seeks to establish a novel, affordable diagnostic approach to optimize follow-up and treatment strategies for HPV-associated cancers, potentially improving outcomes while reducing costs. This is a prospective, single-center study with two components: 1. Non-interventional phase: Development and validation of a low-cost HPV ctDNA test. The test’s accuracy will be compared with commercially available ctDNA tests and standard imaging in monitoring patients with HPV-associated cervical and anal cancers post-definitive RT or chemoradiotherapy. 2. Interventional phase (Phase II trial): A single-arm study evaluating the efficacy of early complementary immunotherapy in patients with persistent ctDNA positivity post-treatment. Eligibility criteria include patients with HPV-positive cervical or anal cancers undergoing definitive RT or chemoradiotherapy. The study will enroll 110 participants, stratified into two groups based on post-treatment ctDNA results: 68 ctDNA-negative patients for serial ctDNA monitoring and 16 ctDNA-positive patients for Phase II immunotherapy intervention. In this phase, patients will receive Pembrolizumab 200mg IV every 3 weeks for twelve months. Endpoints: The primary endpoint for the non-interventional phase is the sensitivity and specificity of the HPV ctDNA test compared to commercial ctDNA tests and imaging. Secondary outcome is cost-effectiveness of HPV ctDNA as a follow-up tool. For the interventional phase, the primary endpoint is the 6-month disease progression rate in ctDNA-positive patients receiving immunotherapy. Secondary outcomes include recurrence rates, and survival outcomes. Clinical trial information: NCT06640283 .

High-risk human papillomavirus (HPV) is the etiological agent of nearly all cervical cancers, with HPV-18 being the second most prevalent type. The HPV life cycle is closely associated with epithelial differentiation, a process governed by cellular transcription factors (TFs). During progression toward malignancy, HPV disrupts differentiation and promotes uncontrolled cell proliferation. We aimed to identify differentiation-modulated TFs linking viral transcription to differentiation, providing insights into mechanisms driving viral life cycle and pathogenesis. DNA-binding activity of 345 TFs was compared between undifferentiated and differentiated keratinocytes to identify differentiation-associated TFs. In silico analyses identified putative binding sites for these TFs within the HPV-18 long control region (LCR). Chromatin immunoprecipitation confirmed direct binding of PAX6, HMGB1, and NFE2 to the LCR, but not FOXI1. Immunohistochemistry in keratinocyte raft cultures demonstrated differentiation-dependent expression patterns for all four TFs. Functional assays revealed that each TF is capable of modulate HPV-18 early promoter activity, with effects varying by differentiation status. Notably, expression of these TFs was disrupted by the viral oncoproteins E6 and E7, underscoring a mechanism by which HPV alters host differentiation. These findings identify novel differentiation-linked regulators of HPV-18 transcription and highlight host targets exploited by the virus in its life cycle and pathogenesis.

OBJECTIVE:In a prospective, randomized phase II study, exploratory data analysis was conducted to evaluate angiogenesis-associated plasma protein levels in patients with locally advanced cervical cancer METHODS: Participants were divided into two groups: Group A received neoadjuvant cisplatin and gemcitabine treatment (NAC) followed by chemoradiation with cisplatin and brachytherapy (CRT), while Group B received only CRT. Plasma samples were collected from patients in Group A at three time points: baseline, after NAC, and after CRT. Group B patients had samples taken at two time points: baseline and after CRT. The study analyzed an angiogenesis-associated panel of plasma proteins, including angiopoietin-2, G-CSF, endothelin-1, FGF-1, FGF-2, follistatin, IL-8, HGF, EGF, HB-EGF, PLGF, VEGF-A, VEGF-C, and VEGF- D. Receiver Operating Characteristic (ROC) analyses assessed the predictive value of baseline biomarkers for 12, 24, and 36-month survival outcomes. Additionally, Principal Component Analysis (PCA) was applied to post-CRT biomarker changes to identify coordinated modulation patterns. PCA was based on normalized delta values and eigenvector loadings, enabling identification of biomarkers aligned with Progression-Free Survival (PFS) and Overall Survival (OS). RESULTS:Significant differences were observed in the levels of HB-EGF, IL-8, PLGF, and VEGF-C between Groups A and B following CRT. Additionally, angiopoietin-2 levels showed a significant increase in Group B only. NAC treatment in Group A appeared to downregulate IL-8. CRT induced significant changes in HB-EGF, IL-8, PLGF, and VEGF-C levels in both groups. Patients in Group B demonstrated improved PFS and OS compared to those in Group A. Despite these differences in survival outcomes, the authors observed no significant intergroup differences in the tested biomarkers after completion of CRT. ROC analysis of baseline angiogenesis biomarkers demonstrated limited predictive sensitivity for survival outcomes. However, PCA of biomarker changes following CRT highlighted VEGF-A, HB-EGF, and angiopoietin-2 as variables associated with PFS and OS. CONCLUSION:Baseline biomarker levels were not predictive of long-term outcomes. In contrast, CRT alone modulated key angiogenic biomarkers, and post-CRT biomarker changes were associated with improved survival. Such biomarker alterations were not observed following NAC, which was not associated with clinical benefit in this study. These findings underscore the value of dynamic biomarker evaluation and highlight how treatment strategies differentially impact biomarker profiles in advanced cervical cancer.

