Introdução: a coqueluche é uma doença causada pela Bordetella pertussis (BP), sendo mais frequente e grave em lactentes menores de um ano de idade. Com a introdução da vacina, houve redução na incidência mundial da doença, porém nos últimos 10 anos observa-se uma recrudescência. Pode apresentar-se de forma menos característica em lactentes, especialmente antes do final do esquema vacinal para o primeiro ano de vida. O quadro clínico, nesses pacientes, pode ser semelhante ao das infecções por vírus respiratórios (VR) que são os agentes etiológicos mais frequentes nas infecções de vias aéreas, nessa faixa etária. São necessários estudos que avaliem a importância da pesquisa de VR em lactentes com suspeita clínica de coqueluche. Objetivos: em lactentes com suspeita de coqueluche: identificar as prevalências de BP, VR e codetecções; analisar e comparar as características clínicas e a evolução, segundo a etiologia identificada e analisar o impacto do diagnóstico etiológico sobre o uso de macrolídeos. Métodos: estudo de coorte prospectivo, com crianças menores de um ano de idade, hospitalizadas com suspeita clínica de coqueluche entre junho de 2014 e junho de 2016 e submetidas à pesquisa etiológica para identificação de BP (\"swab\" de nasofaringe para cultura e/ou PCR) e pesquisa de VR (aspirado de nasofaringe para imunofluorescência indireta). Dados clínicos, demográficos e evolutivos foram coletados com o preenchimento de protocolo clínico-laboratorial padronizado. Resultados: no período de estudo foram analisados 59 lactentes. Em 18 (30,5%) houve identificação de BP, em 23 (39%) de algum vírus respiratório. Em quatro (7%), houve codetecção de BP e algum VR. O vírus mais frequentemente identificado foi o VSR (73%). As características com maior sensibilidade para o diagnóstico de infecção por BP foram tosse seguida de cianose e ser filho de mãe não vacinada com dTpa. Sibilos e desconforto respiratório apresentaram alta sensibilidade para a identificação de VR. Na análise bivariada apresentaram maior chance de infecção por BP: menor idade (OR = 1,86), ausência de febre (OR = 4,9), não ser vacinado para coqueluche (OR = 4,4), leucocitose superior a 20.000/mm3 (OR = 5,4), linfocitose superior a 10.000/mm3 (OR = 4,0) e de infecção por VR: sibilos (OR = 4,33). Após o ajuste para confundidores, os maiores preditores para BP de forma independente foram: ausência de sibilos (OR =5,7) e leucocitose superior a 20.000/mm3 (OR = 5,38). O número de pacientes com codetecção não permitiu a análise comparativa de gravidade com aqueles com agente único. Em apenas um paciente o resultado da pesquisa viral positiva resultou em suspensão de macrolídeo. Conclusão: além da BP, os VR também foram etiologias frequentes nos lactentes com suspeita clínica de coqueluche, além de casos de codetecção de BP e VR. Foram identificadas características clínicas/laboratoriais sugestivas, porém não patognomônicas das etiologias identificadas o que corrobora a necessidade da pesquisa etiológica para VR, nessa situação clínica / Introduction: Pertussis is a disease caused by Bordetella pertussis (BP), being more frequent and severe in infants less than one year old. After vaccine introduction, there was a reduction in the global incidence of the disease, but in the last ten years there was a resurgence. It may present less characteristically in infants, especially before the end of the vaccine scheme for the first year of life. The clinical picture in these patients may be similar to that of respiratory virus infections (VR), which are the most frequent etiologic agents in airway infections in this age group. Studies is necessary to evaluate the importance of RV research in infants with clinical suspicion of pertussis. Objectives: In infants with suspected pertussis: identify the prevalence of BP, VR and codetections; analyze and compare the clinical characteristics and evolution according to the identified etiology and analyze the impact of the etiological diagnosis on the use of macrolides. Methods: A prospective cohort study with children under one year of age hospitalized with suspected clinical pertussis between June 2014 and