Mestrado (dissertação)—Universidade de Brasília, Faculdade de Ciências da Saúde, Programa de Pós-Graduação em Saúde Coletiva, 2017. / Submitted by Gabriela Lima (gabrieladaduch@gmail.com) on 2017-12-06T09:25:57Z No. of bitstreams: 1 2017_JêniferOlivattodaSilva.pdf: 1437610 bytes, checksum: 1b331445bb55f2cb7fc2711c8ba14f26 (MD5) / Approved for entry into archive by Raquel Viana (raquelviana@bce.unb.br) on 2018-01-31T18:58:33Z (GMT) No. of bitstreams: 1 2017_JêniferOlivattodaSilva.pdf: 1437610 bytes, checksum: 1b331445bb55f2cb7fc2711c8ba14f26 (MD5) / Made available in DSpace on 2018-01-31T18:58:33Z (GMT). No. of bitstreams: 1 2017_JêniferOlivattodaSilva.pdf: 1437610 bytes, checksum: 1b331445bb55f2cb7fc2711c8ba14f26 (MD5) Previous issue date: 2018-01-31 / O Centro de Testagem e Aconselhamento (CTA) é um serviço estratégico para promoção da saúde, prevenção, diagnóstico rápido e tratamento de HIV, sífilis, hepatite B e C e, outras infecções sexualmente transmissíveis (IST). O HPV é a IST mais comum mundialmente, atuando como forte fator causal para câncer anal, de colo de útero e pênis. O objetivo principal deste estudo foi descrever as características epidemiológicas e clínicas da população adulta, masculina, diagnosticada com HPV, atendida no CTA-DF, no período de 01/01/2015 a 31/12/2016. Para contemplar este objetivo o estudo contou com dois métodos: estudo epidemiológico do tipo transversal, descritivo, com uso de dados secundários contidos no Sistema de Informação – CTA; e estudo epidemiológico do tipo coorte clínica, descritivo, com coleta de dados secundários dos prontuários da instituição. Foi identificado no Sistema de Informação do CTA (SI-CTA) a predominância do sexo masculino com 18.543 (66,12%), solteiros com 19.292 (68,79%), pardos com 12.411 (44,26%), homens que fazem sexo com mulheres 11.051 (65,69%), com a faixa etária entre 18 a 30 anos de 15.509 (55,30%) e com 8 a 11 anos de estudo 11.825 (42,17%). Os percentuais de positividade para os testes rápidos foram: 3,00% de HIV; 8,23% de sífilis; 0,10% de hepatite B e 0,39% de hepatite C. As prevalências entre coinfecções foram: 0,9% para HIV e sífilis; 0,06% para sífilis e hepatite C, 0,03% para HIV e hepatite B, 0,02% para sífilis e hepatite B e 0,01% para HIV e hepatite B. Foi verificado nos prontuários dos homens com diagnóstico de HPV a predominância da faixa etária entre 18 a 29 anos, brancos, homens que fazem sexo com mulheres, com 8 a 11 anos de estudo, com prevalência de HPV de 16,93/1.000 usuários. No que se refere à coinfecção com HPV a predominante foi HIV com 21 (7,9%) usuários. Em relação ao tipo de lesão a mais frequente foi a lesão de pênis com 209 (60,23%) usuários dos HPV positivos. No que diz respeito ao tratamento o Ácido Tricloracético (ATA 90%) foi o mais prevalente com mediana de 41 dias de tratamento e entre 3 a 5 sessões, dependendo do local da lesão. Em relação ao desfecho houve abandono de tratamento em 208 (66,24%) usuários, alta em 96 (30,57%) dos usuários e foram encaminhados para outros serviços de referência 10 (3,18%) usuários. Observou-se a importância da qualidade dos dados para que possam ser transformados em informação, conhecimento e posterior convertê-los em ações e investigações epidemiológicas. Os achados do estudo reforçam a importância de se traçar este perfil no DF, para fomentar ações de promoção da saúde, prevenção e controle de IST. Desta forma, a faixa etária da vacinação para homens e mulheres devem ser estendidas, pelas características epidemiológicas encontradas no estudo, pois o homem ativo pode ser o transmissor tanto para homens como para mulheres. / The Counseling and Testing Center (CTC) is a strategic service for health promotion, prevention, early diagnosis and treatment of sexually transmitted infections (STIs). The Human Papilloma Virus (HPV) is the largest sexual infection in occurrence worldwide, acting