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34 pages, 2998 KiB  
Article
An Assessment of Young Adults’ Awareness and Knowledge Related to the Human Papillomavirus (HPV), Oropharyngeal Cancer, and the HPV Vaccine
by Eric N. Davis and Philip C. Doyle
Cancers 2025, 17(3), 344; https://doi.org/10.3390/cancers17030344 - 21 Jan 2025
Abstract
Background/Objectives: The human papillomavirus (HPV) is a prevalent sexually transmitted infection that is a known cause of morbidities such as genital warts and cancers of the cervix, anus, and oropharynx. Non-cervical HPV-related cancers have been a developing problem in North America, increasing in [...] Read more.
Background/Objectives: The human papillomavirus (HPV) is a prevalent sexually transmitted infection that is a known cause of morbidities such as genital warts and cancers of the cervix, anus, and oropharynx. Non-cervical HPV-related cancers have been a developing problem in North America, increasing in incidence by up to 225% in some instances over a span of two decades. Methods: This study investigated levels of awareness and knowledge of HPV, oropharyngeal cancer (OPC), and the HPV vaccine using a self-administered web-based survey designed specifically for this research. University students (n = 1005) aged 18–30 completed a 42-item questionnaire that included demographic information, awareness questions, and a series of “true/false/I don’t know” knowledge questions. Results: The data gathered revealed that participants had relatively high levels of awareness. However, many respondents had significant gaps in their knowledge of HPV, OPC, and the HPV vaccine. Collectively, the data indicate that awareness and knowledge of HPV and the value of vaccination may place younger individuals at risk for HPV-related infections. Conclusions: Although a relatively high level of awareness concerning HPV was observed, the gaps in knowledge suggest that further efforts are necessary to educate young adults. While all risk factors cannot be reduced, the present data may guide future efforts directed toward better education on HPV and related health concerns and associated risks. Full article
(This article belongs to the Special Issue Advances in Virus-Associated Head and Neck Cancer)
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<p>Age of participants. Note: 13 participants did not specify their age.</p>
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<p>Participant vaccination status.</p>
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<p>HPV knowledge sources.</p>
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<p>Perceived level of concern about HPV infection.</p>
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<p>Self-perceived HPV knowledge levels.</p>
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<p>Distribution of responses to HPV knowledge questions.</p>
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<p>Perceived level of concern about developing OPC.</p>
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<p>Self-perceived OPC knowledge levels.</p>
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<p>OPC knowledge questions and responses.</p>
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<p>HPV vaccine knowledge questions and responses.</p>
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20 pages, 564 KiB  
Article
Genetic Variants of Interleukin-8 and Interleukin-16 and Their Association with Cervical Cancer Risk
by Rafał Watrowski, Eva Schuster, Stefan Polterauer, Toon Van Gorp, Gerda Hofstetter, Michael B. Fischer, Sven Mahner, Robert Zeillinger and Eva Obermayr
Life 2025, 15(2), 135; https://doi.org/10.3390/life15020135 - 21 Jan 2025
Viewed by 163
Abstract
Background: Cervical cancer (CC) is the fourth most common cancer diagnosis in women worldwide. Infection with high-risk human papillomavirus (HPV) is a critical but not determinative condition for CC development, as several co-factors modulate the progression of HPV-associated cervical lesions. Interleukin-8 (IL-8) and [...] Read more.
Background: Cervical cancer (CC) is the fourth most common cancer diagnosis in women worldwide. Infection with high-risk human papillomavirus (HPV) is a critical but not determinative condition for CC development, as several co-factors modulate the progression of HPV-associated cervical lesions. Interleukin-8 (IL-8) and Interleukin-16 (IL-16) are chemokine-like interleukins involved in the pathogenesis of various cancers. Singular studies in Asian populations have suggested a potential role of IL-8 rs4073 (−251 A>T) and IL-16 rs1131445 (3′UTR T>C) in cervical carcinogenesis. Methods: A case-control study was conducted in a European cohort of 339 women, including 126 CC patients and 213 controls. Four common IL-8 SNPs, rs4073 (−251 A>T), rs2227306 (+781 C>T), rs1126647 (+2767 A>T), and rs2227543 (+1633 C>T), and four IL-16 polymorphism, rs4778889 (−295 T>C), rs11556218 (3441 T>G), rs4072111 (1300 C>T), and rs1131445 (3′UTR T>C), were assessed using RFLP-PCR and analyzed under seven inheritance models. Subgroup analyses were stratified by menopausal status (age threshold 51 years), disease stage, and histological subtype. Results: IL-16 rs4072111 was significantly associated with an increased CC risk in premenopausal women in the co-dominant (p = 0.038), dominant (p = 0.022), and heterozygote (p = 0.045) models, identifying the T allele as the risk allele (OR 2.31, CI95% 1.17–4.56; p = 0.017). In women aged over 51, IL-16 rs4778889 was associated with CC in the heterozygote (p = 0.048) and overdominant (p = 0.042) models but not in the co-dominant model (p = 0.092). None of the analyzed SNPs significantly increased CC risk in the entire cohort. Specifically, neither IL-16 rs1131445 nor IL-8 rs4073, previously reported as risk factors in Asian populations, were associated with CC risk in this European cohort. Conclusions: These findings highlight the role of age stage in immunity and cancer susceptibility, suggest that IL-8 and IL-16 SNPs may function differently in cervical carcinogenesis compared with other cancers, and emphasize the importance of ethnic background in cancer risk, warranting further research. Full article
(This article belongs to the Section Medical Research)
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<p>Age distribution and density between CC cases (at first diagnosis) and controls.</p>
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12 pages, 597 KiB  
Article
Racial and Ethnic Disparities in Human Papillomavirus Vaccination Among US Born and Foreign-Born Adults Aged 18 to 26 Years in the United States
by Itunu Sokale, Jane Montealegre, Ann O. Amuta, Abiodun Oluyomi and Aaron P. Thrift
Vaccines 2025, 13(2), 98; https://doi.org/10.3390/vaccines13020098 (registering DOI) - 21 Jan 2025
Viewed by 151
Abstract
Background/Objectives: Human papillomavirus (HPV) is linked to multiple cancers that can be prevented through vaccination. While the optimal age for vaccination is in childhood and adolescence, vaccination recommendations include adults through age 26 who missed childhood/adolescent vaccination. There are limited data about disparities [...] Read more.
