Thèse de doctorat : Biologie moléculaire : Nancy 1 : 2007. / Titre provenant de l'écran-titre.

Kenya has experienced severe arboviral outbreaks of public health concern in the recent past, including yellow fever (YF), Crimean Congo hemorrhagic fever (CCHF), chikungunya, and Rift Valley fever (RVF) among others. Most of these infections are under diagnosed and hence neglected due to non-specific nature of their symptoms. Often they are mistaken for endemic tropical diseases such as malaria and typhoid infections and are only recognized during major outbreaks which result in adverse public health and economic consequences to the affected communities. Ongoing inter-epidemic surveillance in RVF virus hotspots in Kenya has indicated continued intense transmission of Bunyamwera virus (BUNV) in the absence or under low level activity of RVF virus. BUNV belongs to the genus Orthobunyavirus of the family Bunyaviridae. These are segmented RNA viruses whose members have the potential for genetic reassotment and/or drift. Recently, Ngari virus (NRIV), a natural reassortant virus associated with hemorrhagic fever was documented to have emerged from BUNV, which previously was not associated with such symptoms. However, the vectors that are involved in the maintenance and transmission of BUNV and NRIV are diverse and their role in virus maintenance/dynamics is poorly known. It is thus important to investigate the dynamics of BUNV and NRIV in selected transmission foci in an effort to understand their circulation better in order to be able to control and predict outbreaks. In this study, we determined the evolutionary and phenotypic diversity of BUNV and NRIV isolates previously obtained from vectors in parts of Kenya. We have provided genetic sequences of two BUNV and three NRIV isolates which contribute to addressing paucity of genetic sequences associated with this group of viruses. Phylogenetic analysis of these sequences in addition to other sequences in GenBank revealed evidence of geographic/temporal clustering that requires further investigation. Next, we demonstrated that plaque purified phenotypes of selected BUNV and NRIV isolates differ in in vivo growth kinetics and pathogenicity in mice, possibly related to specific mutations within the genome. The phenotypic changes and identification of mutations possibly associated with these changes support further investigation of specific mutations using site directed mutagenesis. In addition, we determined the competence of some of the mosquito species implicated in their transmission, Anopheles gambiae, Aedes aegypti and Culex quinquefaciatus and evaluated the dynamics of their transmission in these vectors. We conclude that Anopheles gambiae is likely a more competent vector for NRIV than Aedes aegypti and is a moderately competent vector for BUNV, which has implications for animal movement in malaria endemic areas where the vector is present. We also report evidence of BUNV transovarial transmission in both Aedes aegypti and Anopheles gambiae with the prevalence of transmission related to the ovarian cycle. Finally, we determined the level of human exposure to these viruses in the transmission foci. Orthobunyavirus-specific antibodies were detected by plaque reduction neutralization test in 89 (25.8%) of 345 persons tested. Multivariable analysis revealed age and residence in North Eastern Kenya as risk factors. In conclusion, we propose that acute febrile disease surveillance needs to be implemented in North Eastern Kenya. This study helps identify the virus strains/populations and the vector species that play a critical role in sustenance and transmission of BUNV and NRIV in different ecosystems in the country. All these are important in understanding virus circulation, potential for emergence and risk to populations in areas of circulation, and will help in making decisions for intervention and management. Generated sequence data in this study contributes to global phylogenetic characterization of Orthobunyaviruses worldwide and their molecular epidemiology. The study also shed light/improve our knowledge on the genetic stability or diversity and evolutionary trends of Orthobunyavirus strains in Kenya. / Thesis (PhD)--University of Pretoria, 2015. / Medical Virology / PhD / Unrestricted