as a strong causal factor for anal, uterine cervix and penis cancers. The main purpose of this thesis was to describe the clinical and epidemiological characteristics of the adult, male population diagnosed with HPV, attended at the CTC in the period from January 1, 2015 to December 31, 2016. To achieve this goal the study used two methods, an epidemiological descriptive cross-sectional study, using secondary data contained in the service information system; and an descriptive epidemiological study of the clinical cohort collecting secondary data from the institution records. It was identified in the Information System the predominance of males 18.543 (66,12%), single 18.543 (66,12%), brown 12.411 (44,26%), men who have sex with women 12.411 (44,26%), between 18 and 30 years 12.411 (44,26%) and with 8 to 11 years of study 12.411 (44,26%). The percentages of positivity were: 3,00% HIV; 8.23% syphilis; 0.10% 0.39% hepatitis B and hepatitis C. The prevalence of coinfections were: 0.90% for HIV and Syphilis; 0.06% syphilis and hepatitis C, 0.03% for hepatitis B and HIV, syphilis and 0.02% for hepatitis B and 0.01% for HIV and hepatitis B. It was verified in the medical records of men with HPV the predominance of the 18 to 29 age group, white, men who have sex with women, with 8 to 11 years of study, with prevalence of HPV 16,93 / 1000 users. As regards the co-infection with HIV is the predominant HPV 21 (7.9%) users. Regarding the most common type of injury was the penis lesion 209 (60.23%) users. Regarding the treatment of HPV, trichloroacetic acid (ATA 90%) was the most prevalent median of 41 days of treatment and from 3 to 5 sessions, depending on the site of the lesion. An outcome of the treatment with an overestimated dropout rate (66.24%) and an underestimated discharge rate (30.57%) is observed, which may have had a heavy effect on the actual number of dropouts. The study indicates that the CTC is a service that can provide access to diagnosis and treatment of STIs men. The service in this way plays an important role in the interruption of the Federal District's transmission chain, since the sexually active man can be an agent of transmission of HPV for both women and men. The findings of the study reinforce the importance of drawing this profile in the DF, for the actions of health promotion, prevention and control of STIs.

Introdução: A infecção pelo Papiloma Virus Humano (HPV) vem sendo reportada como uma das doenças sexualmente transmissíveis com maior incidência na atualidade, porém a sua prevalência não é bem esclarecida em homens, principalmente devido a baixa presença de sintomas. Além disso, poucos estudos foram realizados nesta população até o momento para verificar a resposta imune pós-vacinação. As hipóteses testadas serão fundamentais para aprofundar o conhecimento da imunopatogênese, da resposta vacinal em pacientes infectados pelo HIV e colaborar no desenho e estratégias de vacinação anti-HPV na população infectada pelo HIV Objetivos: Analisar a resposta específica aos antígenos da vacina anti-HPV em homens infectados pelo HIV. Métodos: Um total de 24 pacientes infectados pelo HIV que preencheram os critérios de inclusão durante o período de coleta foram vacinados pela vacina anti-HPV bivalente em três doses nos períodos: zero, dois e seis meses. Os grupos foram divididos em: Grupo Controle (Cinco indivíduos sadios, com sorologia negativa para HIV); Grupo A (Nove pacientes com CD4 <500 celulas mm³); Grupo B (10 pacientes com CD4 >=500 celulas mm³). Foram realizados ELISA para a detecção de anticorpos Anti-HPV nos momentos pré e pós-vacinação nos grupos estudados; posteriormente realizamos nos mesmos o ensaio de cultura celular para detecção de citocinas (IFN?, IL17, TNF, IL6 e IL10) pela técnica de CBA . Resultados: Obtivemos soroconversão da primeira dose da vacina para o grupo A 55,6%, grupo