Objective: This study assesses the associations between dendritic cells, HPV 6 and 11, and Recurrent Respiratory Papillomatosis (RRP) aggressiveness. Methods: The Derkay score was calculated using information obtained from the medical records. Biopsies from 36 patients with juvenile RRP (JRRP) and 43 adult RRP (ARRP) patients were analyzed under light microscopy, and their clinical data were collected. Immunohistochemical analysis using antibodies against CD83, CD1a, Factor XIIIa, and S100 was performed, and inflammatory cells were quantified. Data obtained were analyzed using the chi-squared test, in addition to the Mann–Whitney and Z tests for two proportions, considering a confidence interval of 95% and p < 0.05 as statistically significant. Results: A higher quantity of S100 was identified in the epithelium (p < 0.001) and in the conjunctive tissue (p = 0.027) among the ARRP cases, while CD83 (p = 0.025) and Factor XIIIa (p = 0.018), both in the epithelium, were identified among the JRRP cases. We observed significant association between a higher quantity of CD83 in the epithelium in the juvenile group with a low Derkay index (p = 0.034) and with HPV 6 (p = 0.039). Conclusions: An increased quantity of dendritic cells is present in individuals diagnosed with RRP, regardless of age, and this may be related to the lower Derkay index, regardless of the HPV type detected.

Oral HPV DNA and circulating tumor (ct) HPV DNA in plasma were evaluated as potential biomarkers for HPV-associated head and neck cancer (HNC). Samples from HNC patients (n = 132), including 23 oropharyngeal cancers (OPC), and non-HNC controls (n = 10) were analyzed. HPV status was determined using a multiplex bead-based test (E7-MPG) applied to formalin-fixed paraffin-embedded (FFPE) tissues (n = 90), plasma (n = 141), gargle samples (n = 141), and oral swabs (n = 142). HPV DNA was detected in 25.8% of HNC tissues, 12% of plasma samples, 20.6% of gargles and 7% of oral swabs with HPV16 as the most prevalent genotype. Among OPC cases, HPV16 DNA was found in 71.4% of FFPE samples. High concordance was observed between paired OPC tissues and plasma (91.3%) or gargles (95.2%), with moderate concordance for oral swabs (59.1%). Gargle samples alone demonstrated a 100% detection rate for HPV16-positive OPC, regardless of the cT stage, outperforming plasma (86.7%). Combined oral gargle and plasma analyses detected all HPV-positive OPC cases (7/7) at the early cT1 stage. These findings highlight the limited involvement of HPV in non-oropharyngeal HNC compared to OPC, and support gargle and plasma samples as minimally invasive diagnostic tools for detecting HPV-associated OPC.

This study aimed to assess HPV16 variant period prevalence and sequential acquisition at genital and anal sites in men from the USA and Latin America, using data from the HPV Infection in Men (HIM) Study cohort. Among 635 men with genital and 115 with anal HPV16, 53 men tested positive for any HPV16 variant at both sites, and HPV16 A1 was most prevalent at both sites, more so at the anal canal. Sequential acquisition of HPV16 A1 occurred in 30.4% (genital to anal) and 29.4% (anal to genital). These findings offer insight into HPV16 variant distribution across anatomic sites in men.