June 2016 and submitted to etiological research to identify BP (nasopharynx swab for culture and/or PCR) and VR (nasopharyngeal aspirate for indirect immunofluorescence). Clinical, demographic and evolution data were collected with the completion of a standardized clinical-laboratory protocol. Results: During the study period, 59 infants were analyzed. In 18 (30.5%) there was identification of BP, in 23 (39%) of some respiratory virus. In four (7%), there was BP detection and some RV. The virus most frequently identified was RSV (73%). The characteristics with greater sensitivity for the diagnosis of BP infection were cough followed by cyanosis and the mother\'s non-vaccinated dTpa. Wheezing and respiratory distress presented high sensitivity for RV identification. In the bivariate analysis they presented a greater chance of BP infection: lower age (OR = 1.86), absence of fever (OR = 4.9), not being vaccinated for pertussis (OR = 4.4), leukocytosis higher than 20,000/mm3 (OR = 5.4), lymphocytosis greater than 10,000/mm3 (OR = 4.0) and RV infection: wheezing (OR = 4.33). After adjustment for confounders, the largest predictors for BP independently were: no wheezing (OR = 5.7) and leukocytosis higher than 20,000/mm3 (OR = 5.38). The number of patients with codetection did not allow the comparative analysis of severity with those with single agent. In only one patient, the result of positive viral research resulted in macrolide suspension. Conclusion: In addition to BP, RVs were also frequent etiologies in infants with clinical suspicion of whooping cough, as well as cases of BP and VR codetection. Clinical/laboratory characteristics suggestive, but not pathognomonic, of the identified etiologies have been identified, which corroborates the need for etiological research for RV in this clinical situation

A Bordetella pertussis é a bactéria causadora da coqueluche, doença infectocontagiosa que tem reemergido em diversos países apesar das altas coberturas vacinais. Como os indivíduos mais afetados são crianças menores de seis meses de idade, a vacinação de gestantes com uma dose de vacina adsorvida contra tétano, difteria e coqueluche (B. pertussis acelular - dTpa) foi proposta. O objetivo é promover a transferência de altos títulos de anticorpos maternos ao feto e recémnascido, resultando na sua proteção até que o seu esquema vacinal esteja concluído. Esta estratégia é empregada em diversos países e no final de 2014 foi introduzida no Brasil. Entretanto, pouco se sabe sobre sua eficácia, efetividade e interação com a posterior vacinação da criança, que é realizada com a formulação de células inteiras (DTP), enquanto outros países substituíram-na pela formulação acelular (DTPa). Sabe-se que a vacinação materna induz anticorpos que ajudam a proteger recém-nascidos da infecção, mas podem, também, neutralizar a vacinação da criança, diminuindo a eficiência vacinal, além de promover a transferência de antígenos para o feto, induzindo desvio do perfil de resposta. Esses aspectos são particularmente importantes e devem ser investigados quando novos programas de vacinação materna são implantados. Sendo assim, o objetivo do presente estudo foi avaliar a resposta celular contra B. pertussis em parturientes e neonatos no dia do parto, e em lactentes após vacinação completa contra a coqueluche com DTP no primeiro ano de vida, comparando-se o grupo proveniente de mães que foram vacinadas durante a gestação com as que não receberam a dose de reforço. As células mononucleares do sangue periférico e de cordão umbilical dos participantes foram isoladas e estimuladas com antígeno bacteriano ou antígeno policlonal em cultura. Após o tempo estipulado, verificou-se o perfil de expressão gênica por qPCR nas células e a concentração de citocinas de interesse em sobrenadante, por meio de citometria de fluxo. Os dados obtidos mostram que a vacina na gestação induz resposta celular favorável à proteção contra infecção em gestantes, com produção de citocinas de perfil Th1 após estímulo antigênico, enquanto que nos neonatos poucas citocinas estavam acima dos limites de detecção. Quando se comparou a resposta de lactentes