Chlamydia trachomatis (C. trachomatis) is the most prevalent sexually transmitted bacteria with devastating reproductive consequences that lead to tubal factor infertility (TFI). Recent studies have implicated apoptosis – associated speck – like protein containing a caspase recruitment domain (ASC) as an adaptor of inflammasomes that stimulate IL – 1β and IL – 18 secretion, pro – inflammatory cytokines with critical functions in host defense against a variety of pathogens. Therefore, for the first time, we are reporting the use of ASC-/- mice in a mouse model of Chlamydia infection that might provide some information on the role of inflammasomes in the pathogenesis of Chlamydia infection. In this study, wild type (WT) and ASC-/- mice were infected with Chlamydia. Infectivity, pathology of the upper genital tract (UGT), and, fertility were evaluated. In addition, expression of ASC – dependent inflammasomes and the activation of immune cells within the genital tract (GT) were studied. Results showed that Chlamydia infectivity in ASC-/- mice was significantly higher (p-/- mice which, when compared to infected WT mice, was exhibited by decrease in average number of pups and percent pregnancy. There was also severe UGT damage in ASC-/- mice compared to WT mice, correlating with the higher number of hydrosalpinx observed on the UGT of Chlamydia infected ASC-/- mice. Furthermore, IL – 1β and IL – 18 production as well as immune cell activation were down regulated in the GT of Chlamydia infected ASC-/- mice. This finding indicates that in absence of ASC, host innate and adaptive immunity is impaired. Results imply that ASC plays a protective role in the mucosal immunity against GT Chlamydia infection.

To detect viral infections and tumors, CD8 T lymphocytes monitor cells for the presence of antigenic peptides bound to MHC class I molecules. The majority of MHC class I-presented peptides are generated from the cleavage of cellular and viral proteins by the ubiquitin-proteasome pathway. Many of the oligopeptides produced by this process are too long to stably bind to MHC class I molecules and require further trimming for presentation. Cytosolic aminopeptidases such as leucine aminopeptidase (LAP), which is IFN-inducible, Bleomycin Hydrolase (BH), and puromycin-sensitive aminopeptidase (PSA) can trim precursor peptides to mature epitopes and have been thought to play an important role in antigen presentation. To examine the role of these aminopeptidases in generating MHC class I peptides in vivo, we generated mice deficient in LAP or PSA, as well as cell lines deficient in LAP, PSA, or BH. LAP mutant mice and cells are viable and grow normally, whereas PSA mutant mice are smaller than their wild-type and heterozygote littermates, are subfertile as adults, and are subviable as embryos. The trimming of peptides in LAP-deficient cells is not reduced under basal conditions or after stimulation with IFN. Similarly, there is no reduction in presentation of peptides from precursor or full length antigen constructs or in the overall supply of peptides from cellular proteins to MHC class I molecules, even after stimulation with IFN. After viral infection, LAP-deficient mice generate normal CTL responses to seven epitopes from three different viruses. Similarly, PSA deficient mice and BH deficient mice generate normal CTL responses to viral epitopes. These data demonstrate that LAP, BH, and PSA are not essential enzymes for generating most MHC class I-presented peptides and reveal redundancy in the function of cellular aminopeptidases in most cell types.

The Tec family tyrosine kinases Itk, Tec and Rlk are expressed in T cells. Previous studies have established that these kinases are critical for TCR signaling, leading to the activation of PLCγ1. To further understand the functions of Tec kinases in T cell activation, we took three different approaches. First, we performed a thorough analysis of CD28-mediated signaling events and functional responses with purified naïve T cells from Itk-/- mice and a highly controlled stimulation system. Data from this set of studies definitively demonstrate that CD28 costimulation functions efficiently in naïve CD4+ T cells in the absence of Itk. Second, in order to further study the functions of Tec kinases in vivo, we generated transgenic mouse lines expressing a kinase-dead (KD) mutant of Tec on the Itk-/-Rlk-/- background, hoping to study mice that are functionally deficient for all three Tec kinases. The results hint the importance of the Tec kinases in T cell development and/or survival. Finally, in order to identify potential transcriptional targets of Itk, we used microarray technology to compare global gene expression profiles of naïve and stimulated Itk-/- versus Itk+/- CD4+ T cells. This analysis provided a short list of differentially expressed genes in Itk-/- versus Itk+/- CD4 T cells, providing a starting point for further studies of Itk in T cell activation. Collectively, these studies clarified the role of Itk in CD28 signaling, revealed some unexpected aspects of Tec family kinases in T cells, and indicated potential targets of Itk-dependent signaling pathways in T cells.

