Submitted by Maria Creuza Silva (mariakreuza@yahoo.com.br) on 2013-10-03T19:10:15Z No. of bitstreams: 1 Diss MP. Ieda Carvalho 2013.pdf: 743354 bytes, checksum: 86a38cc8c18f66fee690c10a1336c161 (MD5) / Approved for entry into archive by Maria Creuza Silva(mariakreuza@yahoo.com.br) on 2013-10-03T19:10:31Z (GMT) No. of bitstreams: 1 Diss MP. Ieda Carvalho 2013.pdf: 743354 bytes, checksum: 86a38cc8c18f66fee690c10a1336c161 (MD5) / Made available in DSpace on 2013-10-03T19:10:31Z (GMT). No. of bitstreams: 1 Diss MP. Ieda Carvalho 2013.pdf: 743354 bytes, checksum: 86a38cc8c18f66fee690c10a1336c161 (MD5) Previous issue date: 2013 / O câncer de colo de útero (CCU) é um importante problema de saúde em todo o mundo. O HPV é o principal fator associado a esta doença. Para a prevenção primária da infecção foi desenvolvido a vacina quadrivalente que protege contra os tipos HPV 16, 18, 11 e 6. Objetivo: este estudo analisou a relação custo-efetividade e custo-utilidade da adição da vacina contra HPV para Sistema Único de Saúde brasileiro, em comparação ao rastreamento pelo exame citopatológico, programa existente. Método: foi adaptado um modelo de Markov da história natural da infecção por HPV para estimar custo e qualidade de vida para uma coorte hipotética de meninas de 10 anos de idade acompanhadas por 70 anos. Duas estratégias foram comparadas: vacinar e rastrear (estratégia alternativa) em relação ao rastreamento (estratégia base). No modelo a cobertura para o rastreamento foi de 77,1% e para a vacina 95%. A taxa de desconto aplicada para calcular custos e efeitos futuros de saúde foi de 5%. A análise de sensibilidade foi realizada para avaliar as incertezas quanto à cobertura vacinal, a sensibilidade do exame citopatológico e o valor da vacinação. Resultados: o modelo simulou que a adição da vacina poderia reduzir em 76,18% o risco de morrer por CCU, evitando 12.072 óbitos por esta doença a um custo médio de US$ 633.559,32/óbitos evitados, com um ganho de 1.389.478 anos de vida ajustado por qualidade. Quanto aos anos de vida ganhos ajustado por qualidade (AVAQ), a estratégia base (rastreamento) representaria um custo médio de US$ 11,62/AVAQ enquanto a estratégia teste (vacina e rastreamento) teria um custo médio de 241,66 US$/ AVAQ. A razão de custo-efetividade incremental (RCEI) desta estimativa foi de US$ 5.504,46/AVAQ. Conclusão: Esta análise, apesar das limitações metodológicas, demonstra que a incorporação, pelo SUS, da vacina quadrivalente ao programa de rastreamento pode ser uma alternativa custo-efetiva para reduzir a mortalidade por CCU. / Salvador

The human immunodeficiency virus is presently one of the most significant global health issues to date, with a disease burden that encumbers developing and developed nations alike. Although current antiretroviral therapy can help patients maintain undetectable levels of the virus throughout their bodies, once the treatment is ceased, the virus will rebound and disease progression continues. Thus, modalities to; 1- stop HIV transmission and spread, or 2- eradicate the virus once it is acquired are both urgently needed. In this project, we seek to evaluate and understand the impact of a candidate vaccine therapy that targets the HIV protease cleavage sites (PCS) on viral fitness. Vaccination with this modality in a monkey model induces mutations at virus regions that are intolerant to change, presumably affecting the "fitness" of viral strains recovered from vaccines. Preliminary results of the study show that in the vaccine group (n=11), a disruption to one or more of the HIV protease cleavage sites results in improved maintenance of CD4+ T cells compared to unvaccinated controls (n=5). Furthermore, a correlation between the percentage of PCS mutations and reductions in viral load were seen. Our data indicate that the most common sites of mutation occur at two cleavage regions PCS2 and PCS12. We used site directed mutagenesis to introduce multiple PCS mutations into infectious clones of SIV. Our ongoing studies are evaluating the viral fitness of the SIV mutants in a cell lines and PBMC using competitive viral fitness assays. The data from these studies will help inform in the areas of vaccine and therapy development for HIV-1.