Submitted by Valquíria Barbieri (kikibarbi@hotmail.com) on 2018-04-18T20:33:59Z No. of bitstreams: 1 DISS_2015_Belgath Fernandes Cardoso.pdf: 3865065 bytes, checksum: 3b7d79b4224429f58cb6ca5c3d96d7e9 (MD5) / Approved for entry into archive by Jordan (jordanbiblio@gmail.com) on 2018-04-27T17:29:40Z (GMT) No. of bitstreams: 1 DISS_2015_Belgath Fernandes Cardoso.pdf: 3865065 bytes, checksum: 3b7d79b4224429f58cb6ca5c3d96d7e9 (MD5) / Made available in DSpace on 2018-04-27T17:29:40Z (GMT). No. of bitstreams: 1 DISS_2015_Belgath Fernandes Cardoso.pdf: 3865065 bytes, checksum: 3b7d79b4224429f58cb6ca5c3d96d7e9 (MD5) Previous issue date: 2015-03-26 / CAPES / O gênero Orthobunyavirus, família Bunyaviridae, alberga arbovírus de importância médica. Estes estão envolvidos em epidemias de doença febril em áreas tropicais. No Brasil, o vírus Oropouche (OROV) é considerado o arbovírus mais frequente após o vírus da dengue (DENV). O objetivo deste estudo foi investigar a circulação de orthobunyavírus em Mato Grosso (MT). 529 amostras de soro obtidas entre outubro de 2011 e julho de 2012 de pacientes com doença febril aguda suspeita de dengue com até cinco dias do início dos sintomas em MT e 387 pools de mosquitos Cx quinquefasciatus capturados entre janeiro e abril de 2013 foram submetidos à nested RT-PCR para o segmento S de orthobunyavírus pertencentes ao sorogrupo Simbu. Amostras positivas foram testadas em pelo menos duas reações independentes e submetidas a sequenciamento nucleotídico para análise filogenética. Inoculou-se as amostras positivas em células vero. Dentre os 529 pacientes, 5 (0,94%) foram positivos para orthobunyavirus do sorogrupo Simbu por nested RT-PCR e isolamento viral. O vírus foi isolado de 3/8 pools. O segmento S do OROV foi identificado em cinco pacientes, quatro do sexo feminino, com 14-62 anos. Dois pacientes encontravam-se co-infectados com DENV-4. Estes pacientes são oriundos das cidades de Cuiabá (n=3), Várzea Grande (n=1) e Nova Mutum (n=1), todos residentes em área urbana, que apresentavam febre, cefaleia, dor retroorbital, mialgia, artralgia, prostação e náuseas. 8/387 pools de Cx. quinquefasciatus foram positivos para o segmento S do OROV por nested-RT-PCR, com taxa mínima de infecção (MIR) de 2,3 por 1000 mosquitos. As sequências obtidas do segmento S apresentaram 98% a 100% de homologia com o mesmo segmento das cepas do OROV verificadas no GenBank. A análise filogenética indica que as amostras de humanos e mosquitos pertencem ao subgenótipo Ia, similares a cepas do Pará obtidas de humanos, preguiças, Aedes (Ochlerotatus) serratus e Cx. quinquefasciatus. O genótipo I é o mais conservado e frequente no Brasil dentre os genótipos do OROV. Cx. quinquefasciatus, culicídeo de maior abundância em Cuiabá, é considerado vetor secundário do OROV em área urbana. Culicoides paraensis, principal vetor do OROV em áreas urbanas na região amazônica, não foi capturado neste estudo. Sorologia para o OROV foi identificada em residentes de cidades do Pará afetadas pela rodovia Cuiabá-Santarém e em primatas do Pantanal Sul-matogrossense, corroborando com a identificação do OROV em cidades do MT geograficamente interligadas por esta mesma rodovia. Infecções esporádicas por um orthobunyavirus do sorogrupo Simbu, possivelmente o OROV, foram identificadas em pacientes de MT, além de oito pools de Cx. quinquefasciatus em Cuiabá, indicando que este mosquito possui capacidade vetorial e pode estar envolvido no ciclo urbano de transmissão do vírus no estado. / The genus Orthobunyavirus, family Bunyaviridae, contains medically importante arboviruses. These are involved in epidemics of febrile illness in tropical areas. In Brasil, Oropouche virus (OROV) is considered the most frequent arbovirus after dengue virus (DENV). The aim of this study was to investigate the circulation of orthobunyaviruses in Mato Grosso (MT). 