nascidos de mães vacinadas ou não durante a gestação, não foi encontrada diferença significativa nos níveis de citocinas produzidos, sugerindo que os anticorpos maternos presentes durante o desenvolvimento fetal e os primeiros meses de vida das crianças não interferiu no processo de reconhecimento e geração de resposta específica. Apesar do número amostral limitado, este trabalho mostra um panorama da interação da vacinação materna com a resposta celular à vacinação da criança no seu primeiro ano de vida, mostrando que a princípio a dose de reforço com dTpa durante a gestação é capaz de gerar resposta celular protetora nas gestantes, prevenindo-as de transmitir a doença a seus filhos, e a resposta humoral produzida não interfere na geração de resposta celular de seus filhos após o seu próprio esquema de imunização. / Bordetella pertussis is the bacterial agent of whooping cough, an infectious and contagious re-emerging disease despite high vaccine coverage. As the most affected are children younger than six months of age, a dose of tetanus/diphtheria/pertussis vaccine (acellular pertussis - Tdap) in pregnancy was proposed. It is aimed to promote the transfer of high maternal antibodies\' titres to the foetus and newborns, resulting in their protection until their own vaccination scheme is completed. This strategy is present in several countries, and it was implemented in Brazil by the end of 2014. However, little is known about its efficacy, effectiveness and interaction with the subsequent child\'s vaccination, which comprehends the whole-cell formulation (DTP), while other countries replaced it for the acellular version (DTaP). Maternal vaccination induces antibodies that would protect newborns from infection, but they can also neutralize the effect of the child vaccination, reducing the vaccine efficiency. Besides, it can promote the transfer of antigens to the foetus, which can induce deviation in the response pattern. These aspects are particularly important and should be investigated when new maternal vaccination programs are implemented. Therefore, the aim of this project was to evaluate the cellular response against B. pertussis in women at labour and their neonates, as in young children after complete pertussis vaccination by the first year of life, comparing groups from vaccinated and non-vaccinated women during pregnancy. Peripheral blood and cord blood mononuclear cells were isolated and stimulated with bacterial or polyclonal antigen in culture. After the stipulated time the gene expression profile and cytokine concentration were evaluated by qPCR and flow cytometry. Data from the present study show that pregnancy vaccination inducing favourable cellular response for protection against infection in pregnant women, with Th1 cytokines\' production upon antigenic stimulus, while in newborns few cytokines were detected above the detection limit. When comparing children\'s response, born either from vaccinated or unvaccinated mothers, cytokine levels were not significantly different, suggesting that maternal antibodies present during foetal development and first months of life do not interfere with recognition and cellular response generation to vaccination. Despite limited sample size, this work shows a broader view of interaction between maternal and child vaccination, showing that the Tdap boost dose during pregnancy may be able to generate protective response in women, preventing them to transmit the disease to children, and the induced humoral response may not interfere in generating cellular response in children after their own vaccination.