Cervical cancer is one of the most common forms of cancer in the world, and human papillomavirus (HPV) is a cause of the vast majority of these patient cases. With many HPV types being oncogenic in nature, HPV as a whole is responsible for over 5% of all cancers worldwide and 15% of cancer in women in developing countries. HPV is a sexually transmitted infection that is spread through contact with infected genital skin, mucosa, or bodily fluids from a partner with acute or subclinical viral infection. While less frequent, various strains of the virus are also responsible for anal and vaginal warts, anal cancer, and cancer of the vulva and penis – these account for approximately 50,000 cases per year worldwide. Data also suggest a potential implication of HPV in oropharyngeal cancers, especially among younger adults. Various behavioral and prophylactic approaches are recommended for the prevention of HPV infection and cancer. For example, there is evidence that behavioral change can be effective, such as condom use and limitation on the number of sexual partners. Besides this, in recent years we have seen the development of various prophylactic or therapeutic vaccines that are highly effective in the prevention of HPV pathogenesis. Despite this, barriers to treatment and prevention exist, making HPV a continuing threat to individuals most at-risk across the globe. Thus, this study reviewed a large collection of current HPV and related cancer literature to understand the process of infection and pathogenesis in various human sites as well as potential barriers to prevention and treatment that may be perpetuating the survival of the virus across the world. Analyzing current and past research on such barriers, this paper delves into important variables that can affect early detection and treatment of HPV, and also explores a novel and promising therapy currently in development that could be valuable in overcoming many of these issues.

A thesis by Marc Ashley Bayliss entitled ‘Virus-like particles as a novel platform for delivery of protective Burkholderia antigens’ and submitted to the University of Exeter for the degree of Doctor of Philosophy. There is currently no licensed vaccine available for the global tropical pathogen Burkholderia pseudomallei which is the causative agent of melioidosis and a potential bio-threat agent. The capsule polysaccharide (CPS) expressed by B. pseudomallei has been shown to offer some protection against bacterial challenge. Polysaccharide immunogenicity can be enhanced by conjugation to a carrier protein and several licensed vaccines utilise this technology. Virus-like particles (VLPs) are non-infectious, non-replicating, viral proteins that self-assemble into viral structures and are in several licensed vaccines as primary antigens. VLPs are also effective delivery platforms for foreign antigens by genetic insertion or chemical conjugation. iQur, a collaborator on this project, has developed Tandem CoreTM that consists of two genetically linked hepatitis B core proteins that allow insertion of large proteins into each core whilst remaining assembly competent. The aim of this thesis was to assess the protective efficacy of Tandem CoreTM VLPs chemically conjugated to CPS and Tandem CoreTM Burkholderia protein fusion constructs. This involved three objectives; reduce the cost of CPS extraction; identify immunogenic Burkholderia proteins; and test candidate vaccine efficacy in an animal model of acute melioidosis against B. pseudomallei challenge. To reduce the cost of extraction, CPS was purified from B. thailandensis strain E555 and bacterial culture CPS concentration optimised which first required development of a quantitative ELISA. Immunogenic Burkholderia proteins were identified from the literature but Tandem CoreTM fusion constructs containing these proteins were not assembly competent. The Burkholderia proteins were added as co-antigens to the VLP CPS conjugate vaccine but did not improve efficacy. Tandem CoreTM VLPs conjugated to CPS were protective against B. pseudomallei challenge and were compared to CPS conjugated to Crm197: a commercially available carrier protein used in several licensed vaccines. At lower challenge doses, survival was greater in mice vaccinated with the VLP-CPS conjugate although at higher doses, Crm197-CPS efficacy was greater.