529 serum samples collected between October, 2011 and July, 2012 of patients with acute febrile illness suspected of dengue for up to five days of symptom onset from MT and 387 pools of Cx. quinquefasciatus mosquitoes captured between January and April, 2013, were subjected to nested RT-PCR for the segment S of orthobunyaviruses belonging to Simbu serogroup. Positive samples were tested in at least two independent reactions and subjected to nucleotide sequencing for phylogenetic analysis. Positive samples were inoculated in vero cells. Among the 529 patients, five (0.94%) were positive for serogroup Simbu by nested RT-PCR and viral isolation. The virus was isolated from 3/8 pools. The OROV segment S was identified in five patients, four females, with 14-62 years-old. Two patients were co-infected with DENV-4. These patients are from Cuiabá (n=3), Várzea Grande (n=1) and Nova Mutum (n=1), all residents in urban areas, presenting fever, headache, retroorbital pain, myalgia, arthralgia, prostration and nausea. 8/387 pools of Cx. quinquefasciatus were positive for OROV segment S by nested-RT-PCR with a minimum infection rate (MIR) of 2.3 per 1,000 mosquitoes. The segment S nucleotide sequences presented 98% to 100% of homology with the same segment of OROV strains available in GenBank. Phylogenetic analysis indicate the human and mosquito samples belong to genotype Ia, similar to strains obtained in Pará from humans, sloths, Aedes (Ochlerotatus) serratus and Cx. quinquefasciatus. The genotype I is the most conserved among OROV genotypes. Cx. quinquefasciatus, the most abundant culicidae in Cuiabá, is considered a secondary vector for OROV in urban areas. Culicoides paraensis, the main vector for OROV in urban areas in the Amazon region, was not captured in the study. Serology for OROV was identified in humans in cities of Pará affected by the Cuiabá-Santarém highway and in primates in South Pantanal, corroborating for the findings in cities of MT geographically linked by the same highway. Sporadic infections by an orthobunyavirus from Simbu serogroup, possibly OROV, were identified in patients from MT, also eight Cx. quinquefasciatus pools from Cuiabá, indicating that has vectorial capacity and may be involved in the urban cycle of virus transmission in the state.

Submitted by Cleide Dantas (cleidedantas@ufpa.br) on 2014-02-10T14:45:51Z No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Dissertacao_CaracterizacaoMolecularVirus.pdf: 2145926 bytes, checksum: a9fa4b1749574d6c532ca36612200d11 (MD5) / Approved for entry into archive by Ana Rosa Silva (arosa@ufpa.br) on 2014-04-22T13:52:31Z (GMT) No. of bitstreams: 2 Dissertacao_CaracterizacaoMolecularVirus.pdf: 2145926 bytes, checksum: a9fa4b1749574d6c532ca36612200d11 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Made available in DSpace on 2014-04-22T13:52:31Z (GMT). No. of bitstreams: 2 Dissertacao_CaracterizacaoMolecularVirus.pdf: 2145926 bytes, checksum: a9fa4b1749574d6c532ca36612200d11 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Previous issue date: 2007 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / As cepas do Virus Melao (VMEL), BE AR 8033 e BE AR 633512 foram isoladas de mosquitos Ochlerotatus (Ochlerotatus) scapularis, em Belém- PA (1955) e Alta Floresta do Oeste- RO (2000), respectivamente. Este trabalho teve como objetivo caracterizar molecularmente as cepas BE AR 633512 e BE AR 8033 e realizar estudos histopatológicos, bioquímicos e imunológicos comparativos em hamsters dourados (Mesocricetus auratus). Hamsters mostraram suscetibilidade às cepas do VMEL. A viremia em hamsters para BE AR 633512 ocorreu do 3º ao 6º dias pós-infecção (dpi.), e para a cepa BE AR 8033 ocorreu no 2º dpi. Anticorpos neutralizantes para ambas as cepas foram detectados a partir de 5 dpi., e se mantiveram até 30 dpi. As cepas testadas alteraram os marcadores bioquímicos AST, ALT e uréia, enquanto que a creatinina só apresentou alteração estatisticamente significante nos animais infectados com a cepa viral BE AR 633512, em comparação aos animais controles não infectados. Alterações histopatológicas foram observadas no SNC, fígado, rim e baço dos hamsters infectados pelas cepas do VMEL, sendo a infecção nesses órgãos confirmada por imunohistoquímica. A cepa BE AR 633512 foi mais virulenta e patogênica para hamsters que a cepa BE AR 8033. A análise genética dos genes N, Gn e Gc revelou que para os genes N e Gn, a cepa BE AR 8033 e do protótipo VMEL (TRVL 9375) são mais geneticamente relacionados. Para o gene Gc, a cepa BE AR 8033 é mais relacionada com a cepa BE AR 633512, sendo que esta última cepa apresentou maior