The incidence of human papillomavirus (HPV) -associated oropharyngeal tumors is steadily increasing therefore, information about the prevalence of oral HPV and its risk factors is very important for future screening and early diagnosis of the disease. This thesis addresses three topics. The first topic is to evaluate the prevalence of oral HPV in a healthy population and to investigate risk factors for oral HPV infection, given that these data are almost completely absent in Central Europe. A statistically significantly higher rate of positivity (8.8%) of oral HPV infection was found in the group of older unvaccinated probands than in younger partially vaccinated volunteers (2.0%). The seropositivity rate of anamnestic HPV antibodies was comparable in both groups. None of the analyzed risk factors were significantly associated with oral HPV positivity. The second topic of the thesis is the dynamics of HPV specific antibodies in patients with head and neck cancer and their prognostic significance. In patients with cervical cancer, a decrease in HPV E6 / E7-specific antibodies is associated with a better prognosis. Another goal of the dissertation was to assess the importance of anamnestic antibodies and antibodies against oncoproteins E6 and E7 in long-term follow-up 2-14 years after the end of...

Indiana University-Purdue University Indianapolis (IUPUI) / Human papillomaviruses (HPVs) are non-enveloped, circular double-stranded DNA viruses that infect basal keratinocytes of stratified squamous epithelia. High-risk HPV (HR-HPV) infection causes nearly all cervical cancers and an increasing number of head and neck cancers. While prophylactic vaccinations have reduced the incidence of HPV infection and attributable cancers, currently there is no cure for pre-existing HPV infection. As such, HPV remains a global health threat and a better understanding of HPV biology remains of significant medical importance for identification of novel therapeutic targets. The multi-subunit structural maintenance of chromosomes 5/6 complex (SMC5/6) is comprised of SMC5, SMC6 and NSE1-4. SMC5/6 is essential for homologous recombination DNA repair and reportedly functions as an antiviral factor during hepatitis B and herpes simplex-1 viral infections. Intriguingly, SMC5/6 has been found to associate with HR-HPV E2 proteins, which are multifunctional transcription factors essential to regulation of viral replication and transcription. The function of SMC5/6 associations with E2, as well as its role during HR-HPV infection remain unclear and we explored this question in the context of HR-HPV- 31. SMC6 interacted with HPV-31 E2 and co-immunoprecipitation of SMC6/E2 complexes required the E2 transactivation domain, inferring SMC6 association is limited to the full-length E2 isoform. Depletion of SMC6 and NSE3 increased HPV replication and transcription in keratinocytes stably maintaining episomal HPV-31, suggesting that the SMC5/6 complex represses these processes. Neither SMC6 nor NSE3 co-IP the viral E1 DNA helicase alone or E1/E2 complexes but the association of SMC6 with E2 was reduced in the presence of E1, indicating that SMC6 competes with E1 for E2 binding. This infers that SMC6 repression of the viral replicative program may involve inhibiting initiation of viral replication by disrupting E2 interactions with E1. Chromatin immunoprecipitation determined that SMC6 is present on episomal HPV-31 genomes, alluding to a possible role for SMC5/6 in modifying the chromatin state of viral DNA. Taken together, these findings describe a novel function for SMC5/6 as a repressor of the HPV-31 replicative program.