Ticks acquire nutrients only by a parasitic nature of feeding on animals, including humans. During this process, a wide array of pathogens is transmitted. Ticks of the Ixodidae family receive exactly one blood meal in each active developmental. Knowing the trophic dependence of tick metabolism on the host blood meal components may enable discovering processes essential for the tick physiology and development. Exploiting a membrane system of tick feeding and whole blood fractionation, we have revealed that adult ticks need to acquire host haemoglobin-derived haem so that they can produce viable larvae, and reproduce. Haem is not further catabolised in ticks, and iron is thus acquired via independent route with the host serum transferrin as a source molecule. Using RNA-seq, we compared transcriptome compositions between guts of blood- and serum-fed ticks. We identified fifteen gut transcripts that change their levels with respect to the presence/absence of dietary red blood cells. Glutathione S-transferase, one of the identified encoded molecules, shows a clear haeminresponsive expression at both transcript and protein levels. Its apparent haem-binding properties suggest that this protein is directly involved in haem homeostasis maintenance within the tick gut. The ultimate goal of such research is to identify and verify targets that, when blocked, would render the acquisition and/or distribution system of haem in ticks nonfunctional. This would represent a novel way of anti-tick interventions in veterinary and human medicine.

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2013-10-14T18:17:57Z No. of bitstreams: 1 Lais da Silva Pereira. Avaliação das histonas...2013.pdf: 2452594 bytes, checksum: 808d7633f6d2684df54ea8b640e282c0 (MD5) / Made available in DSpace on 2013-10-14T18:17:57Z (GMT). No. of bitstreams: 1 Lais da Silva Pereira. Avaliação das histonas...2013.pdf: 2452594 bytes, checksum: 808d7633f6d2684df54ea8b640e282c0 (MD5) Previous issue date: 2013 / Universidade Federal da Bahia. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / A leishmaniose visceral é uma doença infecciosa grave, causada por protozoários intracelulares obrigatórios do gênero Leishmania. Vários antígenos de Leishmania têm sido avaliados como candidatos vacinais, destacando-se as proteínas de histonas (HIS), antígenos altamente conservados. A exposição de HIS pela Leishmania induz uma resposta imune potente no hospedeiro vertebrado. Desse modo, neste estudo, avaliamos, em hamsters, a capacidade imunoprotetora dos antígenos de histonas contra a infecção por Leishmania infantum. Os animais foram vacinados com estratégia homóloga, utilizando-se plasmídeos de DNA que codificam para HIS (pcDNA3LiH2A-H3, pcDNA3LiH2B-H4) ou heteróloga (DNA/proteínas HIS) mais 1nM de CpG. Quinze dias após a última imunização, os animais foram desafiados pela via intradérmica com 105 Leishmania infantum metacíclicas mais 0,5 par de glândula salivar de Lutzomya longipalpis. Após a última imunização e durante a infecção, realizaram-se dosagens de citocinas por PCR em tempo real (linfonodo e baço), sorologia por ELISA (soro), carga parasitária por diluição limitante e análise histopatológica de tecidos (linfonodo, baço e fígado). Detectou-se produção de anticorpos IgG anti HIS nos grupos imunizados com a estratégia homóloga e heteróloga, quando comparados aos hamsters não imunizados. As imunizações homóloga e heteróloga diferiram na razão IFN-γ/IL-10 no linfonodo em relação ao grupo controle. Não houve diferença significativa na expressão dessas citocinas no baço após a imunização, entretanto, cinco meses após o desafio o grupo homólogo apresentou um aumento na produção de IL-10 nesse órgão. Na análise histopatológica do baço, verificou-se formação de mais folículos com centro germinativo, evidentes nos animais imunizados independentemente do grupo analisado. Observou-se, também, leucocitose intrasinusoidal e periportal no fígado, e folículos reativos no linfonodo. Nenhuma das estratégias de imunizações com antígenos de histonas acarretou em diminuição da carga parasitária no linfonodo, baço e fígado. As estratégias de imunização homóloga e heteróloga, com antígenos de histonas nucleossomais, não foram capazes de proteger contra infecção por L. infantum no modelo do hamster. / Visceral leishmaniasis is a serious infectious disease caused by obligatory