variabilidade genética, principalmente no gene Gn com várias substituições de aminoácidos, mas as mutações no gene Gc provavelmente foram responsáveis pelo aumento da virulência e patogenicidade em hamsters. / The Melao Virus (MELV) strains, BE AR 8033 e BE AR 633512 were isolated from lots of Ochlerotatus (Ochlerotatus) scapularis mosquitoes, in Belém- PA (1955) and Alta Floresta do Oeste- RO (2000), Brazil, respectively. The aim of this study was to carry out molecular characterization of the MELV strains BE AR 633512 and BE AR 8033 and to describe the histopathologic, biochemistry and immunological changes in golden hamsters (Mesocricetus auratus). Hamsters showed susceptibility to these MELV strains. The viremia produced in hamsters by BE AR 633512 strain occurred between 3 and 6 days post-infection (dpi.), and for the BE AR 8033 occurred only in the 2nd dpi. Neutralizing antibodies for both strains were initially detected on the 5th dpi. and remained up to the 30 dpi. The biochemistry markers AST, ALT and blood urea nitrogen showed values statistically significants among inoculated animals with both VMEL strains, while creatinine value it was only altered by BE AR 633512 strain. Histopathologic changes were observed in the central nervous system, liver, kidney, and spleen of the hamsters, and infection was confirmed by immunohistochemical assay in all them. Strain BE AR 633512 caused more intense tissue damage showing increases virulence and pathogenicity than BE AR 8033 strain. The genetic analysis of the N, Gn and Gc genes revealed that for N and Gn genes, the BE AR 8033 and prototype strain (TRVL 9375) are genetically more closed related, and for the Gc gene, the BE AR 8033 and BE AR 633512 strains are more related. BE AR 633512 strain showed increased genetic variability, mainly in the Gn gene, were several aminoacid changes were observed, but the Gc mutations should be responsible for the increased virulence for hamsters.

Oropouche virus (OROV) is a medically important orthobunyavirus, which causes frequent outbreaks of a febrile illness in the northern parts of Brazil. However, despite being the cause of an estimated half a million human infections since its first isolation in Trinidad in 1955, details of the molecular biology of this tripartite, negative-sense RNA virus remain limited. The work presented in this thesis has re-determined the nucleotide sequences of OROV strain BeAn19991 (GenBank accession numbers: L, KP052850; M, KP052851 and S, KP052852), and demonstrates that the S segment is significantly longer than the published sequence with an additional 204 nucleotides at the 3' end. Data analysis revealed that there is a critical nucleotide mismatch at position 9 within the base-paired terminal panhandle structure of each genomic segment. Using a combination of deep sequencing and Sanger sequencing the complete genome sequences of 10 field isolates of OROV were also determined for the first time, and led to the identification of a novel OROV reassortant virus. Phylogenetic analysis of these sequences and of published sequences showed that there are two genotypes of OROV, rather than the four genotypes previously proposed. Further work led to the development of a T7-RNA polymerase-driven minigenome and virus-like particle (VLP) production systems for OROV; the information from these was subsequently used to develop a reverse genetics system for OROV. Using reverse genetics, OROV mutants that lack either the non-structural proteins NSm or NSs were generated. In vitro growth properties of the OROV mutant lacking NSm were indistinguishable from the wild-type virus, but the NSs mutant was attenuated in growth, particularly in interferon (IFN) competent cells. Further work demonstrated NSs as a viral IFN antagonist and that it's C-terminus is required for this activity. Interestingly, OROV is more resistant to IFN-α treatment than Bunyamwera virus, but this is not related to its NSs protein. The development of a reverse genetics system for OROV, which is the main human pathogen within the Simbu serogroup of orthobunyaviruses, will prove invaluable for future studies designed to further investigate the molecular pathogenesis of this virus and in the development of attenuated vaccine strains.