SUMMARYHuman papillomaviruses (HPV) are small, non-enveloped viruses with a dsDNA genome. About 40 HPV types infect the mucosa and skin of the anogenital tract, and they are further subdivided into low-risk HPV (LR-HPV) and high-risk HPV (HR-HPV). Long-term, persistent infections with the latter type could result in the development of cervical cancer. Cervical cancer is the second most prevalent cancer worldwide, and the third leading cause of cancer-related deaths in women. HR-HPV DNA is frequently found integrated in cervical carcinoma tissue. Thus, HR-HPV integration is considered to be mechanistically linked to virus-promoted malignancy. Several complex and sometimes inter-linked signal transduction pathways, collectively termed as the DNA Damage Response (DDR), act to sense damage incurred to DNA and ultimately promote its repair. The central players of the DDR are three proteins of the phosphatidylinositol 3-kinase-like protein kinase (PIKK) family, ataxia telangiectasia mutated (ATM), ataxia telangiectasia and Rad3-related (ATR) and DNA protein kinase catalytic subunit (DNA-PKcs). Also crucial to the DDR are members of the poly(ADP)ribose polymerase family (PARP). Interactions between histones modified by PARP enzymes, the PARP enzymes themselves and the X-ray repair cross-complementing protein (XRCC-1), are important for efficient DNA repair. Reversible acetylation of histones plays a key role in the regulation of gene expression and in the DDR. The acetylation of histones, as well as that of numerous other cellular proteins, is carried out by histone acetyl transferases (HAT). This enzymatic process exists in a tightly-regulated equilibrium with lysine deacetylation, which is catalyzed by histone deacetylases (HDAC). HDAC modulate the expression and/or function of key proteins implicated in cancer. In addition, many HDAC are overexpressed in most cancers. Various small-molecule inhibitors of HDAC have been developed. HDAC inhibitors (HDACi) have been shown to promote cancer cell cycle arrest, differentiation and apoptosis. In addition, HDACi are well-documented for their synergistic or additive effects on DNA-damaging agents used in cancer therapy. In spite of a growing body of research however, the cellular and molecular mechanisms behind this phenomenon have not yet been fully uncovered. We evaluated the impact of several pan-HDACi and HDACi analogues on the extrachromosomal and genomically-integrated HPV-16 LCR-driven transcription in various cell lines. Using a classical reporter construct strategy, we report that the pan-HDACi Trichostatin A (TSA), valproate (VPA) and sodium butyrate (NaB) are potent inducers of transcription from the HPV-16 long control region (LCR) in its extrachromosomal form. This effect of HDACi was at least in part mediated by three binding sites of members of the AP-1 family of transcription factors within the viral LCR. HDACi analogues 1 through 6 did not modulate extrachromosomal HPV-16 LCR transcription in a statistically-significant way. / Doctorat en Sciences biomédicales et pharmaceutiques (Pharmacie) / info:eu-repo/semantics/nonPublished

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Forskning om stickrädda barn i elevhälsovården bedöms som otillräcklig, men betydelsefull då många vaccinationer sker i skolan. Syftet var att undersöka flickors upplevelser kring HPV- vaccination i elevhälsovården. Uppsatsstudien hade en mixad metod design och studien var baserad på enkätsvar från 14 flickor i årskurs fem och sex. Det kvantitativa resultatet visade att hälften av flickorna skattade sig som rädda i samband med HPV-vaccination och av dem skattade sig 42 % som livrädda. Upplevd smärta och rädslan för eventuell smärta var de faktorer flickorna uppgav skapade rädsla. Att ha en förälder och/eller en kompis med sig vid vaccinationen var enligt flickorna de viktigaste strategierna för att kunna minska deras rädsla. Det fanns en positiv korrelation mellan flickornas skattade rädsla och hur de upplevde att de inte kunde koncentrera sig på skolarbetet under hela veckan innan vaccination (r2=0,761; p=0,002). Det kvalitativa resultatet visade att flickorna upplevde starka fysiska och psykiska reaktioner såsom gråt, illamående, panik och stress, orsakade av rädsla inför, under och efter vaccinationen. Då elevhälsvårdens övergripande mål handlar om att förebygga både fysisk och psykisk ohälsa för barn och ungdomar är det viktigt att belysa problematiken med stickrädsla. Sammanfattningsvis behövs vidare forskning genomföras. Detta för att bättre kunna förstå hur omfattande stickrädsla är bland skolelever och för att sätta elevernas perspektiv i fokus. / Research on children with fear of needles in the student health service is estimated as inadequate but important because many vaccinations take place at school. The aim was to examine girls’ experiences of HPV-vaccination in student health services. The study had mixed method design and the study was based on survey responses from 14 girls in the fifth and sixth grade. The quantitative results showed that half of the girls estimated themselves as frightened during HPV-vaccination, and 42 % rated themselves as terrified. The percieved pain and fear of eventuall pain were the factors in the girls' experiences of HPV- vaccination that caused fear. Having a parent and/or a friend present as support are the two most common strategies for reducing the fear during the HPV-vaccination, according to the girls. There was a positive correlation between the girls' estimated fear and their percieved ability to not be able to concentrate on schoolwork during the week prior to vaccination (r2= 0.761; p = 0.002). The qualitative results showed that before, during and after vaccination, strong physical and psychological reactions such as crying, nausea, panic and stress were caused by fear. The student health services overall goal is about preventing both physical and mental health of children and adolescents and therefor it is important to highlight the problem of fear of needles. Further research is therefor needed to better understand the extent of fear of needles among schoolchildren and focus on their perspective.