intracellular protozoan of the Leishmania genus. Many antigens from Leishmania have been evaluated as vaccine candidates mainly the histones (HIS) that are highly conserved. Exposure to HIS from Leishmania elicits a strong host immune response in the vertebrate host. Threfore, in this study, we evaluated the immunoprotective ability of HIS antigens against infection by Leishmania infantum in hamsters. The animals were immunized with homologous strategy using plasmid of the DNA coding for HIS (pcDNA3LiH2A-H3, pcDNA3LiH2B-H4) or heterologous strategy using DNA and recombinant protein plus CpG. Fifteen days after last immunization, the animals were challenged by the intradermal route with 105Leishmania infantum metacyclics plus 0,5 pair of Lutzomya longipalpis salivary gland. After the last immunization and in the course of the infection, we determined cytokine production by Real time PCR (lymph node and spleen), serology by ELISA (sera), parasite burden was estimated by limiting dilution assay and histopathological analysis (lymph node, spleen and liver). Production of antibody anti HIS IgG was detected in the immunized groups treated with the homologous and heterologous strategies when compared to non-immunized hamsters. Homologous and heterologous immunizations altered the ratio of IFN-γ/IL-10 in the lymph node compared to the control group. There is not a significant difference in the expression of this cytokine in the spleen after the immunization, however, five months after the challenge the homologous group presented a higher expression of IL-10 in this organ. The histopathological analysis of the spleen showed the formation of high number of follicles with a germinative center in the immunized animals independently of the immunization employed. Intrasinusoidal and periportal leukocytosis were observed in the liver and reactive follicles in the lymph node. Both immunization strategies with histone antigen did not result in reduction of parasite burden in the lymph node, spleen and liver. Immunization with homologous and heterologous strategies using histone antigen were not able to confer protection in the hamster model against L. infantum infection.

In regions endemic for malaria, natural exposure results in an acquired immunity which protects individuals from severe disease. However, no vaccine against the blood-stage of malaria, against which naturally-acquired immunity is targeted, currently exists that is capable of emulating, or out-performing, natural protection. To rationally direct the next generation of blood-stage malaria vaccine development, a greater understanding of the immunological mechanisms involved in clinical protection is required. To date, the assessment of naturally-acquired and vaccine-induced immunity to the blood-stage of malaria has suffered from a paucity of in vitro immunological assays that are both robust and reproducible, whilst allowing for assessment of anti-parasitic activity induced by antibodies, either alone, or in conjunction with immune cells. Thus this Thesis describes the development of the antibody-dependent respiratory burst (ADRB) assay, for assessment of blood-stage immunity against Plasmodium falciparum, as well as the Duffy antigen receptor for chemokines (DARC) – Duffy binding protein (DBP) binding inhibition assay, for assessing antibody mediated immunity to P. vivax. A reproducible and standardised assay of ADRB activity was developed here and applied to studies of immunity in both mice and humans. ADRB activity, which assesses antibodies' ability to activate oxidative burst in neutrophils via Fc receptor (FcR)-dependent pathways, was shown to associate with clinical protection in a cohort from Mali where FcR-independent immunological assays, such as the assay of growth inhibition activity, did not. This work thus elucidates the importance of FcR-dependent immunity to P. falciparum malaria and establishes the ADRB assay as a useful tool for future vaccine development. In addition, the DARC-DBP binding inhibition assay was established and utilised to assess inhibitory activity of antibodies induced in the first Phase I clinical trial of this antigen. Results identify the need for significant improvements in vaccine design, and show the utility of the assay as a tool for assessing future blood-stage vaccine development efforts against this neglected parasite.