B 30%, grupo controle 60%; na segunda dose obtivemos para o grupo A 88,8%, grupo B 80%, grupo controle 80%, e por final a terceira dose no grupo A 88,8%, grupo B 90%, grupo controle 100%. A citocina IL 6 (perfil TH2) demonstrou níveis mais elevados, comparados entre os grupos A, B e grupo controle (p<0.001). A partir da 3° dose da vacinação observamos baixos níveis de INF-? (perfil TH1) A e B (p<0.0006). O grupo controle apresentou produção de INF- ? quando comparado com grupos A e B (p<0.001). Conclusão: Os pacientes soropositivos e grupo controle foram respondedores a vacinação anti-HPV. Foi demonstrada uma elevada produção das citocinas entre os grupos sugerindo uma imunomodulação do grupo HIV+. Esse trabalho apresenta informações relevantes que estimulam a realização de novos estudos nessa população, avaliações de reações cruzada da vacina que pode resultar em proteção a outros tipos de HPV não presentes na vacina, além de analisar por mais tempo as titulações no soro desses pacientes. Os dados do nosso estudo podem corroborar para a vacinação nessa população, diminuindo assim o risco de uma infecção, mortalidade e morbidade das doenças causadas pelo HPV em homens. / Introduction: Infection with Human Papilloma Virus (HPV) has been reported as one of the sexually transmitted diseases with a higher incidence nowadys, but its prevalence must be clarified in men, mainly due to low presence of symptoms. Moreover, few studies have been performed in this population until now to verify the immune response post-vaccination. The hypothesis here suggested will be the key for better understanding of the immunopathogenesis, the vaccine´s response in HIV-infected patients and collaborate in the design and strategies of vaccination against HPV in HIV-infected population. Objectives: Analyze the specific response to antigens of HPV vaccine in HIV-infected men. Methods: A total of 24 HIV-infected patients who were in accordance with the inclusion criteria during the data collection period were vaccinated with anti-HPV bivalent vaccine in three period doses: zero, two and six months. The groups were distributed in: Control group (five healthy subjects with negative serology against HIV); Group A (nine subjects with CD4 <500 cells/mm³; Group B (10 subjects with CD4 >500 cells/mm³). ELISA was performed to detect the level of antibodies anti-HPV before and after vaccination in the studied cohort. Postenarly, cells of these groups were submitted in culture to verify citokynes production (IFN?, IL17, TNF, IL6 and IL10) using CBA methodology. Results: We obtained seroconversion after the first dose of anti-HPV vaccine: control group 60%, group A 55,6% and group B 30%. In the second dose: control group 80%, group A 88,8% and Group B 80%. And at last, the third dose: Control Group 100%, Group A 88,8% and group B 90%. IL 6 citokyne (TH2 response) was detected in higher level when compared Control, A and B groups (p<0.001). IFN? citokyne (TH1 response) was detect in low level only after the third dose of vaccination, showing relevance between A and B groups (p<0.0006). Additionally, higher IFN? production was detected when compared the control with A and B groups (p<0.001). Conclusion: HIV patients and controls (HIV-) were responders to anti-HPV vaccination. It was clear that an elevated cytokine production was detected between groups, suggesting immunomodulation of HIV + group. This work suggests relevant information that challenge: new studies in this population, verification of cross-reactions of the vaccine resulting in protection of other HPV types not present in this vaccine, and analyze for longer period the titers of anti-HPV antibodies in these patients. All together, our data can corroborate for vaccination in this population, thus decreasing the risk of infection, mortality and morbidity of the disease caused by HPV in men.