A person is exposed to many pathogens throughout their lifetime, and with the resolution of each infection, there remains a pool of pathogen-specific immune cells that protect that person from re-infection with the same pathogen. However, there is a great deal of evidence to suggest that the pool of pathogen-specific memory cells can also participate in the immune response to future infections with unrelated pathogens. Many believe T cells to be cross-reactive in nature because of their interaction with self antigens during development in the thymus and their interaction with foreign antigens once in the periphery. There are many features of the interaction between a T cell and its ligand that facilitate this cross-reactive nature. Based on solved crystal structures, relatively few contacts are required for a stable interaction, and that interaction is often mediated by the flexible CDR3 loops of the T cell receptor that accommodate ligands of various structure. There is also evidence in the murine and human systems that subsets of virus-specific memory CD8 T cells take on an activated phenotype upon infection with an unrelated virus. In murine models, these memory T cell subsets could kill target cells, secrete several cytokines, and proliferate in response to a cross-reactive stimulation, suggesting that a cross-reactive T cell response could impact the outcome of a viral infection. In fact, upon heterologous infection, mice immune to a previous virus were often protected, having lower titers of the second unrelated virus, their epitope-specific and T cell receptor repertoires were often skewed, and they were more prone to immune-mediated pathologies. All of these observations coincided with the presence of cross-reactive T cell responses. Thus, we define heterologous immunity as changes in viral replication and the disease pathology associated with that viral infection as a result of the host's history of infection, and this can be mediated, in part, by cross-reactive CD8 T cell responses. Since many human viral infections are associated with a wide range of disease states, we questioned whether cross-reactive CD8 T cell responses occurred as commonly as they appeared to occur in the murine models and whether they influenced the outcome of such infections. Epstein-Barr virus (EBV) infects over 90% of the U. S. population and has a large genome with the capacity to encode a multitude of T cell epitopes. The first part of this thesis research focuses on the identification of cross-reactive CD8 T cell responses with specificity for known epitopes derived from EBV, a common human virus. We directed our study to HLA-A2-restricted responses because of the common expression of this MHC Class I allele in the U. S. population. This study resulted in the detection of cross-reactive responses with five different specificities that involved either the immunodominant lytic EBV-BMLF1280 epitope or the latent EBNA 3A596epitope. Three of the cross-reactive responses had specificity for epitopes derived from another unrelated, but common, human virus, influenza A virus (IV). Each of these cross- reactive responses had the potential to participate in the collective immune response to acute EBV infection. EBV is also well-suited as a model system to study heterologous immunity in humans, as infection at an early age is frequently asymptomatic, while the same infection during adolescence often results in an immune-mediated syndrome, infectious mononucleosis (IM). Since older individuals have presumably been exposed to more pathogens in their lifetime and, therefore, would have memory CD8 T cell pools with more extensive specificities, we hypothesized that acute EBV infection activated cross-reactive memory CD8 T cell responses that promoted the development of IM. In order to determine if the cross-reactive responses we identified above contributed to the immune response to acute EBV infection, we first screened the blood of IM patients for cross-reactive T cells with specificity for EBV-BMLFl280 and IV-M158. The total number of M1-specific T cells of 5 of 8 patients was increased at presentation with IM, which was suggestive of their specific activation during the EBV infection since a bystander mechanism would have resulted in 8 out of 8 patients having increased numbers of M1-specific T cells. Our hypothesis was further supported by the fact that we clearly detected cross-reactive T cells capable of recognizing both BMLF1 and M1 epitopes in the blood of 2 of the 5 IM patients with an augmented M1-specific T cell frequency. Furthermore, the M1-specific TCR repertoires of those two patients were dramatically skewed, which was an indication of cross-reactive M1-specific T cell expansions and, therefore, participation in the lymphoproliferation characteristic of IM. In addition, T cell lines derived from 3 out of 8 healthy donors with previous exposure to both viruses contained a