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2012-08-01T18:43:17Z No. of bitstreams: 1 Naiara Carvalho Teixeira Imunização de camundongos com fragmentos....pdf: 902012 bytes, checksum: 1ec320ac38a55ad96f41e4c20bcf883d (MD5) / Made available in DSpace on 2012-08-01T18:43:17Z (GMT). No. of bitstreams: 1 Naiara Carvalho Teixeira Imunização de camundongos com fragmentos....pdf: 902012 bytes, checksum: 1ec320ac38a55ad96f41e4c20bcf883d (MD5) Previous issue date: 2011 / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, Brasil / Leishmaniose visceral zoonótica (LV) é uma doença que afeta homens e cães e é causada por protozoários das espécies Leishmania infantum e Leishmania chagasi. O cão doméstico é o principal reservatório do agente causal. Uma vacina efetiva contra a LV canina poderá contribuir para o controle da infecção e/ou doença humana e canina. A resposta imune protetora contra a LV canina é do tipo celular (Th1). Em nosso laboratório, dentre um grupo de cinco antígenos recombinantes selecionados de uma biblioteca de cDNA de L. chagasi, usando-se uma mistura de soro de cães naturalmente infectados e que exibiam resposta imune humoral e celular específica, um fragmento da extremidade carboxila de uma cinesina com cauda de 6 histidinas (rLci2B-NH6), para permitir a purificação, foi escolhido para a avaliação do seu potencial como candidato a componente de uma vacina contra LV canina. No presente trabalho foi avaliada a imunização de camundongos BALB/c com rLci2BNH6/ saponina em associação com os plasmídeos pcDNA3.1-scmu-IL-12 e/ou pcDNA3.1- mu-IL-2. Os animais imunizados com rLci2B-NH6/saponina em associação com a IL-12 e/ou IL-2 produziram anticorpos IgG, IgG2a e IgG1 específicos. Esplenócitos desses animais exibiram proliferação, mas não produziram IFN-γ ou IL-5 após estimulação in vitro com rLci2B-NH6. Esses resultados sugerem que a adição de pcDNA3.1-mu-IL-2 no protocolo de imunização não é capaz de promover uma resposta imune predominantemente Th1. Também foi avaliada a imunização de camundongos com proteína recombinante quimérica (rLci2-NTCT- NH6) formada pelos domínios não repetitivos presentes nas extremidades amina e carboxila codificadas pelo gene da cinesina. Camundongos BALB/c imunizados com rLci2- NT-CT-NH6 ou uma proteína recombinante químerica controle, contendo 5 domínios repetitivos presentes em rLci2B-NH6 entre os domínios não repetitivos de rLci2-NT-CTNH6, apresentaram produção de IgG, IgG2a e IgG1. Esplenócitos desses animais exibiram proliferação, mas não produziram IFN-γ e apenas esplenócitos dos camundongos imunizados com rLci2-NT-5R-CT-NH6 sintetizaram IL-5, após estimulação in vitro com as respectivas proteínas. Contudo, durante a purificação de rLci2-NT-CT-NH6 e rLci2-NT-5R-CT-NH6 ocorreu intensa proteólise que pode ter resultado na destruição de epítopos capazes de induzir resposta Th1. Previamente, em nosso laboratório, foi observada a morte de alguns camundongos injetados com 100 μg saponina/dose, por isso, doses menores de saponina (50 ou 25 μg) foram avaliadas como adjuvante na indução de resposta imune a rLci2B-NH6. Os