Le virus Schmallenberg (SBV) appartient au genre Orthobunyavirus, au sein de la famille des Bunyaviridae. Ce nouveau virus, découvert en 2011 au nord-ouest de l’Europe, affecte les ruminants domestiques. Il est responsable de signes cliniques discrets chez les adultes et de malformations congénitales chez les nouveau-nés. Ces travaux de thèse s’inscrivent dans les projets d’étude de la pathogenèse de l’infection à SBV et de l’épidémiologie de la maladie, dans le cadre d’un programme de recherche européen sur le virus. Ce manuscrit inclut de nouvelles données, telles les cinétiques de la virémie et de la séroconversion chez les ovins et caprins, après infection expérimentale par SBV. La possibilité d’infection par SBV par voie vaginale est démontrée expérimentalement chez la chèvre. Après infection expérimentale de chèvres gestantes entre 28 et 42 jours de gestation, une mortalité fœtale ou des lésions du système nerveux central des fœtus peuvent survenir. Enfin, la sensibilité de plusieurs espèces de ruminants sauvages et exotiques de parcs zoologiques vis-à-vis de SBV est démontrée pour la première fois. / Schmallenberg virus (SBV) belongs to the genus Orthobunyavirus in the family Bunyaviridae. This new virus was discovered in 2011 in Northwestern Europe in domestic ruminants. Infection by SBV is associated with mild clinical signs in adult and congenital malformations in the progeny. In the scope of the European research program on SBV in the pathogenesis and epidemiology areas, the works included in this thesis provide new data about SBV infection in livestock and wild and exotic ruminants. The kinetics of viremia and seroconversion after experimental SBV infection are described in sheep and goats. This manuscript includes evidence of SBV infection via vaginal route in goats. Experimental SBV infection in pregnant goats between 28 and 42 days of gestation can lead to death or central nervous system lesions in fetuses. Evidence of susceptibility to SBV in several species of wild and exotic ruminants kept in zoos is described for the first time.

Made available in DSpace on 2015-04-22T22:06:36Z (GMT). No. of bitstreams: 1 Dissertacao Final Gustavo Henrique.pdf: 11557113 bytes, checksum: 738eeceeb5f826798bc9ce2b09633f72 (MD5) Previous issue date: 2009-03-06 / FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas / Arbovirosis are currently recognized by the World Health Organization as a global problem. Most arbovirosis are summarized in diseases with acute and nonspecific symptoms such as fever, headache and muscle pain. Although self limited, these symptoms create relevant social and economic impacts. In Brazil, the arbovirosis caused by viruses belonging to the genus Alphavirus, Flavivirus and Orthobunyavirus and are the main cause of outbreaks or epidemics. This study aimed to study the circulation of arbovirus mayaro, venezuelan equine encephalitis virus, yellow fever virus, Saint Louis encephalitis virus and oropouche virus in a rural and an urban area of the State of Amazonas, Brazil. Therefore, the serology for detection of immunoglobulin G was used to assess the prevalence of antibodies against these viruses in 335 residents of a rural community in the state of Amazonas and PCR was used to assess the incidence of these viruses in 250 samples collected in urban area of Manaus. The results for serology suggest the movement of mayaro virus in the rural community. The seroprevalence detected in the samples was 41.5%. There was no significant relationship to risk for mayaro infection between genders (p value = 0.7760) or between age groups (p value = 0.9422). The positive