Background/Objectives: Human papillomavirus (HPV) is linked to multiple cancers that can be prevented through vaccination. While the optimal age for vaccination is in childhood and adolescence, vaccination recommendations include adults through age 26 who missed childhood/adolescent vaccination. There are limited data about disparities among adults eligible for catch-up HPV vaccination. We conducted a comprehensive examination of HPV vaccination among US young adults, disaggregating the group by race/ethnicity and nativity status to identify subgroups that may require additional interventions. Methods: We analyzed 2019 and 2022 data of individuals aged 18–26 years from the National Health Interview Survey. Generalized linear models using Poisson regression with log link were used to examine the receipt of 1+ dose of HPV vaccine, race/ethnicity, and nativity (i.e., US- versus foreign-born) status. Results: The overall receipt of 1+ doses of HPV vaccine was 47.5%. The vaccination rate among the US-born group was 49.7% versus 31.9% among the foreign-born group with an adjusted prevalence ratio (APR) of 0.72; (95% CI, 0.62–0.82). Foreign-born non-Hispanic (NH) Black individuals (APR 0.31; 95% CI, 0.13–0.70) were less likely to be vaccinated against HPV than foreign-born NH White individuals, while US-born NH Asians (APR 1.27; 95% CI, 1.09–1.48) had a higher prevalence of the vaccination than the US-born NH White group. Additionally, foreign-born NH Asian (APR 0.60; 95% CI, 0.46–0.77), NH Black (APR 0.27; 95% CI, 0.12–0.61), and Hispanic (APR 0.76; 95% CI, 0.60–0.97) populations were less likely to be vaccinated than their respective US-born counterparts. Conclusion: Profound HPV vaccination inequalities exist among US young adults with particularly low vaccine coverage among racially and ethnically minoritized immigrant populations. Full article
(This article belongs to the Section Human Vaccines and Public Health)
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<p>Weighted HPV vaccination rates among US young adults aged 18–26 years, by nativity and race/ethnicity. Abbreviations: HPV, human papillomavirus; NH, non-Hispanic. Race/ethnicity groups are mutually exclusive, and Hispanic/Latinos could be of any race. Other includes American Indian/Alaska Native, Asian, Native Hawaiian/Pacific Islander, and multiple races. Respondents were considered foreign-born if they reported that they were not born in the United States or any of its territories.</p>
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<p>Weighted HPV vaccination rates among racial–ethnic groups of US young adults aged 18–26 years stratified by nativity. Abbreviations: HPV, human papillomavirus; NH, non-Hispanic; Race/ethnicity groups are mutually exclusive, and Hispanic/Latinos could be of any race; Other includes American Indian/Alaska Native; Asian, Native Hawaiian/Pacific Islander, and multiple races. Respondents were considered foreign-born if they reported that they were not born in the United States or any of its territories.</p>
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16 pages, 706 KiB  
Article
Impact of HPV Catch-Up Vaccination on High-Grade Cervical Lesions (CIN2+) Among Women Aged 26–30 in Northern Norway
by Amanda Sørensen Jørgensen, Gunnar Skov Simonsen and Sveinung Wergeland Sørbye
Vaccines 2025, 13(1), 96; https://doi.org/10.3390/vaccines13010096 - 20 Jan 2025
Viewed by 199
Abstract
Background/Objectives: Human papillomavirus (HPV) is the primary cause of high-grade cervical lesions and cervical cancer worldwide. In Norway, HPV vaccination was introduced in 2009 for seventh-grade girls and extended through a catch-up program from 2016 to 2019 for women born between 1991 [...] Read more.
Background/Objectives: Human papillomavirus (HPV) is the primary cause of high-grade cervical lesions and cervical cancer worldwide. In Norway, HPV vaccination was introduced in 2009 for seventh-grade girls and extended through a catch-up program from 2016 to 2019 for women born between 1991 and 1996. This study evaluates the impact of the catch-up vaccination program on the incidence of HPV and high-grade cervical lesions in Troms and Finnmark. Methods: We analyzed data from 40,617 women aged 26 to 30 who underwent cervical screening between 2009 and 2023 in Troms and Finnmark, including 1850 women with high-grade cervical lesions (CIN2+) on biopsy. Using linear regression, we assessed trends in high-grade lesion incidence per 1000 screened women and the association between vaccination status and HPV-16/18 incidence. Results: Between 2017 and 2023, the incidence of high-grade cervical lesions significantly decreased: CIN2+ decreased by 33.4%, and CIN3+ decreased by 63.4%. Significant reductions in HPV-16/18-associated high-grade cervical lesions were observed among vaccinated women, with the proportion of CIN2+ cases due to HPV-16 and 18 decreasing from 56.8% in 2017 to 40.7% in 2023, reflecting a 55.8% reduction in the absolute number of cases caused by these high-risk HPV types. Comparing unvaccinated women aged 25–26 in 2016 and vaccinated women in 2023, HPV-16 incidence decreased from 5.1% to 0.1%, and HPV-18 incidence decreased from 3.3% to 0.0%. Conclusions: The catch-up vaccination program significantly reduced the incidence of HPV-16/18 and high-grade cervical lesions in Troms and Finnmark, even with the lower vaccination coverage observed in the catch-up program. These findings demonstrate the effectiveness of HPV vaccination programs in reducing HPV infections and associated cervical lesions. Full article
27 pages, 811 KiB  
Review
Advancing the Fight Against Cervical Cancer: The Promise of Therapeutic HPV Vaccines
by Qian Zheng, Misi He, Zejia Mao, Yue Huang, Xiuying Li, Ling Long, Mingfang Guo and Dongling Zou
Vaccines 2025, 13(1), 92; https://doi.org/10.3390/vaccines13010092 - 19 Jan 2025
Viewed by 387
Abstract
Human papillomavirus (HPV) is a major global health issue and is recognized as the leading cause of cervical cancer. While prophylactic vaccination programs have led to substantial reductions in both HPV infection rates and cervical cancer incidence, considerable burdens of HPV-related diseases persist, [...] Read more.