subset of T cells that responded to both BMLF1 and M1 epitopes, suggesting that these cross-reactive cells are often maintained in memory. These cross-reactive T cells were cytotoxic and produced MIP-1β, IFNγ, and TNFα, functions which could potentially promote the symptoms of IM and, indeed, may have been contributed to the severe case of IM noted in one patient. The final part of this thesis research focused on defining the structure of the cross-reactive TCR that recognized both BMLF1 and M1 epitopes, which have only 33% sequence similarity. In addition, we examined the cross-reactive TCR repertoire organization of multiple individuals to determine the breath and, therefore, the likelihood that this cross-reactive T cell response will occur. These studies revealed that a wide range of Vα and Vβ families can mediate interaction with both epitopes and that the cross-reactive TCR repertoire was unique to each individual, relying heavily on the T cell clones present in that individual's private BMLF1- and M1-specific repertoires. We also observed an increased frequency of TCRs with longer CDR3 regions within the cross-reactive repertoire, which were often extended by non-bulky amino acid residues that could provide these TCRs with more flexibility in order. to accommodate the two different epitope structures. Given that we detected a cross-reactive T cell response with specificity for two immunodominant epitopes derived from two of the most common human viruses among people that share one of the most common MHC Class I alleles in the U. S. population, we predict that cross-reactive T cells are common components of human immune responses. The variability in the magnitude and specificity of each cross-reactive T cell response is dependent on each individual's unique history of infection and th,eir unique TCR repertoire, and such responses likely represent one of many factors that could explain the individual variability in disease severity associated with EBV and many other human viral infections.

Introdução: O crescente aumento do número de casos de AIDS em mulheres evidencia um progressivo aumento da Transmissão Vertical (TV) do HIV, gerando assim a necessidade de focalizarmos esforços no intuito de impedir ou minimizar este tipo de transmissão. Objetivos: avaliar as condutas profiláticas adotadas para redução da TV do HIV em serviços de referência pertencentes à 7ª CRS. Metodologia: Os dados foram obtidos por meio de pesquisa de campo retrospectiva com análise quanti-qualitativa dos formulários de cadastro, fichas de notificação e investigação compulsória, além de prontuários das gestantes e puérperas soro positivas para o HIV, no período de 01 de janeiro de 2010 a 01 de janeiro de 2013 mediante aprovação ética. Resultados: A população estudada compreendeu um universo de 22 mulheres notificadas como Gestante HIV +, dentre as quais 72% foram notificadas na cidade de Bagé, e destas, 9% foram notificadas em 2010, 27,3% em 2011 e 63,7% em 2012. A faixa etária de maior prevalência é de 15 a 24 anos (59%), 50% das mulheres não teve a via de parto nem a evolução da gravidez informada e 60% não tem registro de profilaxia no período intra-parto. Em 50% das fichas, não foi informado o uso da medicação pela criança nas primeiras 24h e 59% das notificações foram feitos por profissional enfermeiro. Pôde-se caracterizar com os dados coletados os aspectos individual, escolar e sócio-demográfico da 7ª CRS. Indo ao encontro das informações publicadas por Brasil (2013), este estudo evidencia uma sub-enumeração de casos no SINAN, pois apenas 63% das mulheres foram notificadas/cadastradas nos sistemas de informação. Considerações: As práticas de saúde adotadas pelos serviços estão de acordo Protocolo Vigente, no que se trata da administração da TARV a partir da 14ª semana de gestação, da escolha, sempre que possível da via de parto, a administração de medicamentos intra-parto, a inibição da lactação, o uso do ARV no bebê e o fornecimento de fórmula láctea. As medidas educativas necessárias para qualificar o atendimento realizado na região de saúde estudada relacionam-se principalmente à qualidade das informações das ações de saúde (alimentação dos bancos de dados dos sistemas), além de estimular a população em geral para o conhecimento do prénatal e das testagens para HIV. Para que pudéssemos trabalhar acerca deste tema, discutimos os resultados do estudo em equipe no SAIS de Bagé, e pensamos em algumas estratégias que nos aproxime dos demais profissionais da rede. Criamos um perfil em uma rede social, onde estão sendo postadas diariamente informações acerca dos temas: HIV, AIDS, DSTs, Hepatites Virais, medidas de prevenção, meios de transmissão, atividades do serviço, dados importantes, dentre tantas outras informações, sendo imprescindível relatar que todos os componentes da equipe tem acesso a senha, para que possa sentir-se ator no processo de educação da comunidade. Logo, serão