Made available in DSpace on 2014-07-29T15:30:36Z (GMT). No. of bitstreams: 1 Dissertacao Andrea Alves Ribeiro.pdf: 1577076 bytes, checksum: 24bc7ace843786b03ff879da0df4f7aa (MD5) Previous issue date: 2009-12-15 / Human papillomavirus (HPV) is considered the central etiological agent involved in the genesis of cervical cancer. The HPV viruses are classified according to their biological niche, oncogenic potential and phylogenetic position. According to the criteria established by the International Committee on Taxonomy of Viruses (ICTV), the various groups of human papillomaviruses that infect the female genital tract are classified phylogenetically in the Alphapapillomavirus genus, including species classified among phylogenetic species 1 and species 15. The main high risk HPV are classified in species 9 (HPV 16, 31, 33, 35, 52, 58, 67), and in species 7 (18, 39, 45, 59, 56, 66, 68 and 70). HPV 16 is the most prevalent type irrespective of diagnosis, principally in more severe lesions. Coinfection with multiple-types HPV is a common finding of many molecular studies. Some HPV types might interact or act synergistically to induce progression. Few studies have investigated the interactions of viral genotypes or species in multiple-type HPV infections. Therefore, the objective of this study was to evaluate the effect of single or multiple-types HPV infections considering also the phylogenetic groups on the prevalence and severity of cervical intraepithelial neoplasia (CIN) among women undergoing colposcopy following a abnormal cervical smear. Methodology: In this analysis, 198 women attending at the colposcopic clinic, because of an abnormal cervical smear were included. Colposcopy was carried out in all cases and biopsies were done in 193 of 198 women included. All specimens were tested for 27 HPV genotypes by Roche s polymerase chain reaction reverse line blot assay. Results: The overall prevalence of HPV in women with an abnormal cervical smear was 86% (171/198). Of the total of HPV-positive women, 45% (77/171) were infected with HPV 16 as a single or multiple-type infections. HPV 31 and 35 were, respectively, the second and third most prevalent types. The prevalence of HPV 16 in high grade cervical intraepithelial neoplasia (CIN2/3) was 52% (40/76) and it was detected in 88.8% (8/9) in cases of invasive carcinoma. The prevalence of type 31 and 35 in high grade CIN was respectively 10.5% (8/76) and 6.6% (5/76). Single HPV infection for any type was significantly associated with neoplastic diagnosis. High grade neoplastic diagnosis (≥ CIN2) was significantly associated with HPV 16 in single or multiple infections. Also, there was significantly association between HPV 16 and others types of specie 9 and high grade neoplastic diagnosis, but no association was observed considering the HPV 16 and other of groups of species 7 or others types. Conclusion: These results indicated that the type 16 is the most important predictor of high grade cervical neoplasia. Multiple-type infections are predictors of high grade cervical neoplasia when the type 16 is present. / O Papilomavírus humano (HPV) é considerado o agente etiológico central envolvido na gênese do câncer cervical. O vírus HPV é classificado de acordo com seu nicho biológico, potencial oncogênico e classificação filogenética. De acordo com os critérios estabelecidos pelo Comitê Internacional de Taxonomia dos Vírus (ICTV), os diversos tipos de HPV que infectam o trato genital feminino são classificados filogeneticamente no gênero Alphapapillomavirus. Esta classificação inclui espécies filogenéticas classificadas entre a espécie 1 e a espécie 15, dentre as quais, as de maior interesse em relação ao potencial carcinogênico são a espécie 9 (HPV 16, 31, 33, 35, 52, 58, 67) e a espécie 7 (18, 39, 45, 56, 59, 66, 68, 70). O HPV 16 é tipo o mais predominante, independente do diagnóstico, presente principalmente nas lesões cervicais mais graves. A co-infecção com múltiplos tipos de HPV é um achado comum em muitos estudos moleculares, contudo, as interações dos genótipos virais ou espécies envolvidas nas infecções por múltiplos tipos de HPV têm sido pouco analisadas. Portanto, o objetivo deste estudo foi avaliar o efeito das infecções simples ou por múltiplos tipos de HPV, considerando também os grupos filogenéticos, sobre a prevalência e a gravidade das neoplasias cervicais. Metodologia: Este estudo de corte transversal incluiu 198 mulheres encaminhadas ao Ambulatório de Colposcopia da Santa Casa de Misericórdia de Goiânia