Cartas al Editor

Les Papillomavirus Humains (HPV) à haut risque carcinogène, dont HPV16, sont les agents étiologiques du cancer du col de l'utérus. Le génom d'HPV est composé d'un ADN double brin comprenant deux régions codantes : précoce« E » et tardive« L » et une région régulatrice non codante, la LCR. La protéine E2 se fixe au niveau des E2BS, situés dans la LCR et réprime l'expression d'E6 et d'E7. La perte d'expression d'E2 suite à l'intégration du génome virale induit une surexpression d'E6 et d'E7 qui favorisent la dégradation de p53 et de pRb. Des dinucléotides CpG étant présents au niveau des E2BS d'HPV16, nous avons détenniné si l'expression d'E6 était soumise à une régulation épigénétique. Nous avons développé une PCR HRM pour étudier le niveau de méthylation des E2BS dans les lésions précancéreuses et cancéreuses et nous avons noté la présence de CpG méthylés uniquement dans les cancers. Par ailleurs, nous avons montré que la méthylation des E2BS limite la fixation d'E2 et pern1et probablement la surexpression d'E6 et d'E7. Enfin, nous avons montré que le traitement des cellules HPV16 dérivées de cancer du col utérin pi le 5azadC, induit une diminution de l'expression d'E6. Ce mécanisme est indépendant d'E2 et nous avons prouvé que la ré-expression du miR-375, qu cible les transcrits E6/E7, est responsable de la répression d'E6 après traitement par SazadC. L'ensemble de nos résultats ont montré que l'expression des oncoprotéines d'HPV16 est régulée épigénétiquement par des facteurs viraux et cellulaires / High risk Human Papillomaviruses (HPV) are responsible for cervical cancer. HPV genome consists in a double-strand circular DNA harboring early "E" and late "L" genes and a Long Control Region (LCR). The E2 protcin binds to E2 Binding Sites (E2BS) present on the LCR and represses E6 and E7 transcription. The loss of E2 expression after HPV DNA integration induces an overexpression of E6 and E7 that thus favor p53 and pRb degradation. Since CpG dinucleotides are present in HPVl6 E2BS, we investigated whether E6 HPV16 expression was also submitted to epigenetic regulation. We developed a HRM PCR to study the methylation status of E2BS in precanccrous and canccrous lesions. We observed methylated CpG only in cancer samples. Otherwise, we proved that E2BS methylation prevented E2 binding and probably permitted E6 and E7 overexpression. Finally, we showed that the treatment ofHPV16 cervical cancer cell lines with a demethylating agent (SazadC) decreased the E6 expression. This regulation was independent of E2 and we proved that the up-regulation of miR-375, which targets E6/E7 transcripts, was involved in E6 repression after SazadC treatment. Taken as a whole, our data demonstrate that HPV 16 oncoprotein expression is regulated in an epigenetic manncr via viral and cellular factors.

O desenvolvimento de metodologias sobre integração de dados na área de biologia de sistemas é de grande importância devido ao aumento contínuo de dados resultantes de análises globais que são depositados em bancos de dados públicos. Poucas metodologias e ferramentas de bioinformática levam em consideração a diferenciação entre drivers e passengers, fundamental para a identificação de genes importantes para o desenvolvimento e progressão tumoral. Os perfis de expressão gênica têm possibilitado a identificação de assinaturas genéticas em uma grande variedade de tumores humanos. Além disso, as alterações epigenéticas, como a expressão de microRNAs (miRNA) e a metilação do DNA, também contribuem para o desenvolvimento de diversos tipos de doenças. Entretanto, a grande maioria destes estudos não mostra integração dos resultados obtidos pelas diferentes estratégias utilizadas, o que teria maior impacto na identificação de drivers moleculares. Neste estudo foi realizada a integração de quatro níveis de alterações em 31 amostras de câncer de pênis (CaPe): alteração do número de cópias do DNA, metilação de ilhas CpGs, expressão de miRNAs e expressão de transcritos codificadores. O conhecimento das alterações genéticas e epigenéticas relacionadas ao desenvolvimento de câncer de pênis é bastante limitado, devido principalmente a sua rara incidência. Uma parcela significativa dos casos de CaPe tem sido associada com a infecção pelo Papilomavírus Humano (HPV). A metodologia para integração de dados foi aplicada utilizando duas abordagens: (1) estudo das alterações em câncer de pênis (tumor e normal) independente da infecção pelo HPV e (2) estudo das alterações relacionadas à infecção pelo HPV. A análise foi dividida em duas etapas, com a seleção de genes alvos específicos da doença e inferência de módulos a partir desses alvos. Destacam-se na metodologia a seleção de genes candidatos a driver utilizando a atribuição de pesos para cada alteração seguindo critérios pré-determinados, e.g. se o gene estava presente em uma região rara, após classificação pelo DGV (Database of Genomic Variants) e a utilização desses genes como alvos para identificação das possíveis relações entre eles e os módulos. Também foi realizada a adaptação da metodologia de redes em módulos, com a inclusão de genes passengers e interação proteína-proteína (PPI) como um critério para seleção dos módulos. Essa análise se mostrou eficaz na identificação de módulos gênicos bem relacionados com os drivers, resultando na escolha de vias biológicas potencialmente responsáveis pelo desenvolvimento do tumor. Os genes identificados após a comparação entre amostras tumorais e normais (SOX17, TWIST1, CAV1, PPARG, FLI1 e TNFSF10) e no estudo entre amostras positivas e negativas para infecção pelo HPV (PCNA, SOX14 e RFC4) foram validados in situ por técnicas independentes. Para validação in silico das alterações encontradas na metodologia de integração de dados e para validação da metodologia de redes em módulos foram utilizadas 255 amostras de glioblastoma multiforme obtidas no banco de dados TCGA (The Cancer Genome Atlas). Foram identificadas vias biológicas importantes relacionadas ao processo tumoral, como regulação do crescimento celular (GO:0001558, p=0,0062), homeostase (GO:0048872, $p=0,0082) e regulação da transcrição (GO:0003700, p=0,00089). Também foi realizada uma meta-análise utilizando amostras do TCGA, que encontrou um perfil similar de expressão para os genes CAV1, DLC1,FLI1, MSX1, NRN1, PML, PPARG e SOX17 (T vs N) e PCNA e RFC4 (HPV+ vs HPV-). Para o nosso conhecimento, esse é o primeiro estudo em CaPe utilizando análise integrada de quatro níveis de alteração. Além disso, foram encontradas alterações não randômicas capazes de modificar transcritos específicos e contribuir para o conhecimento da patobiologia dos tumores de pênis. / Methodologies for data integration in systems biology area have great importance due to continuous increase of public data resulting from large-scale analysis, which are deposited in public databases. Few methodologies and bioinformatics tools take into consideration the differentiation between drivers and passengers, fundamental for the identification of important genes for tumor development and progression. The gene expression profiles have allowed the identification of genetic signatures in a wide variety of human tumors. In addition, epigenetic changes, such as the expression of microRNA (miRNA) and DNA methylation, also contribute to the development of a veriety of diseases. However, most of these studies did not show integration of results obtained by different strategies used, which would have increased impact to identify molecular drivers. This study provides a methodology for integration of four levels of changes in 31 samples of penile cancer (PeCa): copy number alterations, DNA methylation of CpG islands, miRNA expression and gene expression of coding transcripts. Knowledge about genetic and epigenetic changes related to the development of penile cancer is very limited, mainly due to its rare incidence. A significant portion of PeCa samples has been associated with infection by Human papillomavirus (HPV). The methodology for integrative data was applied using two approaches: (1) the study considering alterations in penile carcinoma (tumor and normal), independent of HPV infection and (2) the study considering alteration related to HPV infection in PeCa. In each study, the methodology was divided into two stages, with the selection of target genes and the inference of disease specific modules from these targets. It is highlighted in the methodology the selection of candidate genes using the driver assigning weights to each change following predetermined criteria, e.g if the gene was present in a rare region after classification using the DGV database (Database of Genomic Variants) and the use of these genes as seeds for identification of possible relationships between them and the modules. For this, another contribution of this study was the adaptation of module network methodology, with the inclusion of passengers genes and protein-protein interaction (PPI) as a criteria to select the modules. This analysis was effective in identifying gene modules and related drivers, resulting in the choice of biological pathway potentially responsible for the tumor development. The genes identified after comparing tumor and normal samples (SOX17, TWIST1, CAV1, PPARG, FLI1 and TNFSF10) and the genes identified in the study of positive and negative samples for HPV infection (PCNA, SOX14 and RFC4) were validated in situ by independent techniques. For in silico validation of the changes found in the integrative methodology and the modules network were used 255 samples of glioblastoma multiforme obtained at TCGA database (The Cancer Genome Atlas). Biological pathways have been identified related to the tumoral process, such as cell growth regulation (GO:0001558, p=0,0062), homeostasis (GO:0048872, p=0,0082) and transcription regulation (GO:0003700, p=0,00089). Also, a meta-analysis was performed using samples from TCGA, who found a similar expression profile for CAV1, DLC1, FLI1, MSX1, NRN1, PML, PPARG and SOX17 (T vs N) and PCNA and RFC4 genes (HPV + vs HPV-). To our knowledge, this is the first integrative analysis in PeCa using a four-level of gene alterations. In addition, it was found non-random alterations capable to modifying specific transcripts and contribute to the knowledge about the pathobiology of penile tumors.