propostas oficinas e fóruns, com os temas em questão para discutir com a comunidade local, com o apoio da equipe da 7ª CRS/RS. / The increasing number of AIDS cases among women reveals a progressive increase of Vertical Transmission (VT) of HIV, thus generating the need of focusing efforts in order to prevent or minimize this type of transmission. The aim of the study was to evaluate the prophylactic measures adopted to reduce HIV in TV reference belonging to the 7th CRS services. Data were obtained from retrospective research field with quantitative and qualitative analysis of the registration forms, reporting forms and compulsory research, and medical records of pregnant and postpartum serum positive for HIV in the period from January 1, 2010 to January 1, 2013 by ethical approval. The study population comprised a universe of 22 women reported / registered as HIV + Pregnant Women, among which 72% were reported in the city of Bage, while 09% reported in 2010, 27.3% in 2011 and 63.7% in the year 2012 the most prevalent age group is 15-24 years (59%), 50% of women had no way of delivery, or pregnancy outcome informed and 60% have no record of prophylaxis in intra period partum. In 50% of the chips, was not informed use of medication by the child in the first 24h and 59% of registrations / notifications were made by professional nurses. Could be characterized with the data the individual, school and socio-demographic aspects of CRS 7th. Going against the information published by Brazil (2013), this study highlights an under-enumeration of cases in SINAN because only 75% of women were reported / registered in information systems. Practices adopted by health services meet the Effective Protocol, it is the administration of ART from the 14th week of gestation, the choice where possible the mode of delivery, medication administration intra-partum, the inhibition of lactation, the use of ARVs in providing baby and milk formula. Educational measures to improve service held at the health region studied relate primarily to information quality of health (nutrition database systems), in addition to encouraging the general population to the knowledge of prenatal and testings for HIV. So we could work on this subject, discuss the results of the study team at SAIS Bage, and think of some strategies that bring us closer to the other networking professionals. We created a profile on a social network, where they are being posted daily information on the topics: HIV, AIDS, STDs, Viral Hepatitis, prevention, transmission media, service activities, important data, among many other information, is indispensable report that all team members have access to the password, so you can feel an actor in the process of educating the community. Soon, workshops and forums will be proposed, with the issues in question to discuss with the local community, with the support of the 7th CRS / RS staff.

Neisseria meningitidis (Nm) is one of the major causes of bacterial meningitis in the African meningitis belt (AMB). This organism is part of the genus Neisseria, which includes ten human restricted species, mostly harmless commensals of the nasopharynx; however, Nm is capable of causing invasive meningococcal disease. The transition from carriage to pathogenic state remains perplexing, and strict virulece factors have yet to be identified. It has been hypothesised that non-pathogenic Neisseria (NPN) carried asymptomatically in the oroopharynx could play a role in modulating carriage of Nm, and therefore, its likelihood of invasion. In chapter 3, the diversity of the genus was characterised within a collection of 46 034 nasopharyngeal samples obtained across the AMB: five different species were identified, with Nm and NPNs displaying inversely related risk factors fo carriage. Chapter 5 presents the whole genome sequence (WGS) analysis of 107 Neisseria isolates unclassified by other methods. This higher genetic resolution, complemented with the use of a novel speciation approach, revealed seven novel Neisseria species, mostly collected in African countries. The invasive potential may also be due to the presence of particular genetic factors in the meningococcal genome. Chapter 4 presents the WGS comparison of 23 carried and invasive serogroup A Nm collected in Chad during the 2011 meningitis epidemic. Isolates from both phenotypic groups were found to be part of the same bacterial populations; however, discrete clusters were identified, associated with distinct age groups. These results indicate that genomic analyses are essential to appropriately study Neisseria diversity, and that lower resolution methods have greatly underestimated the diversity of the genus in Africa. The identification of Nm clusters associated with certain niches and of the differences in carriage risk factors suggests that variation in the environment, including the presence of NPNs, may be key in modulating carriage of Nm.