por exame citopatológico anormal. Todas as mulheres foram esclarecidas quanto aos objetivos de estudo e assinaram o termo de consentimento livre e esclarecido. A colposcopia foi realizada em todos os casos e a biópsia em 193 das 198 mulheres incluídas. As amostras foram testadas para 27 genótipos de HPV, por reação em cadeia da polimerase (PCR); em seguida foi realizada a hibridização reversa em pontos da Roche Diagnósticos. Resultados: A prevalência de HPV em mulheres encaminhadas por exame citopatológico anormal foi de 86% (171/198). Do total de mulheres HPV-positivas, 45% (77/171) estavam infectadas por HPV 16 em infecções simples e múltiplas. Os tipos de HPV 31 e 35 foram respectivamente, o segundo e o terceiro mais prevalentes. A prevalência do HPV 16 foi de 52% (40/76) nas neoplasias intra-epiteliais cervicais de alto grau (NIC 2/3) e de 88,8% (8/9) nos casos de carcinomas invasivos. As prevalências dos tipos 31 e 35 em neoplasias intra-epiteliais cervicais de alto grau (NIC 2/3) foram de 10,5% (8/76) e 6,6% (5/76), respectivamente. A infecção simples por qualquer tipo de HPV foi significativamente associada com diagnósticos neoplásicos de alto grau (≥ NIC 2). Os diagnósticos neoplásicos de alto grau (≥ NIC 2) foram significativamente associados com o HPV 16 em infecções simples ou múltiplas, mesmo depois de ajustado pela positividade para DNA de HPV. Houve significativa associação entre o HPV 16 e outros tipos da espécie 9 e os diagnósticos neoplásicos de alto grau (≥ NIC 2), mas não foi observada associação, considerando o HPV 16 e outros tipos da espécies 7 ou outros tipos de HPV. Conclusão: Estes resultados indicam que o HPV 16 parece ser o mais importante preditor de diagnósticos neoplásicos de alto grau. As infecções múltiplas são preditoras das neoplasias cervicais de alto grau quando o HPV 16 está presente.

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This thesis focuses on gynecopathology. It consists of a collection of seven papers published in pathology journals with impact factor. Introduction section contains selection of examples showing scientific application of molecular genetic methods. Further on the aims of individual research projects are described. The first project comprises histomophologic study of skin endometriosis addressing "mullerian" differentiation. A case report of a rare tumor namely borderline papillary serous tumor of the fimbriated end of the fallopian tube follows with molecular genetic analysis of KRAS, BRAF and p53 gene mutation status. Prospective longitudinal study on high grade squamous dysplasia (HSIL) of the cervix in HPV vaccinated women, so called DAV (dysplasia after vaccination), aims to elucidate pathogenesis of this phenomenon. Two other studies focus on incidence of fumarate hydratase deficient leiomyomas of the uterus and hereditary leiomyomatosis and renal cell carcinoma syndrome (HLRCC). The aim of those studies is to improve our diagnostic capability and increase detection rate of the patients with HLRCC syndrome. Finally a new subtype of HSIL namely bizarre cell dysplasia is described in two separate studies. Conclusion remarks contemplate the role of molecular genetics in surgical pathology.

The MHC class I status of tumour cells during immunotherapy is often underestimated. It represents one of important tumour escape mechanisms and thus can contribute to the failure of most of the cancer clinical trials that are usually based on the induction of cytotoxic T cell responses. Epigenetic changes in the promoters of genes involved in the MHC class I Ag presentation can result in decreased expression of the cell surface MHC molecules on tumour cells. Thus, epigenetic modifiers can restore an expression of the MHC class I molecules and make tumours visible to the CD8+ effector cells. Besides the epigenetic changes on the tumour cells, epigenetic modulators affect cells of the immune system such as dendritic cells (DC). Tumour cells can escape from the immune response not only by changes in the cancer cells, but also by influencing, expanding and/or activating immunoregulatory cell populations, such as regulatory T cells (Treg). This thesis focuses on the potential of the DC-based vaccines against HPV-16-associated tumours with a different MHC class I expression, on the combination of cancer immunotherapy with the treatment using epigenetic modifiers, with special attention paid to their effects on DC, and, finally, on the impacts of the anti-CD25 antibody (used for Treg elimination) on Treg and NKT...