serology detected among 39 children younger than 10 years indicates a recent infection. The factors of protection against mayaro infection detected were the use of mosquito net (p value = 0.0119) and the presence of animals in surrounding (p value = 0.0407). The risk factors identified for mayaro infection were the location of residence in towns near the forest (p value <0.0001) and presence of toilet in or near the home (p value = 0.0415). The serological results suggest that infection with mayaro occurred less than 10 years in the vicinity of residences analyzed. Molecular analysis of the samples collected in the urban area of Manaus not detected genomic fragments of arboviruses. Factors such as low viremia at the time of blood collection and storage of serum samples may have contributed to the non-detection of genomic fragments. However, the protocol for the detection of genomic fragments of arboviruses based on the PCR technique is already used in research centers and surveillance of Fundação de Medicina Tropical do Amazonas FMTAM and Instituto Leônidas e Maria Deane ILMD/FIOCRUZ. / As arboviroses são atualmente reconhecidas pela Organização Mundial da Saúde como um problema global. A maioria das arboviroses resume-se em afecções com sintomatologias agudas e inespecíficas, como febre, dores de cabeça e dores musculares. Embora sejam auto limitados, tais sintomas geram impactos sociais e econômicos relevantes. No Brasil, as arboviroses provocadas pelos por vírus pertencentes aos gêneros Alphavirus, Orthobunyavirus e Flavivirus são as principais causadoras de surtos ou epidemias. O presente estudo teve como objetivo estudar a circulação dos arbovírus mayaro, vírus da encefalite eqüina venezuelana, vírus da febre amarela, vírus da encefalite de Saint Louis e vírus oropouche em uma área rural e uma área urbana do Estado do Amazonas, Brasil. Assim sendo, a sorologia para detecção de imunoglobulinas G foi utilizada para avaliar a prevalência de anticorpos contra tais vírus em 335 moradores de uma comunidade rural do Estado do Amazonas e a PCR foi utilizada para avaliar a incidência de tais vírus em 250 amostras coletadas na área urbana de Manaus. Os resultados encontrados para a sorologia sugerem a circulação do vírus mayaro na comunidade rural. A soroprevalência detectada nas amostras foi de 41,5%. Não houve relação estatisticamente significativa de risco para a infecção por mayaro entre os gêneros (p valor=0,7760) ou entre as faixas etárias (p valor=0,9422). A sorologia positiva detectada entre 39 crianças menores de 10 anos indica uma infecção recente. Os fatores de proteção contra a infecção por mayaro detectados foram o uso de mosquiteiro (p valor=0,0119) e a presença de animais no peridomicílio (p valor=0,0407). Os fatores de risco detectados para a infecção por mayaro foram a localização do domicílio em vilas próximas à floresta (p valor<0,0001) e a presença de toalete dentro ou próximo ao domicílio (p valor=0,0415). Os resultados sorológicos sugerem que a infecção por mayaro ocorreu há menos de 10 anos nas proximidades das residências analisadas. A análise molecular das amostras coletadas na zona urbana de Manaus não detectou fragmentos genômicos de arbovírus. Fatores como baixa viremia no momento da coleta de sangue e estocagem das amostras de soro podem ter contribuído para a não detecção dos fragmentos genômicos. Contudo, o protocolo de detecção de fragmentos genômicos de arbovírus baseado na técnica de PCR está em uso nos centros de pesquisa e vigilância epidemiológica da Fundação de Medicina Tropical do Amazonas FMTAM e do Instituto Leônidas e Maria Deane ILMD/FIOCRUZ.