Human papillomavirus (HPV) is a major global health issue and is recognized as the leading cause of cervical cancer. While prophylactic vaccination programs have led to substantial reductions in both HPV infection rates and cervical cancer incidence, considerable burdens of HPV-related diseases persist, particularly in developing countries with inadequate vaccine coverage and uptake. The development of therapeutic vaccines for HPV represents an emerging strategy that has the potential to bolster the fight against cervical cancer. Unlike current prophylactic vaccines designed to prevent new infections, therapeutic vaccines aim to eradicate or treat existing HPV infections, as well as HPV-associated precancers and cancers. This review focuses on clinical studies involving therapeutic HPV vaccines for cervical cancer, specifically in three key areas: the treatment of cervical intraepithelial neoplasia; the treatment of cervical cancer in combination with or without chemotherapy, radiotherapy, or immune checkpoint inhibitors; and the role of prophylaxis following completion of treatment. Currently, there are no approved therapeutic HPV vaccines worldwide; however, active progress is being made in clinical research and development using multiple platforms such as peptides, proteins, DNA, RNA, bacterial vectors, viral vectors, and cell-based, each offering relative advantages and limitations for delivering HPV antigens and generating targeted immune responses. We outline preferred vaccine parameters, including indications, target populations, safety considerations, efficacy considerations, and immunization strategies. Lastly, we emphasize that therapeutic vaccines for HPV that are currently under development could be an important new tool in fighting against cervical cancer. Full article
(This article belongs to the Special Issue Vaccine Strategies for HPV-Related Cancers)
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<p>Mechanism of therapeutic HPV vaccines. Traditional vaccines such as DNA vaccines, RNA vaccines, peptide vaccines, protein vaccines, and vector vaccines can deliver target antigens to antigen-presenting cells (APCs), which then present the target antigen epitopes on their surface through major histocompatibility complex class I (MHC I) and MHC II. With the assistance of CD4+ helper T cells, activated HPV antigen-specific CD8+ T cells produce cytotoxic T lymphocytes (CTLs), which kill cells infected with HPV [<a href="#B22-vaccines-13-00092" class="html-bibr">22</a>,<a href="#B23-vaccines-13-00092" class="html-bibr">23</a>]. HPV Tumor Infiltrating Lymphocyte (HPV-TIL) vaccines [<a href="#B31-vaccines-13-00092" class="html-bibr">31</a>] involve the process of separating and culturing TILs from the patient’s tumor tissue, and then screening lymphocytes that are reactive to HPV E6/E7 antigens for reinfusion back into the patient. Isolating T cells or other immune cells from the patient’s blood, transfecting and amplifying these cells ex vivo with HPV E6/E7, and preparing them into T cell receptor-engineered T cell (TCR-T) vaccines [<a href="#B27-vaccines-13-00092" class="html-bibr">27</a>] and autologous peripheral blood mononuclear cell (PBMC) vaccines [<a href="#B28-vaccines-13-00092" class="html-bibr">28</a>], followed by reinfusion back into the patient.</p>
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19 pages, 614 KiB  
Review
A Canadian Perspective on Systemic Therapy for Recurrent or Metastatic Nasopharyngeal Carcinoma
by Anna Spreafico, Eric Winquist, Cheryl Ho, Brian O’Sullivan, Nathaniel Bouganim, Neil Chua, Sarah Doucette, Lillian L. Siu and Desiree Hao
Curr. Oncol. 2025, 32(1), 48; https://doi.org/10.3390/curroncol32010048 - 17 Jan 2025
Viewed by 453
Abstract
Although the majority of patients with nasopharyngeal carcinoma (NPC) present with early-stage or locoregional disease that can be treated with definitive radiotherapy, approximately 20% of patients experience disease recurrence, and 15% present with metastatic disease that is not amenable to curative therapy. Management [...] Read more.
Although the majority of patients with nasopharyngeal carcinoma (NPC) present with early-stage or locoregional disease that can be treated with definitive radiotherapy, approximately 20% of patients experience disease recurrence, and 15% present with metastatic disease that is not amenable to curative therapy. Management of patients with recurrent or metastatic (r/m) NPC who are not candidates for local salvage therapy is challenging in Canada, as there is uncertainty in extrapolating evidence that is largely generated from Southeast China to non-endemic regions such as Canada. Currently, treatment options in Canada are limited to chemotherapy regimens that can only achieve short-term response and prolongation of survival. The addition of anti-PD-1 monoclonal antibodies to chemotherapy has been shown to extend progression-free survival in recurrent r/m NPC compared to chemotherapy alone; however, approval of PD-1 inhibitors in Canada has lagged behind other jurisdictions where NPC is non-endemic. This paper reviews the current systemic treatment landscape for r/m NPC in Canada, highlights unmet treatment needs for patients who are not candidates for curative therapy, and discusses the challenges and opportunities that lie in emerging therapies. Full article
(This article belongs to the Section Head and Neck Oncology)
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<p>Treatment algorithm for r/m NPC and treatment access in Canada, the United States, Europe, and China. <sup>a</sup> Includes single agents/combinations of paclitaxel, docetaxel, 5-FU, capecitabine, gemcitabine, irinotecan, vinorelbine, cisplatin, oxaliplatin, carboplatin, ifosfamide, doxorubicin, methotrexate, and/or cetuximab, depending on region and preference. <sup>b</sup> Access to clinical trials specifically for patients with NPC is more limited in Canada, the United States, and Europe. <sup>c</sup> Listed as a recommended regimen in the National Comprehensive Cancer Network guideline. Not approved by the FDA for this specific indication. EBV, Epstein-Barr virus; HNSCC, head and neck squamous cell carcinoma; HPV, human papillomavirus; IMRT, intensity-modulated radiation therapy; NPC, nasopharyngeal carcinoma; r/m, recurrent/metastatic; WHO, World Health Organization.</p>
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16 pages, 232 KiB  
Article
Provider Preference, Logistical Challenges, or Vaccine Hesitancy? Analyzing Parental Decision-Making in School Vaccination Programs: A Qualitative Study in Sydney, Australia
by Leigh McIndoe, Alexandra Young, Cristyn Davies, Cassandra Vujovich-Dunn, Stephanie Kean, Michelle Dives and Vicky Sheppeard
Vaccines 2025, 13(1), 83; https://doi.org/10.3390/vaccines13010083 - 17 Jan 2025
Viewed by 284
Abstract
Background: School-based immunization programs are crucial for equitable vaccine coverage, yet their success depends on parental consent processes. This study investigates patterns of vaccine decision-making within Australia’s school-based immunization program, specifically focusing on human papillomavirus (HPV) and diphtheria-tetanus-pertussis (dTpa) vaccines offered free to [...] Read more.
Background: School-based immunization programs are crucial for equitable vaccine coverage, yet their success depends on parental consent processes. This study investigates patterns of vaccine decision-making within Australia’s school-based immunization program, specifically focusing on human papillomavirus (HPV) and diphtheria-tetanus-pertussis (dTpa) vaccines offered free to adolescents aged 12–13. Methods: This qualitative study was conducted in the South Eastern Sydney Local Health District (2022–2023). Semi-structured interviews were held with school staff (n = 11) across government, Catholic, and independent schools, parents whose children were not vaccinated at school (n = 11) and a focus group with public health unit staff (n = 5). Data were analyzed to identify key barriers and patterns in vaccine decision-making. Results: Analysis revealed three distinct groups of parents whose children were not vaccinated through the school program: (1) those favoring general practitioners for vaccination, driven by trust in medical providers and a preference for personalized care; (2) those intending to consent but facing logistical barriers, including communication breakdowns and online consent challenges; and (3) vaccine-hesitant parents, particularly regarding HPV vaccination, influenced by safety concerns and misinformation. These findings demonstrate that non-participation in school vaccination programs should not be automatically equated with vaccine hesitancy. Conclusions: Tailored interventions are necessary for addressing vaccine non-participation. Recommendations include strengthening collaboration with general practitioners, streamlining consent processes and providing targeted education to counter misinformation. This study provides valuable insights into social determinants of vaccine acceptance and offers actionable strategies for improving vaccine uptake in school-based programs. Full article
(This article belongs to the Special Issue Advancing the Science on Vaccine Hesitancy to Inform Interventions)
14 pages, 1688 KiB  
Article
Vaginal Dysbiosis in Infertility: A Comparative Analysis Between Women with Primary and Secondary Infertility
by Iliana Alejandra Cortés-Ortíz, Gustavo Acosta-Altamirano, Rafael Nambo-Venegas, Jesús Alejandro Pineda-Migranas, Oscar Giovanni Ríos-Hernández, Eduardo García-Moncada, Alejandra Yareth Bonilla-Cortés, Mónica Sierra-Martínez and Juan Carlos Bravata-Alcántara
Microorganisms 2025, 13(1), 188; https://doi.org/10.3390/microorganisms13010188 - 17 Jan 2025
Viewed by 451
Abstract
Infertility, both primary and secondary, is strongly influenced by microbiological factors, with the vaginal microbiota playing a key role in reproductive health. Objective: The aim of this study was to characterize the vaginal microbiota of 136 Mexican women diagnosed with infertility—primary (n = [...] Read more.