IntroductionCervical cancer, predominantly caused by persistent high-risk Human Papillomavirus (HPV) infection, remains a significant public health issue in developing countries. Identifying prognostic markers for patients with precursor lesions could improve clinical management and outcomes. MethodsWe performed a comparative gene expression analysis between low- and high-grade cervical intraepithelial lesions, focusing on genes associated with inflammation and oxidative stress. STAT1 (Signal Transducer and Activator of Transcription 1) and UCP2 (Uncoupling Protein 2), known for their roles in other cancers, were selected for further validation. Immunodetection techniques were applied in both monolayer and organotypic cultures to assess the regulation of these proteins by HPV oncoproteins. Additionally, immunohistochemistry was conducted on two patient groups: those with precursor lesions plus cancer, and those with cervical cancer only. ResultsIn cultured HaCaT cells transduced with E6/E7 HPV oncogenes, expression levels of STAT1 and UCP2 changed, especially in organotypic cultures. In patient samples, both UCP2 and STAT1 levels increased with the severity of cervical precursor lesions, and their expressions showed a strong correlation. Notably, nuclear STAT1 expression, indicative of protein activation, was rare in cancer samples but correlated with poor prognosis. In contrast, positive UCP2 expression was linked to improved survival rates and reduced recurrence. DiscussionOur findings demonstrate that HPV oncoproteins modulate the expression of UCP2 and STAT1. These proteins may serve as valuable prognostic markers for patients with precursor cervical lesions, and UCP2 expression, in particular, could be beneficial for predicting outcomes in cervical cancer patients.

Head and neck squamous cell carcinoma (SCC) predisposing factors include smoking and alcohol consumption. However, other agents have been investigated, including viruses. We aimed to investigate the presence of DNA of four different types of human polyomavirus (HPyV) in the oral cavity and oropharyngeal SCC samples from an oncology center in Brazil and evaluate the association between HPyV detection and clinical and sociodemographic characteristics. Sixty fresh frozen samples from three different anatomical sites (tongue, floor of the mouth, and oropharynx, 20 samples for each region) were retrospectively selected. Data from medical records such as age, sex, alcohol consumption, smoking, tumor staging and death in less than 5 years of diagnosis were collected. DNA was extracted for the identification of MCPyV, BKPyV, JCPyV, and TSPyV using PCR followed by Sanger sequencing of positive samples. The identity of the generated DNA sequences was confirmed by alignment reference sequences. The investigation of the presence of HPyV DNA showed positivity of 5% for MCPyV (n = 3), 0% for both BKPyV or TSPyV, and 60% for JCPyV (n = 36). No association was found between the positivity of any HPyV in samples with any clinical or sociodemographic characteristics of the patients, nor with a certain anatomical site, except for the association between death in less than 5 years after diagnosis and positivity for JCPyV (p = 0.009). Positivity for HPyV in oral cavity and oropharyngeal SCC was low for MCPyV, high for JCPyV and null for BKPyV and TSPyV. Further studies should be carried out to better understand the high prevalence of JCPyV found in oral cavity and oropharyngeal SCC.

Supplementary Table S2. Pairwise, between-genus co-occurrence of selected HPV types in the Ludwig-McGill Cohort Study (n=503)

Human papillomaviruses (HPV) of the genus Betapapillomavirus can infect both cutaneous and mucosal sites, but research on their natural history at mucosal sites remains scarce. We examined the risk factors and co-detection patterns of HPVs of the Betapapillomavirus and Alphapapillomavirus genera in cervical samples of the Ludwig-McGill cohort study. We assessed a subset of 505 women from the Ludwig-McGill cohort study from São Paulo, Brazil. Cervical samples over the first year of follow-up were tested for DNA of over 40 alphapapillomavirus types and 43 betapapillomavirus types using a type-specific multiplex genotyping polymerase chain reaction assay. We assessed the risk factors for prevalent and incident betapapillomavirus type detection, and whether types were detected more frequently together than expected assuming independence using permutation tests, logistic regression, and Cox regression. We observed significant within-genus clustering but not cross-genus clustering. Multiple betapapillomavirus types were co-detected in the same sample 2.24 (95% confidence interval [CI]: 1.65-3.29) times more frequently than expected. Conversely, co-detections of alphapapillomavirus and betapapillomavirus types in the same sample occurred only 0.64 (95% CI: 0.51-0.83) times as often as expected under independence. In prospective analyses, positivity to one HPV genus was associated with a nonsignificant lower incidence of detection of types in the other genus. Lifetime number of sex partners and new sex partner acquisition were associated with lower risks of prevalent and incident betapapillomavirus detection. Betapapillomaviruses are commonly found in the cervicovaginal tract. Results suggest potentially different mechanisms of transmission for betapapillomavirus genital infections other than vaginal sex.