Background: Human Papilloma Virus (HPV) is the recognized main reason for developing cervical cancer. HPV vaccine given to females is the most effective prevention. Purpose: To investigate knowledge and beliefs about HPV, cervical cancer and HPV vaccine among young Thai females in north-eastern part of Thailand. Further, to discover potential differences between those stating having knowledge about HPV and cervical cancer (group SHK), and those stating not having knowledge about HPV and cervical cancer (group SNHK). Method: A cross-sectional survey using a questionnaire about knowledge and beliefs of HPV where 221 young Thai females, aged 18-21, participated. Orem’s self-care theory was used as theoretical framework. Result: Less than 50 % of the participants knew about visible signs and symptoms of HPV infection. However, over 70 % had knowledge regarding HPV´s relation to sexual activity. Internet was the greatest source of information about HPV. Participants had positive belief towards the vaccine and more than 95% wished to get vaccinated. Group SHK had more knowledge then group SNHK with significant difference in seven out of fourteen knowledge items, and showed more positive beliefs with significant difference in six out of sixteen belief statements. Conclusion: The overall level of knowledge about HPV and cervical cancer was insufficient. However, this did not affect the participant’s beliefs in the subject negatively. Health care should provide viable internet sites with information about HPV to ensure that young Thai females get requisites, enabling self-care on preventing HPV infections by vaccination. / Bakgrund: Humant Papillom Virus (HPV) är den erkänt främsta orsaken till livmoderhalscancer. Vaccinering av unga kvinnor är den erkänt mest effektiva preventionen. Syfte: Att undersöka kunskap och åsikter om HPV, livmoderhalscancer och HPV vaccin bland unga thailändska kvinnor i nordöstra Thailand. Vidare, att undersöka om det fanns några skillnader mellan dem som säger sig ha kunskap om HPV och livmoderhalscancer (grupp SHK) och de som säger sig inte ha någon kunskap om HPV och livmoderhalscancer (grupp SNHK). Metod: En tvärsnittsstudie med ett frågeformulär om kunskap och åsikter om HPV som 221 unga thailändska kvinnor, i åldern 18-21, besvarade. Dorotea Orems omvårdnadsteori användes som teoretisk ram. Resultat: Mindre än 50 % av deltagarna hade kunskap om symtom av en HPV infektion. Över 70 % hade kunskap om HPV och dess relation till sexuell aktivitet. Största källan för information om HPV var internet. Deltagarna hade positiva åsikter inför vaccinet, mer än 95 % skulle vilja vaccinera sig. Grupp SHK hade mer kunskap än grupp SNHK med signifikant skillnad i sju av fjorton kunskapsämnen, och visade mer positiva åsikter med signifikant skillnad i sex av sexton påståenden rörande åsikter. Slutsats: Nivån av kunskap rörande HPV och livmoderhalscancer är otillräcklig, men det påverkar inte unga thailändska kvinnors åsikter om HPV vaccin i negativ riktning. Hälso- och sjukvården bör erbjuda korrekta och trovärdiga websidor med information om HPV för att ge unga thailändska kvinnor de förutsättningar som krävs för egenvård i prevention av HPV infektion genom vaccinering.