Infertility, both primary and secondary, is strongly influenced by microbiological factors, with the vaginal microbiota playing a key role in reproductive health. Objective: The aim of this study was to characterize the vaginal microbiota of 136 Mexican women diagnosed with infertility—primary (n = 58) and secondary (n = 78)—by evaluating the presence of pathogenic bacterial species and their associations with infertility conditions. Methods: Samples were obtained through cervical swabs, and microorganism identification was performed using qPCR techniques. Results: Analysis revealed a positive correlation between increased age and the likelihood of primary infertility, as well as a negative correlation with secondary infertility. Significant differences in microbial composition were also observed between the two infertility groups. Lactobacillus crispatus and Lactobacillus gasseri were dominant in women with primary infertility, in addition to a high prevalence of Gardnerella vaginalis and Fannyhessea vaginae. Additionally, correlations were found between the presence of human papillomavirus (HPV) and sexually transmitted bacteria, as well as Gardnerella vaginalis. Conclusion: Our findings suggest that the composition of the vaginal microbiota may play a decisive role in infertility, highlighting the need for personalized therapeutic strategies based on microbial profiles. Full article
(This article belongs to the Section Medical Microbiology)
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<p>The histogram shows the age distribution of patients with primary infertility (blue bar) and secondary infertility (orange bar). Ages range from 18 to 48 years. For primary infertility, a peak is observed at 37 years, with patients predominantly distributed between 25 and 40 years and lower representation at the extremes of the range. For secondary infertility, the distribution is more uniform, with a peak around 29 years, consistent frequencies between 26 and 40 years, and some dispersion toward older ages (up to 45 years).</p>
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<p>The vaginal microbiome community state types. (<b>A</b>) Cluster formation analysis. Five clusters were generated from all variables across all samples using the K-Means method. (<b>B</b>) Key variables impacting cluster formation. A Random Forest analysis was conducted to determine the variables that contribute to the groupings, with the most significant variables being age, microbial community type (CST), the presence of sexually transmitted bacteria, type of infertility, and infection with the human papillomavirus (HPV).</p>
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<p>Relationship between age and type of infertility. Logistic regression models were constructed to evaluate the potential relationships between these factors. A positive relationship can be observed between increasing age and the likelihood of primary infertility.</p>
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<p>Correlations between bacterial species and viruses in different community state types. Principal component analysis (PCA) of community group types. The graph suggests correlations between the variables: smaller angles indicate a positive correlation, larger angles indicate a negative correlation, and 90° angles indicate no correlation between the characteristics. The distance between points represents their similarity: close points reflect similar profiles, while points that are distant exhibit dissimilar profiles.</p>
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17 pages, 10117 KiB  
Article
A Conserved Di-Lysine Motif in the E2 Transactivation Domain Regulates MmuPV1 Replication and Disease Progression
by Jessica Gonzalez, Marsha DeSmet and Elliot J. Androphy
Pathogens 2025, 14(1), 84; https://doi.org/10.3390/pathogens14010084 - 16 Jan 2025
Viewed by 292
Abstract
The papillomavirus E2 protein regulates the transcription, replication, and segregation of viral episomes within the host cell. A multitude of post-translational modifications have been identified which control E2 functions. A highly conserved di-lysine motif within the transactivation domain (TAD) has been shown to [...] Read more.
The papillomavirus E2 protein regulates the transcription, replication, and segregation of viral episomes within the host cell. A multitude of post-translational modifications have been identified which control E2 functions. A highly conserved di-lysine motif within the transactivation domain (TAD) has been shown to regulate the normal functions of the E2 proteins of BPV-1, SfPV1, HPV-16, and HPV-31. This motif is similarly conserved in the E2 of the murine papillomavirus, MmuPV1. Using site-directed mutagenesis, we show that the first lysine (K) residue within the motif, K112, is absolutely required for E2-mediated transcription and transient replication in vitro. Furthermore, mutation of the second lysine residue, K113, to the potential acetyl-lysine mimic glutamine (Q) abrogated E2 transcription and decreased transient replication in vitro, while the acetylation defective arginine (R) mutant remained functional. Both K113 mutants were able to induce wart formation in vivo, though disease progression appeared to be delayed in the K113Q group. These findings suggest that acetylation of K113 may act as a mechanism for repressing MmuPV1 E2 activity. Full article
(This article belongs to the Section Viral Pathogens)
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<p>The TAD di-lysine motif is highly conserved. (<b>A</b>) Multiple sequence alignment of BPV-1, SfPV1, HPV-16, HPV-31, and MmuPV1 E2 protein sequences. The di-lysine motif (highlighted in yellow) is conserved across papillomavirus types. (<b>B</b>) Structure alignment between the solved BPV-1 TAD (cyan, PDB 2JEU) and the predicted MmuPV1 TAD (dark blue). The first lysine of the motif is depicted in red, and the second lysine is depicted in magenta. Structure prediction was performed with I-TASSER-MTD [<a href="#B25-pathogens-14-00084" class="html-bibr">25</a>]; alignment and visualization were performed in PyMOL.</p>
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<p>The first lysine in the di-lysine motif is critical for MmuPV1 E2 function. (<b>A</b>) Mutants of the first lysine, K112, fail to activate transcription from the pGL2-4xE2BS-Luc reporter. (<b>B</b>) Transcriptional activity is not restored by over-expression of the K112 mutants. WT E2 activates transcription in a dose-dependent manner. pCI-V5-E2 plasmid amounts were as follows: 50 ng (1×), 250 ng (5×), 500 ng (10×). (<b>C</b>) Immunoblot of cell lysates from panel C shows that K112 mutants are expressed in a dose-dependent manner. Mouse anti-V5 (1:3000), mouse anti-α-tubulin (1:10,000). (<b>D</b>) Neither K112Q nor K112R can support transient replication in the presence of E1. WT E2 demonstrates robust transient replication when co-expressed with WT E1. (<b>E</b>) Immunofluorescence was performed in HEK293TT cells transfected with expression vectors for WT or K112 mutant V5-E2. Both mutants are present within the nucleus, indicating that mutation of K112 does not interfere with nuclear localization of MmuPV1 E2. Mouse anti-V5 (1:1500), AlexaFluor 488 anti-mouse (1:3000). Nucleus is stained with DAPI. <span class="html-italic">n</span> = 3, *** <span class="html-italic">p</span> &lt; 0.001, paired <span class="html-italic">t</span>-test. Scale bar represents 10 μM.</p>