resultados da mensuração dos anticorpos foram semelhantes nos três grupos. Curiosamente, somente esplenócitos do grupo injetado com 25 μg/dose exibiram proliferação específica. O grupo injetado com 100 μg/dose produziu IFN-γ e IL-5 e o grupo injetado com 50 μg/dose sintetizou IFN-γ, mas não produziu IL-5, após estimulação in vitro com rLci2B-NH6. / Zoonotic visceral leishmaniasis (VL) is a disease that affects humans and dogs and is caused by protozoan species Leishmania infantum and Leishmania chagasi. There is evidence to suggest that the domestic dog is the main reservoir of the causative agent. An effective vaccine against canine VL may contribute to infection control and/or human and canine disease. The protective immune response against canine VL is cellular (Th1 type). In our laboratory, among a group of five recombinant antigens selected from a cDNA library of L. chagasi, using a mixture of sera from naturally infected dogs that showed humoral and cellular specific responses, one fragment of the carboxyl terminus of a kinesin with a 6 His- Tag (rLci2B-NH6) was chosen to evaluate its potential as a candidate component of a vaccine against canine VL. In the present study, we evaluated the immune response of BALB/c immunized with rLci2B-NH6/saponin in combination with pcDNA3.1-scmu-IL-12 and pcDNA3.1-mu-IL-2. The animals immunized with rLci2B-NH6 associated IL-12 and/or IL-2 produced IgG, IgG2a and IgG1 reactive to rLci2B-NH6. Splenocytes of these animals exhibited proliferation but failed to produce IFN-γ and IL-5 after in vitro stimulation with rLci2B-NH6. These results suggest that adding IL-2 was not enough to induce a predominantly Th1 immune response. We also analyzed the immunization of mice with recombinant chimeric (rLci2-NT-CT-NH6) formed by non-repetitive domains present in the amine and carboxyl ends encoded by the gene for kinesin. BALB/c mice immunized with rLci2-NT-CT-NH6 or a control chimeric recombinant protein containing 5 repeated domains present in rLci2B-NH6 between the domains of non-repetitive rLci2-NT-CT-NH6 showed production of IgG, IgG2a and IgG1. Splenocytes of these animals exhibited proliferation, but failed to produce IFN-γ and only splenocytes from mice immunized with rLci2-NT-5R-CTNH6 synthesized IL-5 after in vitro stimulation with the respective proteins. However, during the purification rLci2-NT-CT-NH6 and rLci2-NT-5R-CT-NH6 occurred intense proteolysis that may have resulted in the destruction of epitopes capable of inducing Th1. Previously, in our laboratory, it was observed the death of some mice injected with 100 μg saponin/dose, therefore, lower doses of saponin (50 or 25 μg) were evaluated as an adjuvant to induce an immune response to rLci2B-NH6. The results of measuring the antibodies were similar in the three groups. Interestingly, only splenocytes from the group injected with 25 μg/dose exhibited specific proliferation. The group injected with 100 μg/dose produced IFN-γ and IL- 5 and the group injected with 50 μg /dose synthesized IFN-γ, but did not produce IL-5 after in vitro stimulation with rLci2B-NH6.