Ο καρκίνος του τραχήλου της μήτρας (ΚΤΜ), αποτελεί παγκοσμίως έναν από τους συχνότερα εμφανιζόμενους καρκίνους στο γυναικείο πληθυσμό. Ο ΚΤΜ επιφέρει σημαντική ψυχολογική, κοινωνική και οικονομική επιβάρυνση στους ασθενείς και στα συστήματα υγείας. Η κυριότερη αιτία εμφάνισης της νόσου συσχετίζεται με τη μόλυνση από τον ιό των ανθρώπινων θηλωμάτων – HPV (Human papilloma virus). O HPV συγκαταλέγεται στους πιο συχνά σεξουαλικ ώςμεταδιδόμενους ιούς. Ένα από τα μεγαλυτερα βήματα της δημόσιας υγείας για την προφύλαξη του πληθυσμού έναντι του HPV και κατ’ επέκταση του καρκίνου του τραχήλου της μήτρας, είναι η εισαγωγή του εμβολίου στο Εθνικό Πρόγραμμα Εμβολιασμού. Στα παρακάτω κεφάλαια (θεωρητικό μέρος) γίνεται εκτενής περιγραφή της ιστοπαθολογίας του ΚΤΜ και της βασικής βιολογίας του HPV, καθώς επίσης και του μηχανισμού δράσης του εμβολίου έναντι του ιού. Εν συνεχεία, στο ειδικό μέρος της παρούσας διπλωματικής εργασίας παρατίθενται στοιχεία που αφορούν την ευαισθητοποίηση και την αποδοσχή του πληθυσμού για τον προληπτικό εμβολιασμό έναντι του ιού HPV. / -

Gimdos kaklelio vėžys pasaulyje yra antroji, o Lietuvoje ketvirtoji pagal dažnumą moterų onkologinė liga. Pagrindinė gimdos kaklelio vėžio priežastis yra žmogaus papilomos viruso (ŽPV) infekcija gimdos kaklelyje. Vakarų šalyse ikivėžiniams pokyčiams ar vėžinėms ląstelės nustatyti yra naudojamas citologija paremtas Papanicolaou testas (Pap) ir ŽPV DNR testas. Šiuo metu naudojami ŽPV diagnostiniai testai remiasi molekulinės biologijos tyrimo metodais. Tyrimui buvo pasirinktas polimerazės grandininės reakcijos (PGR) metodas, o ne plačiai naudojamas JAV bei Europos kokybės sertifikatus turintis Hybrid Capture 2 (HC2) metodas. PGR yra ypač jautrus, greitas, komerciškai prieinamas metodas, kuriam reikalingas nedidelis tiriamosios medžiagos kiekis, ir kuriuo galima tiksliai identifikuoti ŽPV tipus. HC2 metodas nenustato ŽPV genotipų, galimi klaidingai neigiami rezultatai, nes nėra vidinės kontrolės, o dėl kryžminių reakcijų galimi klaidingai teigiami rezultatai. Naujausi literatūros šaltiniai teigia, kad PGR turi daug perspektyvų ateityje ir manoma, kad ŽPV nustatymo standartas bus 14 DR-ŽPV tyrimas sujungtas su ŽPV-16 ir ŽPV-18 tipų nustatymu. Todėl siekiant ŽPV tyrimams pasirinkti optimaliausią iš Lietuvoje siūlomų komercinių rinkinių Valstybiniame patologijos centre buvo atliekamas šis tyrimas. Įvertinus tyrimų rezultatus, tolesniems tyrimams pasirinktas „Seeplex® HPV4A ACE Screening“ ŽPV nustatymo rinkinys. / Cervical cancer is the second oncological disease in women in the world, and the fourth one in Lithuania. The main cause of the cervical cancer is Human papillomavirus (HPV) infection in the cervix uteri. In order to detect precancerous changes or cancer cells, a Papanicolaou (Pap) test based on cytology and HPV DNA test are used in the Western world. Currently used HPV diagnostic tests are based on molecular biology testing methods. The polymerase chain reaction (PCR) method, not the extensively used Hybrid Capture 2 (HC2) method awarded the certificate of quality by both the USA and Europe, was chosen. PCR is a particularly sensitive, fast, and commercially available method that needs a small quantity of research material and can identify the types of HPV. HC2 method does not detect HPV genotypes, false-negative results are possible because there is no internal control, and due to cross reactions, false-positive results are possible. The newest written sources state that PCR will have much potential in the future and it is thought that HPV detection standard will become 14 HR-HPV test combined with the detection of the types of HPV-16 and HPV-18. Therefore, in order to chose the optimum one for HPV tests from the commercial kits offered in Lithuania, this investigation was carried out in the National Centre of Pathology. Four different HPV diagnostic kits were used for the research. Having evaluated the research results, „Seeplex® HPV4A ACE Screening“ HPV diagnostic kit was... [to full text]