Introdução: A coqueluche é caracterizada por tosse paroxística, pode levar menores de um ano de idade ao óbito, deixar seqüelas e exacerbar quadros respiratórios crônicos. A imunidade após a doença ou vacina não é para toda a vida. Nos países desenvolvidos, apesar de altas coberturas vacinais e do controle da doença entre as décadas de 50 e 80, desde o final dos anos 80 é observado o aumento dos casos em adolescentes, adultos e lactentes, sendo indicado o reforço vacinal para adolescentes e adultos. No Brasil a doença aparenta estar sob controle, mas há um estudo teórico que demonstra a possibilidade de aumento dos casos. Objetivo: Avaliar novas estratégias de reforço vacinal contra a coqueluche no município de São Paulo. Metodologia: Desenvolvimento de um modelo matemático determinístico, dinâmico e dependente da idade dos indivíduos. Simulações com o esquema vacinal atual e: (i) novo reforço aos 12 anos com coberturas vacinais de 10%, 35%, 45% e 70%; (ii) reforços aos 12 anos e aos 20 anos de idade, com 35% e 70% de cobertura, respectivamente. Introdução de contato heterogêneo da população com o uso de uma matriz de contato. Fontes dos dados: Banco de dados do Sistema Único de Saúde (DATASUS), Centro de Vigilância Epidemiológica da Secretaria de Estado da Saúde de São Paulo e a literatura nacional e internacional. Uso dos programas Berkeley Madonna® para resolução das equações diferenciais e Microsoft Excel® para o cálculo da matriz de contato e das forças de infecção. Realização de teste de sensibilidade do modelo. Resultados: A vacinação com cobertura de 10% aos 12 anos de idade reduziu os casos entre os próprios adolescentes (10 a 19 anos); com cobertura de 35%, 45% e 70% reduziu os casos em 59%, 65% e 73%, respectivamente; a vacinação em conjunto aos 12 anos com cobertura de 35% e aos 20 anos com cobertura de 70% reduziu 62% dos casos. Conclusões: Há benefício ao vacinar os adolescentes, inclusive com baixa cobertura vacinal, portanto tal estratégia demonstra-se promissora para o controle da coqueluche. Não houve ganho ao acrescentar apenas um reforço para os adultos (20 anos). Os resultados são concordantes com o que há na literatura e permitiram um primeiro panorama para auxiliar na abordagem do problema. Estudos com diferentes estratégias de vacinação de adultos e estudos de custo-benefício são recomendados. / Background: Whooping cough is a respiratory tract infection caused by Bordetella pertussis and characterized by paroxysmal cough that usually causes complications for infants, including death, and for people with chronic respiratory diseases. Immunity against pertussis after infection or vaccination is not everlasting. Despite of high childhood immunization coverage and the disease control from the 50's to 80's, since late 80's developed countries notified high levels of pertussis in adolescents and adults. This reappearance has not being detected in Brazil yet, but at least one formal study has demonstrated the possibility of this change in the next years. Objective: Evaluating new pertussis vaccine's booster for adolescents and adults in São Paulo city. Methods: Development of a deterministic, compartmental and age-dependent model accounting for immunity waning. The data was retrieved from literature, Surveillance Center of the State of São Paulo (CVE), and the Brazilian national health data system (DATASUS). Data manipulation used Berkeley Madonna® and Microsoft Excel®. Vaccination strategies included the current vaccination scheme, plus (i) 10%, 35%, 45% or 70% vaccine coverage for those at the age of 12 and (ii) both 35% and 70% vaccine coverage at the ages 12 and 20, respectively. The Who Acquire Infection from Whom (WAIFW) matrices' method was used to assume age related transmission rates. Sensitivity analysis was performed. Results: Booster vaccination for 12 years youths, at 10% coverage, yields disease reduction only among adolescents (10 to 19 years); coverage up to 35% yields disease reduction for all ages; at 35%, 45% and 70% coverage, the reduction achieves 59%, 65% and 73%. Booster vaccination at 12 and 20 years, with coverage at 35% and 70% respectively, yields 62% cases reduction. Discussion: Results suggest that adolescent's vaccine booster could reduce pertussis occurrence for all ages, including infants, as also demonstrated by other authors. In contrast, only one vaccine booster for adults (20 years) achieves insignificant results. In conclusion, we have been able to demonstrate that, in São Paulo, the adolescent vaccine booster strategy is a promising police to further reduce whooping cough occurrence. However, cost effective analysis and other adults' vaccination strategies are recommended.