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<p>K113Q is transcriptionally defective. (<b>A</b>) K113Q is unable to activate transcription to a significant level, even when over-expressed ten-fold. K113R and WT E2 can activate transcription in a dose-dependent manner when over-expressed. * <span class="html-italic">p</span> &lt; 0.05, Dunnett’s test. E2 vector amounts are titrated as 1× (50 ng), 5× (250 ng), and 10× (500 ng). (<b>B</b>) The data in panel A are presented and analyzed for comparison against WT E2. Data are grouped by expression level to demonstrate that K113R transcription is significantly diminished compared to WT E2. <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, paired <span class="html-italic">t</span>-test. (<b>C</b>) Immunoblot of whole cell lysate. Both K113 mutants display decreased protein levels compared to WT E2 at the 1× expression level, but protein levels increase when the mutants are over-expressed. Mouse anti-V5 (1:3000), mouse anti-GAPDH (1:8000).</p>
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<p>K113Q is deficient for transient replication. C33-A cells were transfected with mFLori, pRLuc, WT or K113 mutant pCI-V5-E2, and WT or TTL pCI-myc-E1. Luminescence was measured 72 h post-transfection. (<b>A</b>) Both mutants can transiently replicate compared to samples without E1. (<b>B</b>) Comparisons were made between WT and K113 mutant samples containing E1. K113Q showed a significant defect compared to WT E2. K113R consistently replicated to lower levels than WT E2, but this difference was not significant. <span class="html-italic">n</span> = 3, *** <span class="html-italic">p</span> &lt; 0.001, paired <span class="html-italic">t</span>-test.</p>
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<p>Mutation of the di-lysine motif does not interfere with nuclear localization. CV-1 cells were transfected with 3 μg of WT or mutant pCI-V5-E2. Cells were fixed 48 h post-transfection and stained for V5-E2 (Rabbit anti-V5, 1:1200; AlexaFluor 594 1:3000) and mounted with DAPI. Slides were analyzed for localization of E2 (red) within the nucleus (DAPI). Cells transfected with an mCherry expression vector were used as a control demonstrating diffuse localization. All di-lysine mutants were localized to the nucleus. Scale bar represents 10 μM.</p>
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<p>K113Q shows diminished binding to E1. HEK293TT cells were transfected with expression vectors for myc-E1 and WT, K113Q, or K113R V5-E2. Lysates were co-immunoprecipitated with anti-myc magnetic beads and protein complexes were separated by SDS-PAGE. WT (lane 3) and K113R (lane 5) immunoprecipitated with E1, while K113Q (lane 4) shows decreased binding. Immunoblotted with rabbit anti-Myc tag (1:3000) and rabbit anti-V5 (1:3000).</p>
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<p>Both K113Q and K113R can induce wart formation in vivo. (<b>A</b>) Timeline of disease progression. Athymic hairless mice were injected intradermally with approximately 20 μg of circular mutant genomes and monitored weekly for the development of cutaneous lesions. K113Q lesion development lags behind K113R throughout the experiment. (<b>B</b>) Muzzle and tail lesions at the time of sacrifice, 4.5 months post-injection. K113R mice demonstrate florid lesion growth on the muzzle indicative of secondary infection, while only one K113Q mouse has developed a lesion on the muzzle. (<b>C</b>) Sequence alignment between DNA isolated from collected lesions and the MmuPV1 reference genome. K113 is highlighted in yellow, confirming that the lesions do contain the indicated E2 mutation.</p>
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<p>K113Q and K113R induce dysplasia in mice. Lesions were collected from the tail and muzzle and formalin fixed for H&amp;E staining. The top panels are tissue sections without visible lesions and display normal histologic features. The lesions collected from both anatomic sites demonstrate histologic changes characteristic of PV-induced disease, including hyperkeratosis, parakeratosis, and koilocytic change (Images were taken at 20× magnification).</p>
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24 pages, 1140 KiB  
Article
The Relationship Between Cervicovaginal Infection, Human Papillomavirus Infection and Cervical Intraepithelial Neoplasia in Romanian Women
by Anca Daniela Brăila, Cristian-Viorel Poalelungi, Cristina-Crenguţa Albu, Constantin Marian Damian, Laurențiu Mihai Dȋră, Andreea-Mariana Bănățeanu and Claudia Florina Bogdan-Andreescu
Diseases 2025, 13(1), 18; https://doi.org/10.3390/diseases13010018 - 16 Jan 2025
Viewed by 445
Abstract
Cervical intraepithelial neoplasia (CIN) is a premalignant cervical condition closely linked to persistent high-risk HPV infection, a major risk factor for cervical cancer. This study aims to investigate the relationship between cervicovaginal infections, HPV infection, and CIN development in 94 Romanian women with [...] Read more.
Cervical intraepithelial neoplasia (CIN) is a premalignant cervical condition closely linked to persistent high-risk HPV infection, a major risk factor for cervical cancer. This study aims to investigate the relationship between cervicovaginal infections, HPV infection, and CIN development in 94 Romanian women with cervical lesions. Comprehensive assessments included HPV genotyping, cytology, colposcopy, and histopathology. In 53.20% of cases, vaginal infections were identified, with Candida albicans most frequently associated with HPV. Histopathology revealed 48.94% low-grade CIN, 42.55% high-grade CIN, and 8.51% invasive carcinoma. There was a strong correlation between high-risk HPV types (especially HPV 16 and 18), colposcopic findings, histopathology, and age. This study emphasizes the mutual effect of cervicovaginal infections and HPV infection in increasing the risk of developing CIN and cervical cancer among Romanian women. Persistent infection with high-risk HPV types, particularly HPV 16 and 18, has been confirmed as a primary driver of CIN and cervical cancer progression. Full article
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<p>Distribution of cases in the study cohort.</p>
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12 pages, 279 KiB  
Article
Epstein–Barr Virus (EBV) and Human Papilloma Virus (HPV) in Gastric Cancers, with Special Reference to Gastric Cancer at a Young Age—A Pilot Study in Poland
by Marek Mazurek, Małgorzata Jaros, Anna M. Gliwa, Monika Z. Sitarz, Ewa Dudzińska, Krzysztof Zinkiewicz and Robert Sitarz
Int. J. Mol. Sci. 2025, 26(2), 711; https://doi.org/10.3390/ijms26020711 - 16 Jan 2025
Viewed by 354
Abstract
Gastric cancer (GC) is one of the most common cancers in the world. It is a multi-factorial disease influenced by both genetic and environmental factors such as diet, obesity, radiation exposure, and infectious agents. Viral infections usually lead to chronic inflammation, which can [...] Read more.