Though vaccine uptake and public support have risen since the release of the first HPV vaccines, the United States has far lower initiation and completion rates for the HPV vaccine series in comparison to other vaccines indicated for youth. Disparities are even greater in the Appalachian regions. Understanding factors contributing to these discrepancies is vital to improving raise vaccine rates in Appalachia. A comprehensive literature search identified all articles pertaining to HPV vaccination in children and adolescents living in Appalachia. The final 15 articles were included in a systematic review of the topic. Findings: HPV disease and HPV vaccine-related knowledge and communication were low in Appalachian communities, and vaccine uptake was lower in all areas of Appalachia as compared to non-Appalachian U.S. Moreover, large variations in uptake existed among Appalachian subregions. Many variables appear to contribute to this variation, including vaccine acceptance for younger adolescents, local and press-driven critical reports of the vaccine, physician communication, and views of the family matriarchs. Targeting the Appalachian subregions, specific campaigns or intervention may be more impactful than viewing the region as a homogenous whole.

Background: Worldwide efforts have been made by some countries to offer HPV vaccination since its introduction in 2006. Population effectiveness of HPV vaccines is presently an active area of research. We review available evidence on the effectiveness of HPV vaccine uptake among female adolescents to prevent HPV infection. Methods: A comprehensive search of published and grey literature was conducted in several electronic databases using a pre-defined search strategy related to HPV prevalence following vaccination. The database searches were complemented by hand-searches of reference lists of eligible studies. Data were extracted onto a purpose-designed data extraction form, pooled in a meta-analysis and stratified by continent considering vaccine type, cross protective and (high/low) risk HPV types as subgroups. Results: Our search yielded 1680 studies, of which thirteen met with our inclusion criteria (8332 vaccinated women aged 12 to 34 years from across the world). The pooled HPV (comprising types 6, 11, 16 and 18) prevalence among vaccinated female adolescents was 7% (95% Confidence Interval (CI): 5% to 9%, 13 studies, n=8,332). The 13 studies were conducted across 3 continents: HPV prevalence for North America was 5% (95% CI: 3% to 7%, 9 studies, n=5781, age range =13 to 34); Europe, 14% (95% CI: 9% to 18%, 3 studies, n=2213, age range =13 to 29) and Australia 5% (95% CI: 3% to 8%, 1 study, n=5781, age range=13 to 34). Of the studies which reported the effect of vaccination on other non-vaccine HPV type prevalence (known as cross protective types) HPV (31, 33, 45, 51 &amp; 58), the overall pooled cross protective HPV prevalence was 9% (95% CI: 6% to 12%, 4 studies, n=3081 age range=13 to 29), by continent North America had 14% (95% CI: 12 to 17%, 1 study, n=753 age range=14 to 24), Europe 7% (95% CI: 6 to 8%, 2 studies, n=1990, age range=13 to 29) and Australia with 8% (95% CI: 5% to 11%, 1 study, n=338 age range=18 to 26). Conclusion: This study showed an HPV prevalence of 7% in women vaccinated against HPV types 6,11,16 and 18, which represents a substantial difference to the 22% HPV prevalence in non-vaccinated women. There was no statistically significant difference between HPV prevalence across the continents. There is however, still a dearth of information on vaccinated women and HPV prevalence, highlighting the need for further studies in this area. Strengths and limitation of this review • The review comprehensively searched multiple databases and bibliographies. We had no language restrictions. • We were stringent in the selection of studies as far as vaccination status was concerned. Studies considering HPV prevalence in unvaccinated women were excluded. • A variety of methods was utilised in collecting data across the studies. However, some of the study participants were not representative of the general population. Caution therefore, needs to be considered when using these results to make inferences or conclusions about prevalence of certain populations.

京都大学 / 0048 / 新制・課程博士 / 博士(生命科学) / 甲第17993号 / 生博第296号 / 新制||生||39(附属図書館) / 80837 / 京都大学大学院生命科学研究科統合生命科学専攻 / (主査)教授 藤田 尚志, 教授 杉田 昌彦, 教授 朝長 啓造 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM

No description available.

No description available.