The widespread use of antibiotics has led to dramatic increases in the incidence and severity of Clostridium difficile infection (CDI). No group of patients suffers more from CDI than the elderly. Nursing homes (NH) represent the perfect storm of a vulnerable group of frail elders living in confined communities. Nursing home residents suffer from increased morbidity and mortality from CDI and corresponding high rates of C. difficile colonization. Upwards of 40 to 50% of CDI current cases originate from NHs and the prevalence of colonization rates remain high within these facilities, with as many as half of the residents being colonized with C. difficile at any given time. One factor that has become of increasing interest and a target of preventive strategies is the human intestinal microbiome. A healthy, diverse microbiome interacts with the host immune system and contributes to pathogen resistance. In this investigation, we first examine elder specific variables to determine if the associated risks of CDI differ by home living environment (nursing home versus community-dwelling). We then go on explore the relationships of NH environment, frailty, nutritional status, and residents’ age with microbiome composition and potential metabolic function. Finally, we describe the C. difficile colonization patterns among elderly NH residents and the associated risk of colonization based on clinical variables and microbiome determinants. A better understanding of the microbiome’s contribution to C. difficile colonization will provide the basis for informing rational interventions and public health policies to better combat CDI in the nursing home.

Introdução: O crescente aumento do número de casos de AIDS em mulheres evidencia um progressivo aumento da Transmissão Vertical (TV) do HIV, gerando assim a necessidade de focalizarmos esforços no intuito de impedir ou minimizar este tipo de transmissão. Objetivos: avaliar as condutas profiláticas adotadas para redução da TV do HIV em serviços de referência pertencentes à 7ª CRS. Metodologia: Os dados foram obtidos por meio de pesquisa de campo retrospectiva com análise quanti-qualitativa dos formulários de cadastro, fichas de notificação e investigação compulsória, além de prontuários das gestantes e puérperas soro positivas para o HIV, no período de 01 de janeiro de 2010 a 01 de janeiro de 2013 mediante aprovação ética. Resultados: A população estudada compreendeu um universo de 22 mulheres notificadas como Gestante HIV +, dentre as quais 72% foram notificadas na cidade de Bagé, e destas, 9% foram notificadas em 2010, 27,3% em 2011 e 63,7% em 2012. A faixa etária de maior prevalência é de 15 a 24 anos (59%), 50% das mulheres não teve a via de parto nem a evolução da gravidez informada e 60% não tem registro de profilaxia no período intra-parto. Em 50% das fichas, não foi informado o uso da medicação pela criança nas primeiras 24h e 59% das notificações foram feitos por profissional enfermeiro. Pôde-se caracterizar com os dados coletados os aspectos individual, escolar e sócio-demográfico da 7ª CRS. Indo ao encontro das informações publicadas por Brasil (2013), este estudo evidencia uma sub-enumeração de casos no SINAN, pois apenas 63% das mulheres foram notificadas/cadastradas nos sistemas de informação. Considerações: As práticas de saúde adotadas pelos serviços estão de acordo Protocolo Vigente, no que se trata da administração da TARV a partir da 14ª semana de gestação, da escolha, sempre que possível da via de parto, a administração de medicamentos intra-parto, a inibição da lactação, o uso do ARV no bebê e o fornecimento de fórmula láctea. As medidas educativas necessárias para qualificar o atendimento realizado na região de saúde estudada relacionam-se principalmente à qualidade das informações das ações de saúde (alimentação dos bancos de dados dos sistemas), além de estimular a população em geral para o conhecimento do prénatal e das testagens para HIV. Para que pudéssemos trabalhar acerca deste tema, discutimos os resultados do estudo em equipe no SAIS de Bagé, e pensamos em algumas estratégias que nos aproxime dos demais profissionais da rede. Criamos um perfil em uma rede social, onde estão sendo postadas diariamente informações acerca dos temas: HIV, AIDS, DSTs, Hepatites Virais, medidas de prevenção, meios de transmissão, atividades do serviço, dados importantes, dentre tantas outras informações, sendo imprescindível relatar que todos os componentes da equipe tem acesso a senha, para que possa sentir-se ator no processo de educação da comunidade. Logo, serão propostas oficinas e fóruns, com os temas em questão para discutir com a comunidade local, com o apoio da equipe da 7ª CRS/RS. / The increasing number of AIDS cases among women reveals a progressive increase of Vertical Transmission (VT) of HIV, thus generating the need of focusing efforts in order