Gastric cancer (GC) is one of the most common cancers in the world. It is a multi-factorial disease influenced by both genetic and environmental factors such as diet, obesity, radiation exposure, and infectious agents. Viral infections usually lead to chronic inflammation, which can initiate the development of cancers. To date, only a few studies have been published about Epstein–Barr virus (EBV) and human papillomavirus (HPV) infections in the context of the development of GC. In particular, research on the development of cancer among people under 45 years of age, including the impacts of EBV and HPV, is rare, and clear results have not been obtained. The aim of this study was to analyze the frequency of occurrence of EBV and HPV in GC, particularly in early-onset gastric cancer (EOGC). Tissue material from 135 patients with GC, including 84 men and 51 women, was examined. RT-PCR was performed to detect EBV, and PCR was performed to detect HPV. There were no significant impacts of EBV and HPV infections on any subtype of GC. There was also no statistically significant dependence of gender and location of the tumor on any subtype of GC. Further research on the impacts of infectious agents such as EBV and HPV on GC should be conducted using larger populations. Full article
(This article belongs to the Special Issue Molecular Diagnosis and Latest Treatment for Gastric Cancer)
14 pages, 455 KiB  
Article
Implementation of Valid HPV Diagnostics for the Early Detection of Cervical Cancer in Molecular Pathology: HPV 3.5 LCD-Array (Chipron GmbH) vs. PapilloCheck® (Greiner Bio-One GmbH) vs. VisionArray® (ZytoVision GmbH)
by Jan Jeroch, Melanie Winter, Anna Bieber, Agnes Boger, Christina Schmitt, Silvana Ebner, Morva Tahmasbi Rad, Henning Reis and Peter. J. Wild
J. Mol. Pathol. 2025, 6(1), 3; https://doi.org/10.3390/jmp6010003 - 15 Jan 2025
Viewed by 361
Abstract
The occurrence of cervical cancer is often linked to a previous infection with a human papillomavirus (HPV). In order to detect HPV infections in cervical smears, a broad range of tests can be used. This study compares the two hybridisation-based DNA-microarray systems “HPV [...] Read more.
The occurrence of cervical cancer is often linked to a previous infection with a human papillomavirus (HPV). In order to detect HPV infections in cervical smears, a broad range of tests can be used. This study compares the two hybridisation-based DNA-microarray systems “HPV 3.5 LCD-Array” (Chipron GmbH) and “PapilloCheck®” (Greiner Bio-One GmbH), based on their ability to detect and differentiate HPV infections in 42 different cervical smears. PapilloCheck® can detect and individually identify 24 HPV types, whereas the 3.5 LCD-Array can differentiate among 32 HPV genotypes. However, both systems include all 13 high-risk (HR)-classified types. With Chipron having already stopped the production of the 3.5 LCD-Array test, quite a few laboratories are confronted with the need to establish a new HPV testing method. The two methods were found to have a high agreement regarding the clinical significance of the detected HR HPV types. Discrepant cases were additionally validated with the help of a third test (VisionArray® HPV, ZytoVision GmbH). The results of the VisionArray® test corresponded rather well with the results of the 3.5 LCD-Array. Full article
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<p>Number of HPV types detected by the four different extraction/detection methods. HPV types 51, 53, and 42 were the only ones detected by the 3.5 LCD-Array and simultaneously by all extraction/detection methods. The combination of oCheck<sup>®</sup> and PapilloCheck<sup>®</sup> was the only one detecting the HR HPV types 16 and 39 and the possibly cancerogenic HPV type 70. oCheck<sup>®</sup> and VisionArray<sup>®</sup> was the only pairing that could detect HPV 6.</p>
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17 pages, 2844 KiB  
Article
Developing an Effective Therapeutic HPV Vaccine to Eradicate Large Tumors by Genetically Fusing Xcl1 and Incorporating IL-9 as Molecular Adjuvants
by Zhongjie Sun, Zhongyan Wu and Xuncheng Su
Vaccines 2025, 13(1), 49; https://doi.org/10.3390/vaccines13010049 - 9 Jan 2025
Viewed by 457
Abstract
Background: Human papillomavirus (HPV) is a prevalent infection affecting both men and women, leading to various cytological lesions. Therapeutic vaccines mount a HPV-specific CD8+ cytotoxic T lymphocyte response, thus clearing HPV-infected cells. However, no therapeutic vaccines targeting HPV are currently approved for clinical [...] Read more.