Les infections à papillomavirus humains à haut-risque (HPV-HR) sont responsables de 100% des cancers cervico-utérins et de 50% des carcinomes de l'oropharynx. Les infections du col utérin sont généralement transitoires et bénignes. Cependant, en cas d'infection persistante, elles peuvent s'accompagner d'une progression vers des lésions (pré)cancéreuses du col utérin. Les facteurs viraux qui favorisent la persistance ou la clairance virale sont encore mal connus. Dans une première partie, nous avons étudié la méthylation des régions 3'L1 et LCR dans des frottis cervico-utérins HPV16+ / cytologie normale, prélevés chez 37 femmes qui présenteront soit une infection transitoire soit une infection persistante avec progression vers une lésion précancéreuse. Nous avons montré que certains îlots CpG présentaient des taux de méthylation différents en fonction de l'évolution ultérieure de l'infection HPV et pourraient être utilisés comme marqueurs prédictifs.Les cancers de l'oropharynx associés à une infection à HPV-HR présentent un pronostic plus favorable que ceux associés à une intoxication alcoolo-tabagique. Les mécanismes responsables de ce meilleur pronostic sont encore mal compris. Dans une seconde partie, nous avons caractérisé le statut HPV dans 202 cas de cancers de l'oropharynx. Au sein des 32 cancers présentant une infection HPV16 active (avec expression des ARNm E6/E7), nous avons étudié la méthylation des régions 3'L1 et LCR et l'intégration du génome viral. Nous avons montré que les niveaux de méthylation de certains sites CpG, notamment des sites de liaison à la protéine virale E2 (E2BS3 et E2BS4) sont fortement méthylés dans les formes épisomales et mixtes mais sont non méthylés dans les formes intégrées pures.Enfin dans une troisième partie, nous avons étudié des marqueurs de transition épithélio-mésenchymateuse (TEM) dans des modèles cellulaires et des cancers oropharyngés, en fonction du statut HPV. Une moindre TEM, connue pour être impliquée de façon importante dans les phénomènes d'invasion tumorale, pourrait expliquer le meilleur pronostic associé aux cancers HPV-induits. Nous avons montré que la vimentine, marqueur le plus représentatif de l'acquisition de capacités migratoires et invasives, est plus exprimée dans les cellules tumorales HPV positives que dans les cellules HPV négatives. Le pronostic différentiel observé entre les 2 étiologies des cancers oropharyngés ne serait donc pas corrélé à la TEM.Mots-clés : Cancer, HPV, col utérin, oropharynx, biomarqueurs, méthylation, intégration, TEM. / High-risk human papillomavirus (HR-HPV) infections are responsible for 100% of cervical cancers and 50% of oropharyngeal cancers. Cervical infections are usually transient and benign. Persistent infections may however progress and lead to cervical (pre)cancerous lesions. Viral factors contributing to persistence or clearance are poorly understood. In a fist part, we studied methylation of 3'L1 and LCR regions in HPV16+ / normal cytology cervical smears, taken in 37 women who will present either a transient infection, or a persistent infection with progression to a precancerous lesion. We demonstrated that some CpG sites harbored differential methylation rates in relation with later outcome of HPV infection and may be used as predictive biomarkers.HR-HPV related oropharyngeal cancers present a better prognostic that their alcohol-and-tobacco-induced counterparts. Mechanisms responsible for this better prognostic are mainly unknown. In a second part, we characterized the HPV status in 202 oropharyngeal cancers. In the 32 cases presenting with HPV16 active infections (expressing E6/E7 mRNA), we studied 3'L1 and LCR methylation and integration of viral genome. We showed that methylation rates of some CpG sites, especially in E2 binding sites (E2BS3 & 4), were strongly methylated in episomal and mixed forms but were unmethylated in purely integrated forms.In a third part, we studied epithelial-to-mesenchymal-transition (EMT) markers in cellular models and oropharyngeal cancers, according to HPV status. A lesser EMT, known as an important phenomenon implicated in tumor invasion, could explain the better prognostic associated with HPV-induced cancers. We showed that vimentin, the more representative marker for migration and invasion ability acquisition is more expressed in HPV+ tumor cells than in HPV-negative cells. So the differential prognosis observed between the 2 oropharyngeal cancer etiologies may not be linked to EMT.Key-words: Cancer, HPV, cervical cancer, oropharynx, biomarkers, methylation, integration, EMT.