to prevent or minimize this type of transmission. The aim of the study was to evaluate the prophylactic measures adopted to reduce HIV in TV reference belonging to the 7th CRS services. Data were obtained from retrospective research field with quantitative and qualitative analysis of the registration forms, reporting forms and compulsory research, and medical records of pregnant and postpartum serum positive for HIV in the period from January 1, 2010 to January 1, 2013 by ethical approval. The study population comprised a universe of 22 women reported / registered as HIV + Pregnant Women, among which 72% were reported in the city of Bage, while 09% reported in 2010, 27.3% in 2011 and 63.7% in the year 2012 the most prevalent age group is 15-24 years (59%), 50% of women had no way of delivery, or pregnancy outcome informed and 60% have no record of prophylaxis in intra period partum. In 50% of the chips, was not informed use of medication by the child in the first 24h and 59% of registrations / notifications were made by professional nurses. Could be characterized with the data the individual, school and socio-demographic aspects of CRS 7th. Going against the information published by Brazil (2013), this study highlights an under-enumeration of cases in SINAN because only 75% of women were reported / registered in information systems. Practices adopted by health services meet the Effective Protocol, it is the administration of ART from the 14th week of gestation, the choice where possible the mode of delivery, medication administration intra-partum, the inhibition of lactation, the use of ARVs in providing baby and milk formula. Educational measures to improve service held at the health region studied relate primarily to information quality of health (nutrition database systems), in addition to encouraging the general population to the knowledge of prenatal and testings for HIV. So we could work on this subject, discuss the results of the study team at SAIS Bage, and think of some strategies that bring us closer to the other networking professionals. We created a profile on a social network, where they are being posted daily information on the topics: HIV, AIDS, STDs, Viral Hepatitis, prevention, transmission media, service activities, important data, among many other information, is indispensable report that all team members have access to the password, so you can feel an actor in the process of educating the community. Soon, workshops and forums will be proposed, with the issues in question to discuss with the local community, with the support of the 7th CRS / RS staff.

abstract: Peptide microarrays are to proteomics as sequencing is to genomics. As microarrays become more content-rich, higher resolution proteomic studies will parallel deep sequencing of nucleic acids. Antigen-antibody interactions can be studied at a much higher resolution using microarrays than was possible only a decade ago. My dissertation focuses on testing the feasibility of using either the Immunosignature platform, based on non-natural peptide sequences, or a pathogen peptide microarray, which uses bioinformatically-selected peptides from pathogens for creating sensitive diagnostics. Both diagnostic applications use relatively little serum from infected individuals, but each approaches diagnosis of disease differently. The first project compares pathogen epitope peptide (life-space) and non-natural (random-space) peptide microarrays while using them for the early detection of Coccidioidomycosis (Valley Fever). The second project uses NIAID category A, B and C priority pathogen epitope peptides in a multiplexed microarray platform to assess the feasibility of using epitope peptides to simultaneously diagnose multiple exposures using a single assay. Cross-reactivity is a consistent feature of several antigen-antibody based immunodiagnostics. This work utilizes microarray optimization and bioinformatic approaches to distill the underlying disease specific antibody signature pattern. Circumventing inherent cross-reactivity observed in antibody binding to peptides was crucial to achieve the goal of this work to accurately distinguishing multiple exposures simultaneously. / Dissertation/Thesis / Doctoral Dissertation Biological Design 2014