Background: Human papillomavirus (HPV) is a prevalent infection affecting both men and women, leading to various cytological lesions. Therapeutic vaccines mount a HPV-specific CD8+ cytotoxic T lymphocyte response, thus clearing HPV-infected cells. However, no therapeutic vaccines targeting HPV are currently approved for clinical treatment due to limited efficacy. Our goal is to develop a vaccine that can effectively eliminate tumors caused by HPV. Methods: We genetically fused the chemokine XCL1 with the E6 and E7 proteins of HPV16 to target cDC1 and enhance the vaccine-induced cytotoxic T cell response, ultimately developing a DNA vaccine. Additionally, we screened various interleukins and identified IL-9 as an effective molecular adjuvant for our DNA vaccine. Results: The fusion of Xcl1 significantly improved the quantity and quality of the specific CD8+ T cells. The fusion of Xcl1 also increased immune cell infiltration into the tumor microenvironment. The inclusion of IL-9 significantly elevated the vaccine-induced specific T cell response and enhanced anti-tumor efficacy. IL-9 promotes the formation of central memory T cells. Conclusions: the fusion of Xcl1 and the use of IL-9 as a molecular adjuvant represent promising strategies for vaccine development. Full article
(This article belongs to the Section DNA and mRNA Vaccines)
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<p>Construction and analysis of HPV16 therapeutic DNA vaccine fused with chemokine Xcl1. (<b>A</b>) The structural prediction diagram of the E6E7 protein and the Xcl1-E6E7 fusion protein. In the diagram, dark blue indicates Xcl1, red represents the linker, bright blue denotes E6, and light purple signifies E7. (<b>B</b>) Following the transfection of HEK293T cells for 24 h, Western blot analysis was conducted to assess the expression levels of the E6E7 and Xcl1-E6E7 DNA vaccines, alongside the protein levels of P53 and Rb. (<b>C</b>) Flow cytometry was employed to determine the proportion of CD11c+CD8+ DC cells bound to E6E7 and Xcl1-E6E7 proteins. The left panel presents a representative flow cytometry chart, while the right panel displays the corresponding statistical analysis (N = 3, mean ± SEM).</p>
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<p>Efficacy analysis of E6E7 and Xcl1-E6E7 plasmid DNA vaccines. (<b>A</b>) E7-Specific CD8+ T cell response. Peripheral blood samples were collected on days 9, 14, 21, and 28 following the administration of 5 µg and 25 µg doses of E6E7 and Xcl1-E6E7 vaccines. The E7-specific CD8+ T cell responses were dynamically analyzed (N = 5, mean ± SEM). (<b>B</b>) The DNA vaccine injection frequency patterns. (<b>C</b>) The graph illustrates the dynamics of E7-specific T cell levels over time under various injection frequency regimens (N = 5, mean ± SEM). (<b>D</b>,<b>E</b>) After 14 days later administering 25 µg (<b>D</b>) and 5 µg (<b>E</b>) doses of the E6E7 and Xcl1-E6E7 vaccines, tumor cells were inoculated to assess tumor growth, with growth curves plotted accordingly (N = 5, mean ± SEM). (<b>F</b>) Tumor growth post inoculation and vaccination. Four days following tumor cell inoculation, 25 µg doses of the E6E7 and Xcl1-E6E7 vaccines were administered. Tumor sizes were measured and growth curves were plotted. A second tumor inoculation was conducted in mice where tumors completely regressed (N = 5, mean ± SEM). (<b>G</b>) Fourteen days post vaccination in figure (<b>F</b>), flow cytometry was used to assess E7-specific CD8+ T cell levels in the peripheral blood of mice from each group (N = 5, mean ± SEM).</p>
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<p>Analysis of immune cell subpopulations within the tumor microenvironment. (<b>A</b>) Flow cytometry analysis of the proportion of E7-specific CD8+ T cells in tumors from mice treated with E6E7 or Xcl1-E6E7 vaccines. (<b>B</b>) Detection of the proportion of Granzyme B (GranB)-positive cells within the E7-specific CD8+ T cell population in each treatment group. (<b>C</b>–<b>E</b>) Flow cytometry analysis of CD45+ total white blood cells and the proportions of CD8+ and CD4+ T cells within the tumor microenvironment of mice treated with E6E7 or Xcl1-E6E7 vaccines. (<b>F</b>) Flow cytometry analysis of the proportion of regulatory T cells (Tregs) within the CD4+ T cell population. (<b>G</b>) Flow cytometry analysis of the proportion of natural killer (NK) cells relative to total white blood cells in the tumor microenvironment of mice in each group. (N = 5, mean ± SEM) for each figure.</p>
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<p>IL-9 enhances the therapeutic effect of the Xcl1-E6E7 plasmid DNA vaccine. (<b>A</b>–<b>C</b>) Mice with established subcutaneous tumors, averaging 30 mm<sup>3</sup> in volume, formed by 1 × 10<sup>5</sup> TC-1 cells per mouse, were treated with either E6E7 or Xcl1-E6E7 plasmid DNA. Tumor growth curves were plotted for individual mice across the empty vector group (<b>A</b>), E6E7 group (<b>B</b>), and Xcl1-E6E7 group (<b>C</b>). (<b>D</b>) Xcl1-E6E7 DNA vaccine was administered 4 or 7 days after TC-1 tumor cell inoculation. The levels of E7-specific CD8+ T cells in peripheral blood were dynamically monitored and plotted as a curve. (<b>E</b>) Fourteen days post immunization with the Xcl1-E6E7 plasmid DNA vaccine combined with plasmid DNA expressing IL-7, IL-9, IL-21, or IL-33, E7-specific CD8+ T cell levels were assessed in peripheral blood. (<b>F</b>,<b>G</b>) Fourteen days after immunization with plasmid DNA expressing IL-9 and IL-33 in combination with GPC3 plasmid DNA vaccines, ELISPOT analysis of mouse spleen cells was conducted (<b>G</b>), F showing representative IFN-γ spots. (<b>H</b>,<b>I</b>) The MPEC proportion of E7-specific CD8+ T cells in the spleen of Xcl1-E6E7 with and without IL-9-immunized mice was analyzed after 28 days, along with the expression of the central memory marker CD62L in MPECs. (<b>J</b>) The plots show the tumor growth curve of Xcl1-E6E7 with and without IL-9-treated mice (N = 5, mean ± SEM).</p>
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<p>Efficacy analysis of Xcl1-E6E7 + mIL-9 combined with immune checkpoint inhibitors. (<b>A</b>) Flow cytometry analysis of the proportions of CD4+ and CD8+ T cells in peripheral blood, conducted one week after the administration of anti-CD4 and anti-CD8 antibodies. (<b>B</b>) Fourteen days post immunization with Xcl1-E6E7 + mIL-9, mice were inoculated with 5 × 10<sup>5</sup> TC-1 tumor cells to observe tumor growth. Anti-CD4 and anti-CD8 antibodies were administered intraperitoneally starting one week after vaccination, with dosing twice a week (N = 5, mean ± SEM). (<b>C</b>) Mice with TC-1 xenograft tumors were treated with 10, 25, and 100 µg doses of the Xcl1-E6E7+mIL-9 vaccine. Tumor volumes in each group were measured regularly, and growth curves were plotted (N = 7, mean ± SEM). (<b>D</b>,<b>E</b>) Mice were subcutaneously inoculated with 5 × 10<sup>5</sup> TC-1 tumor cells to form tumors, then randomly assigned to treatment groups. Treatments included anti-OX40, anti-4-1BB, anti-PD-1, anti-PD-L1, and anti-CTLA-4, either alone (<b>D</b>) or in combination with Xcl1-E6E7 + mIL-9 plasmid DNA (<b>E</b>). Tumor growth was monitored, and growth curves were plotted (N = 5, mean ± SEM). (<b>F</b>) Dynamic monitoring of E7-specific CD8+ T cell levels in the peripheral blood of mice immunized with XCL1-E6E7 + mIL-9 plasmid DNA, either alone or in combination with anti-CTLA-4 antibody (N = 5, mean ± SEM).</p>
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11 pages, 677 KiB  
Communication
Beliefs and Perceptions in Attending the Cervical Screening: The COMUNISS Project Experience
by Narcisa Muresu, Illari Sechi, Mariangela Valentina Puci, Marco Dettori and Andrea Piana
Cancers 2025, 17(2), 190; https://doi.org/10.3390/cancers17020190 - 9 Jan 2025
Viewed by 429
Abstract
Background: Several studies highlighted that tailored health communication interventions improve cervical screening participation, vaccination coverage, and awareness about self-sampling benefits. The “COMUNISS” project was aimed at increasing awareness about cervical cancer prevention, identifying barriers to screening, and promoting screening uptake in under-screened women. [...] Read more.