Submitted by Edisangela Bastos (edisangela@ufpa.br) on 2013-06-03T19:14:07Z No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Tese_CaracterizacaoRespostaImune.pdf: 1244558 bytes, checksum: c43d999fb552fb479140d57b56fda3a4 (MD5) / Approved for entry into archive by Ana Rosa Silva(arosa@ufpa.br) on 2013-06-04T14:13:21Z (GMT) No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Tese_CaracterizacaoRespostaImune.pdf: 1244558 bytes, checksum: c43d999fb552fb479140d57b56fda3a4 (MD5) / Made available in DSpace on 2013-06-04T14:13:21Z (GMT). No. of bitstreams: 2 license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Tese_CaracterizacaoRespostaImune.pdf: 1244558 bytes, checksum: c43d999fb552fb479140d57b56fda3a4 (MD5) Previous issue date: 2011 / IEC - Instituto Evandro Chagas / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / A tese aqui apresentada trata-se do primeiro estudo em nível mundial que pesquisa e caracteriza a resposta imune citocínica em infecções humanas pelo Orthobunyavirus Oropuche. Como metodologia para o alcance dos objetivos aqui apresentados foi utilizado um total de 320 amostras de soros humanos, onde 60 destas foram provenientes de Banco de Sangue (Controle negativo) e 260 foram obtidas mediante dois surtos do Vírus Oropouche nos Estados do Pará e Amapá (Brasil), sendo estas últimas divididas em oito subgrupos para obtenção dos dados com exatidão. Nas amostras coletadas foram realizadas análises dos dados clínicos/sintomatologia através dos prontuários, dados sorológicos através da titulação de anticorpos por Inibição da Hemaglutinação (IgM/IgG) e detecção do nível de citocinas plasmáticas por citometria de fluxo a qual permitiu a descrição técnica da dosagem de citocinas possibilitando ainda a análise de frequência de baixos e altos produtores de citocina. Os dados obtidos permitiram observar as variáveis e o comportamento das assinaturas de citocinas expressas pelos pacientes mediante a confirmação sorológica do vírus, bem como o comportamento destes analitos séricos quando da presença de sintomas específicos como febre, calafrios, cefaléia e tontura, permitindo assim que se chegasse à conclusão que a) existe um padrão na síntese de citocinas pró-inflamatórias e reguladoras; b) observa-se um balanço no perfil da resposta imune entre citocinas pró-inflamatórias (Th1) e moduladoras (Th17); c) a infecção pelo Vírus Oropouche altera a produção das citocinas nos indivíduos; d) os resultados mostram também que ao comparar os indivíduos Não respondedores com os Respondedores precoces, houve aumento da IL-1β e diminuição da IL-12; Não respondedores com Respondedores tardios, houve diminuição da IL-8, e aumento da IFN-α, IL-23 e IL-17; Não respondedores comparados com Respondedores precoces ocorreram o aumento de IL-4 e IFN-; Já quando comparado Respondedores precoces e respondedores tardios houve diminuição de IFN-α e IL-6; Respondedores precoces de forma geral apresentaram diminuição da IL-10 e Respondedores tardios apresentaram aumento da IL-5; e) Os resultados mostram ainda a expressão de IL-5 em pacientes que manifestaram os sintomas específicos para a infecção pelo Oropouche (febre, calafrios, cefaléia e tontura), sugerindo este sinal estar associado diretamente à patogênese do vírus; f) há a necessidade da complementação desta pesquisa com mais estudos como àqueles relacionados com a expressão de quimiocinas. / This thesis is the first global study that researches and analyzes the immune response of cytokine in human infections by Orthobunyavirus Oropuche virus. The study used 320 samples of human serum. Sixty were from the Blood Bank (negative control) and 260 were obtained from two outbreaks of the Oropouche virus in the State of Pará and Amapá (Brazil). The latter was divided into 8 subgroups for better data accuracy. The collected samples were analyzed for clinical data/symptoms with serologic testing by titration of antibodies by the hemagglutination inhibition (IgM/IgG) and the detection cytokines plasma levels by flow cytometry. This allowed for the technical description of cytokine. The data obtained allowed for the observation of the characteristics and the behavior of the cytokines signatures expressed by patients by the presence or not of the virus. This also allowed for the observation of changes to serum through the presence of specific symptoms such as fever, chills, headache and dizziness. This led to the following conclusions a) there is a pattern in the synthesis of pro-inflammatory and regulatory cytokines; b) there is a balance in the profile of the immune response between pro-inflammatory cytokines (Th1) and modulators (Th17); c) an infection by the Oropouche virus alters the production of cytokines in individuals; d) the results also show that whem comparing individuals no responders with early responders, there was an increase of IL-1β and decreased IL- 12; no responders with late responders, there was a decrease of IL-8, and increased IFN-α, IL-23 and IL-17; No responders occurred early responders compared with the increase IL-4 and IFN-g; However, when compared early responders and late responders, decreased IFN-α and IL-6; early responders generally showed a decrease in IL-10 and late responders showed an increase in IL-5; e) The results also show the expression of IL-5 in patients who showed symptoms specific for Oropouche infection (fever, chills, headache and dizziness), suggesting this signal to be directly associated with pathogenesis of the virus; f) there is a need to complement this research with more studies such as those related to the expression of chemokines.