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2016-05-11T14:07:03Z No. of bitstreams: 1 Jailton_Azevedo Silva Junior. Impacto da vacina...2015.pdf: 2256825 bytes, checksum: 6da623ba97a2d68604f3b4a098585b07 (MD5) / Approved for entry into archive by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2016-05-11T14:07:15Z (GMT) No. of bitstreams: 1 Jailton_Azevedo Silva Junior. Impacto da vacina...2015.pdf: 2256825 bytes, checksum: 6da623ba97a2d68604f3b4a098585b07 (MD5) / Made available in DSpace on 2016-05-11T14:07:15Z (GMT). No. of bitstreams: 1 Jailton_Azevedo Silva Junior. Impacto da vacina...2015.pdf: 2256825 bytes, checksum: 6da623ba97a2d68604f3b4a098585b07 (MD5) Previous issue date: 2015 / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / INTRODUÇÃO: Em 2010, a vacina conjugada 10-valente (PCV10) foi incorporada ao programa nacional de imunizações (PNI) brasileiro. Este imunobiológico confere imunização contra os dez principais tipos capsulares de Streptococcus pneumoniae, patógeno responsável por diversas manifestações clínicas e com elevada contribuição nas taxas de incidência e mortalidade por meningite, que é a condição clínica mais grave. OBJETIVO: O presente estudo teve como objetivo avaliar o impacto da PCV10 na epidemiologia da meningite pneumocócica na região metropolitana de Salvador (RMS) Bahia, comparando o período anterior (2008-2010) e posterior (2011-2013) a sua utilização, bem como realizar uma caracterização molecular minuciosa a partir de uma série histórica (1996-2012) entre os isolados resistentes a penicilina (PNSSP com CIM≥ 0,125 μg/mL) e para os sorotipos não-vacinais (2008-2012). MATERIAL E MÉTODOS: Foram incluídos todos casos de meningite pneumocócica confirmados laboratorialmente no período entre 1996 a 2013. Taxas de incidência para a Salvador e RMS foram calculadas com base nos dados populacionais do IBGE/2010. A determinação do tipo capsular foi realizada através da técnica de Multiplex-PCR e/ou reação de Quellung. A sensibilidade a nove antimicrobianos foi testada através das técnicas disco-difusão, microdiluição e E-test. Para caracterizar o perfil molecular foram aplicadas as técnicas de genotipagem de PFGE e MLST. RESULTADOS: Um total de 939 casos de meningite pneumocócica foram identificados no período de 1996- 2013, sendo que 70 casos ocorrem entre 2011 a 2013 (período pós-vacinal). A incidência de meningite pneumocócica em todas as faixas etárias na RMS reduziu de 0,70 casos/100.000 habitantes para 0,59 casos/100.000 habitantes considerando o período de três anos antes e após a vacinação com PCV10 [p< 0,05; RR IC 95%: 1,46 (1,03-2,05)]. Esta redução foi significativa na faixa etária de 0-2 anos e nos casos por sorotipos relacionados à PCV10. Não houve aumento significativo de casos por sorotipos não vacinais nesta casuística, apesar do surgimento de casos por sorotipos não-vacinais não detectados anteriormente na série histórica de MP (10F, 21, 22F, 15A e 24F). Os isolados resistentes à penicilina analisados na série histórica se restringiram a 13 sorotipos, entre os quais: 14 (45,1 %; 78/173), 23F (19,1%; 33/173), 6B (14,4 %; 25/173), 19F (9,2 %; 16/173) e 19A (5,2 %; 9/173). 94% dos casos nãosusceptíveis à penicilina (PNSSP) foram de sorotipos vacinais. Os grupos clonais caracterizados pelo PFGE/MLST predominantes ao longo dos anos foram representados pelo sorotipo 14, denominado grupo A/ST 66 [35,3 % (61/173)] e grupo GK/ST 156 [4.6 % (8/173)], este último associado com níveis elevados de resistência a penicilina e ceftriaxona. Não foram detectados grupos clonais emergentes associados a tipos capsulares não-vacinais. CONCLUSÕES: Estes achados sugerem que a introdução da PCV10 modificou a epidemiologia da meningite pneumocócica na população estudada. / INTRODUCTION: In 2010, the 10-valent pneumococcal conjugate vaccine (PCV10) was introduced into the Brazilian national immunization program (NIP). This immunobiological provides immunization against the main ten capsular types of Streptococcus pneumoniae, the pathogen responsible for different clinical manifestations and high contribution in the incidence and mortality from meningitis, which is the most severe clinical condition. OBJECTIVE: This study aimed to evaluate the impact of PCV10 in the epidemiology of pneumococcal meningitis in the metropolitan area of Salvador (RMS) Bahia, comparing the previous (2008-2010) and after (2011-2013) periods its use, as well as conduct a thorough molecular characterization from a historical series (1996-2012) among isolates resistant to penicillin (PNSSP with CIM≥ 0.125 g / ml) and nonvaccine serotypes (2008-2012). MATERIAL AND METHODS: We included all cases of pneumococcal meningitis laboratory confirmed for the period 1996 to 2013. Incidence rates for Salvador and RMS were calculated based on population data from IBGE/2010. The capsular type determination was performed by multiplex PCR and/or Quellung reaction. Isolates Nine antibiotics were tested by disk-diffusion test, broth micro-dilution and E-test. To characterize the molecular profiling techniques were applied genotyping PFGE and MLST. RESULTS: A total of 939 cases of pneumococcal meningitis were identified during 1996-2013 period, with 70 cases occurring between 2011-2013 (post-vaccination period). The incidence of pneumococcal meningitis in all age groups in the RMS decreased from 0.70 cases / 100,000 inhabitants to 0.59 cases / 100,000 inhabitants considering the three-year period before and after vaccination with PCV10 [p <0.05; RR 95% CI: 1.46 (1.03 to 2.05)]. This reduction was significant in the age group 0-2 years and in cases by serotypes related to PCV10. There was no significant increase in cases by serotypes not vaccine in this series, despite the emergence of cases by serotypes not-vaccine previously undetected in the historical series of MP (10F, 21, 22F, 15A and 24F). The penicillin resistant isolates analyzed the historical series were restricted to 13 serotypes, including: 14 (45.1%; 78/173), 23F (19.1%; 33/173), 6B (14.4%; 25/173), 19F (9.2%, 16/173) and 19A (5.2%, 9/173). 94% of nonsusceptible to penicillin cases (PNSSP) were vaccine serotypes. Clonal groups characterized by PFGE / MLST predominant over the years have been represented by serotype 14, group called A / ST 66 [35.3% (61/173)] and Group GK / TS 156 [4.6% (8/173) ], the latter associated with elevated levels of penicillin and ceftriaxone resistance. Not were detected emerging clonal groups associated with capsular types non-vaccination. CONCLUSIONS: These findings suggest that the introduction of PCV10 changed the epidemiology of pneumococcal meningitis in the population studied.

Hepatitis C virus (HCV) infection is a major global health burden affecting 1-2% of the world’s population. The majority of infected individuals will develop chronic infection and are at risk of cirrhosis and hepatocellular carcinoma. There is currently no preventative vaccine available for HCV. In the developed world, the highest HCV incidence and prevalence rate is amongst intravenous drug users (IDU). The duration, frequency of IDU, and sharing of drug injecting equipment contribute to particularly high rates of HCV infection in this population. Individuals at high risk of recurrent exposure to HCV infection from long term IDU have been recruited in Plymouth, UK, from 2003 onwards and if they remain negative for HCV infection are termed exposed uninfected (EU). Understanding the factors that prevent HCV infection in this cohort could give valuable insight into the mechanisms of natural resistance to HCV infection. The EU cohort was previously shown to have characteristics attributable to the activation of both the adaptive and the innate arms of the immune system with no known dominant, immune or non- immune, mechanism of HCV protection. The aim of this thesis was to attempt and identify this mechanism and for that purpose a comparative transcriptional profile study was initially performed between 3 groups: EU, individuals who spontaneously cleared HCV infection (SR) and patients with chronic HCV infection (CHCV). Of the differentially regulated genes, the association with resistance to HCV was strongest for Interleukin-27 (IL-27) which was significantly upregulated in EU compared to the 2 other groups and C X C motif chemokine 7 (CXCL7) which was significantly upregulated in EU relative to the CHCV group. The CD28 mediated T-helper cell signalling pathway was significantly upregulated in SR relative to the 2 other groups. We attempted to corroborate the above findings and we demonstrated that IL-27 is overexpressed in EU, compared to SR and CHCV. The possible role of IL-27 in natural protection from HCV infection remains to be elucidated and requires further study.