Background: Several studies highlighted that tailored health communication interventions improve cervical screening participation, vaccination coverage, and awareness about self-sampling benefits. The “COMUNISS” project was aimed at increasing awareness about cervical cancer prevention, identifying barriers to screening, and promoting screening uptake in under-screened women. Methods: A dedicated website with a Q&A session regarding HPV-associated diseases has been set up. Participants were invited to complete a questionnaire to gather demographic information, knowledge about HPV and cervical cancer, and attitudes toward screening based on the Health Beliefs Model (HBM). Women can also require a vaginal self-sampling kit at your home to perform the HPV-DNA analysis. Results: The website registered over 1000 users over 6 months, and 256 women completed the survey. Nearly half were under-screened. The HBM revealed a high susceptibility and severity perception of diseases, regardless of screening participation, whereas older women declared a high perception of barriers. One-quarter of the women who had requested the self-collection kit returned it for the HPV-DNA testing. Conclusions: The project found significant gaps in knowledge regarding extra-cervical HPV-related cancers, interpretation of screening results, and effectiveness of self-collection. These findings highlight the need to plan targeted information campaigns to enhance awareness and adherence to screening programs. Full article
(This article belongs to the Special Issue Cervical Cancer: Screening and Treatment in 2024-2025)
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<p>Participation in screening programs by age, civil status, level of education, and occupational status.</p>
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16 pages, 2506 KiB  
Article
Evaluating Tumor Tissue Modified Viral (TTMV)-HPV DNA for the Early Detection of Anal Squamous Cell Carcinoma Recurrence
by Rafi Kabarriti, Shane Lloyd, James Jabalee, Catherine Del Vecchio Fitz, Randa Tao, Tyler Slater, Corbin Jacobs, Sean Inocencio, Michael Rutenberg, Chance Matthiesen, Kasha Neff, Gene-Fu Liu, Tiffany M. Juarez and Stanley L. Liauw
Cancers 2025, 17(2), 174; https://doi.org/10.3390/cancers17020174 - 8 Jan 2025
Viewed by 589
Abstract
Background: The incidence and mortality of anal squamous cell carcinoma (ASCC) are rising, with greater than 80% of cases linked to human papillomavirus (HPV), primarily HPV16. Post-treatment surveillance can be challenging due to the limitations of anoscopy, digital anal rectal exam (DARE), and [...] Read more.
Background: The incidence and mortality of anal squamous cell carcinoma (ASCC) are rising, with greater than 80% of cases linked to human papillomavirus (HPV), primarily HPV16. Post-treatment surveillance can be challenging due to the limitations of anoscopy, digital anal rectal exam (DARE), and imaging. Plasma tumor tissue modified viral (TTMV)-HPV DNA has shown strong sensitivity, specificity, and predictive value in detecting the recurrence of HPV-driven oropharyngeal cancer. Here, we investigate the ability of TTMV-HPV DNA for the early recurrence detection of ASCC. Methods: This retrospective clinical case series included 117 patients with HPV-driven ASCC across 7 U.S. centers, monitored with TTMV-HPV DNA during routine clinical care between March 2020 and June 2024. Physician-reported clinical data and biomarker testing data were combined to create a comprehensive, longitudinal dataset for evaluating test performance metrics. Results: Patients had a median age of 63 years and median post-diagnosis follow-up of 19 months. HPV status was primarily confirmed by TTMV-HPV DNA (52%) or p16 immunohistochemistry (39%). Of those tested for TTMV-HPV DNA pretreatment, 85% had a positive result. TTMV-HPV DNA clearance during or within three months post-treatment was associated with significantly better recurrence-free survival. The per-patient surveillance sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 82.8%, 98.4%, 96.0%, and 92.5%. Of 24 patients with a documented recurrence and a positive TTMV-HPV DNA test, the test was the first evidence of recurrence in 14 patients (58.3%), with a median lead time of 59 days (range: 10–536). TTMV-HPV DNA accurately resolved 94.3% of cases with indeterminate clinical findings. Conclusions: TTMV-HPV DNA testing provides a sensitive and specific approach for detecting patients with recurrent ASCC and resolving the status of patients with indeterminate clinical findings. Full article
(This article belongs to the Section Cancer Biomarkers)
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<p>Distribution of TTMV-HPV DNA tests by treatment stage and timing. (<b>a</b>) Bar graph showing the distribution of patients according to the number of tests received. (<b>b</b>) Bar graph displaying the proportion of post-treatment tests by time interval, with intervals defined as left-inclusive.</p>
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<p>Pretreatment score distribution. Tukey-style boxplot illustrating the distribution of positive pretreatment test scores.</p>
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<p>Post-treatment TTMV-HPV DNA score distribution and effect of positive testing on recurrence-free survival. (<b>a</b>) Tukey-style boxplot illustrating the distribution of positive post-treatment TTMV-HPV DNA scores. (<b>b</b>) Survival curve in which patients with post-treatment positive testing are separated based on whether they have positive testing during post-treatment. Kaplan–Meier method with two-sided log-rank test. CI: Confidence interval. HR: Hazard ratio.</p>
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<p>CONSORT diagram of patient inclusion, testing, and recurrence. * There were a total of 24 recurrences across 19 patients with positive TTMV-HPV DNA testing. ‡ Across 24 total recurrences, 14 were associated with early true positive testing and 10 with confirmatory true positive testing.</p>
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<p>Swimmer plots for patients with clinically confirmed recurrence or positive post-treatment TTMV-HPV DNA testing. (<b>a</b>) Early true positives (<span class="html-italic">n</span> = 14). TTMV-HPV DNA was the first sign of recurrence. (<b>b</b>) Confirmatory true positives (<span class="html-italic">n</span> = 10). Positive TTMV-HPV DNA was coincident with or following clinical recurrence detection. (<b>c</b>) False positives (<span class="html-italic">n</span> = 1). Positive TTMV-HPV DNA in the absence of clinical recurrence detection. (<b>d</b>) False negatives (<span class="html-italic">n</span> = 5). TTMV-HPV DNA was negative and occurred either after, or within three months before, clinical recurrence detection. Only patient “ac” had a biopsy-confirmed HPV-positive recurrence.</p>
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<p>Baseline TTMV-HPV DNA resolution and recurrence-free survival. (<b>a</b>) Tukey-style boxplot comparing pretreatment TTMV-HPV DNA scores for patients whose test resolves vs. those whose tests remain unresolved by three months post-treatment. (<b>b</b>) Survival curve in which patients with pretreatment positive testing are separated based on whether their test score resolves during or within three months after initial treatment. Kaplan–Meier method with two-sided log-rank test. CI: Confidence interval. HR: Hazard ratio. ns: not significant.</p>
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