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No. of bitstreams: 2 Tese - Lázaro Moreira Marques Neto - 2018.pdf: 4983908 bytes, checksum: 78504fbead82e1981e7577763889e31e (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-10-09 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The search for new adjuvants is the main goal in vaccinology. Along with this, understanding the impact of using nanoparticles as a delivery system and immunomodulator in vaccine systems directly impacts the development of new vaccines. In this work, we seek to study and elucidate the adjuvanticity of magnetic nanoparticles, as well as its immunogenicity and protection of the vaccine systems. Initially, a literature review was made seeking scientific bases that demonstrated the possibility of using metallic nanoparticles (MeNPs) as innate immune system stimulators. It was also sought to find elements in which metallic nanoparticles could aid in the generation Th1, Th17 and T CD8 type cellular response. From this review, it was verified that the magnetic nanoparticles, or with metallic ions, were able to stimulate the activation of costimulatory molecules (CD80, CD40 and CD86), to induce secretion of cytokines (IL-1, IL-6, IFN-γ and TNF-α) as well as the humoral immune response, but no work demonstrated whether these nanoparticles were able to induce cellular response. Consequently, in the second part of the study, tuberculosis was used as model to verify if a vaccine formulation with a magnetic nanoparticle of manganese ferrite combined with recombinant fusion protein would have the ability to induce a protective cellular immune response, without adding other adjuvants. The nanoparticle was coated with recombinant CMX fusion protein and BALB/c mice were vaccinated with this formulation, in protocol with three vaccinations with 21-day intervals. Subsequently, the vaccinated animals were infected with Mycobacterium tuberculosis (H37Rv) to evaluate the protection conferred by the vaccine. The results showed that the nanoparticle was able to generate cellular immune responses of Th1, Th17 and T CD8 types, depending on the route of inoculation (subcutaneous, intranasal and mixed). The most preeminent response was Tc1 which was recalled after infection was able to protect against the challenge with Mtb. In addition, there was no appearance of side effects or damage to organs of infected animals, demonstrating that the formulation is safe. Finally, the vaccine formulations with MeNPs, more specifically with manganese ferrite, demonstrate potential application in vaccinology, and may be applied in vaccine formulations to generate cellular immune response, but the route must be considered and in case of use other adjuvants it should consider the possible interaction of NP with the molecule and their ligand. / A busca por novos adjuvantes é um dos objetivos principais dentro da vacinologia. Juntamente com isso, entender o impacto do uso de nanopartículas como sistema de entrega e imunomodulador em sistemas vacinais impacta diretamente no desenvolvimento de novas vacinas. Nesse trabalho, buscamos estudar e elucidar a adjuvanticidade de nanopartículas magnéticas, bem como a imunogenicidade e proteção de sistemas vacinais utilizando essas nanopartículas. Inicialmente foi feito uma revisão da literatura buscando bases científicas que demonstrassem a possiblidade do uso de nanopartículas metálicas (MeNPs) como estimuladores do sistema imune inato. Buscou-se também encontrar elementos em que as nanopartículas metálicas pudessem auxiliar na geração de uma resposta celular do tipo Th1, Th17 e T CD8. A partir dessa revisão, verificou-se que as nanopartículas magnéticas, ou com íons metálicos, eram capazes de estimular a ativação de moléculas coestimuladoras (CD80, CD40 e CD86), induzir secreção de citocinas (IL-1, IL-6, IFN-γ e TNF-α) bem como a resposta imune humoral, mas nenhum trabalho demonstrou se essas nanopartículas eram capazes de induzir resposta celular. Consequentemente, na segunda parte do trabalho utilizou-se a tuberculose como modelo de estudo para verificar se uma formulação vacinal com uma nanopartícula magnética de ferrita de manganês combinada com proteína de fusão recombinante, teria capacidade indutora de resposta imune celular protetora, sem adição de outros adjuvantes. A nanopartícula foi recoberta com a proteína de fusão recombinante CMX e os camundongos BALB/c foram vacinados com essa formulação, em protocolo com três vacinações com intervalos de 21 dias. Posteriormente, os animais vacinados foram infectados com Mycobacterium tuberculosis (H37Rv) para se avaliar a proteção conferida pela vacina. Os resultados mostraram que a nanopartícula teve capacidade de gerar resposta imune celular dos tipos Th1, Th17 e T CD8, dependendo da via de inoculação (subcutânea, intranasal ou mista). Essa resposta foi principalmente do tipo Tc1 e foi capaz de proteger contra o desafio com Mtb. Adicionalmente, não houve qualquer aparecimento de efeito colateral ou danos em órgãos dos animais infectados, demonstrando que a formulação é segura. Por fim, as formulações vacinais com MeNPs, mais especificamente com ferrita de manganês, então demonstram potencial aplicação em vacinologia, podendo ser aplicada em formulações vacinais para gerar resposta imune celular, mas deve-se levar em conta a rota e, caso for utilizar outros adjuvantes complementares, deve-se pensar na possível interação da NP com o adjuvantes e seus ligantes.