In 2002, a proposal was submitted to divert water from the confluence of the Guadalupe and San Antonio Rivers to San Antonio, Texas. To investigate the potential impacts of diverting water from the Guadalupe Estuary, my research focused on the foraging ecology of the crane and population ecology of the blue crab, a documented crane food. During winters 2004-2005 and 2005-2006, I examined diets and optimal foraging patterns of territorial adult cranes at Aransas National Wildlife Refuge, Texas. To identify foods of greatest benefit to cranes, I used currencies of optimization to evaluate foraging gain, cost, and efficiency. Foraging benefit differed among foods, depending on the foraging currency used and resource targeted (e.g., energy). Wolfberry fruit, snails, and insects were consumed in the highest quantities, required the least foraging effort, and were generally associated with the highest foraging efficiency. Blue crabs and clams were important sources of protein and biomass. During September 2003-October 2005, I used novel artificial settlement substrates and modified methods of standard deployment to investigate the spatio-temporal patterns of blue crab terminal settlement and recruitment rates. Monitoring rates in shallow bay habitat and ponds of the interior salt-marsh revealed megalopal crabs were developmentally advanced when arriving to study sites and the extent to which young crabs infiltrated the salt-marsh increased with age. Such findings suggested sites represented optimal terminal settlement habitat and consequently critical nursery habitat. Model selection indicated water temperature before and during embryonic development was the best predictor of megalopal settlement, whereas juvenile recruitment was most influenced by recent precipitation. I studied the size-specific abundance patterns of blue crabs in and around mature salt-marsh. Using drop-trapping and throw-trapping methods, I collected monthly samples in several habitats during October 2004-March 2006. Interior-marsh habitats contained fewer but larger crabs than bay habitat. Crabs contributing greatest to biomass were smallest (11-30 mm carapace-width) in bay habitat, larger (31-80 mm) along interior-marsh pond edges, and largest (41-130 mm) in interior-marsh open water. Model selection revealed crab density was most influenced by micro-site characteristics (habitat, water column structure type and structural complexity). Overall, shallow bay provided important nursery habitat for young blue crabs and interior marsh ponds were important for dispersing juvenile and adult crabs.

Whooping cough (pertussis) is a highly contagious respiratory infection caused by Bordetella pertussis. It becomes widespread among adolescent and adults as well as infants. Although availability of effective pertussis vaccines seems to decrease the incidence of the disease, B. pertussis circulation in population has not been eliminated. It is thought that the antigenic drifts in major protective antigens and continued circulation of B. pertussis strains will result in gradual loss of the efficacy of the current pertussis vaccines. Therefore, development of more effective acellular pertussis vaccines with conserved protective proteins is a convenient strategy to provide a better protection against whooping cough. In this study, immune responses against putative peptidyl-prolyl cis-trans isomerase (PPIase) which was shown to be immunogenic in B. pertussis for the first time by our immunoproteome group and FimX whose expression was found higher in our local Saadet strain were determined in mice. The genes encoding FimX and putative PPIase were amplified by PCR, cloned into pGEM&reg / -T Easy vector and sequenced. The genes were then introduced into pET-28a (+) vector and they were expressed in Escherichia coli BL21(DE3) cells. The recombinant proteins were purified by His-tag affinity chromatography and dialyzed. After Western blot analyses, 20 &micro / g and 80 &micro / g recombinant FimX and 80 &micro / g recombinant putative PPIase were used to immunize BALB/c mice (16-18 g) at day 0 and 21. The mice were challenged intranasally with 2.5 x 109 live B. pertussis Saadet cells. Before second immunization and challenge, the sera were collected to carry out ELISA for measurement of serum-specific IgG levels. According to ELISA results, IgG levels in the mice immunized with 20 &micro / g and 80 &micro / g recombinant FimX were found significantly higher than in control groups at both first and second vaccinations (p&lt / 0.01). On the other hand, immunization with 160 &micro / g recombinant putative PPIase provided a significant increase in IgG level (p&lt / 0.05) only at second vaccination. The lungs of the mice were removed at day 2, 5, 8 after challenge and bacterial colonization was determined. No significant decrease in bacterial colonization was observed in the lungs of the mice immunized with 20 &micro / g and 80 &micro / g recombinant FimX and 80 &micro / g recombinant putative PPIase with respect to control groups. After respiratory challenge and second immunization (at day 30) with 20 &micro / g and 80 &micro / g recombinant FimX, the spleens of the mice were removed and a spleen cell culture was obtained. Supernatants were collected after induction of the cells with the recombinant protein and cytokine ELISA was carried out to measure IFN-&gamma / level. No significant difference was observed between control and vaccinated mice in terms of IFN-&gamma / production.

Bordetella pertussis e B. parapertussis são agentes etiológicos da coqueluche. A IL-12 liga a imunidade inata e adaptativa. Investigamos alguns mecanismos que controlam a síntese de IL-12 em macrófagos medulares murinos (MfDM) ativados in vitro com estas duas espécies de bactérias. Demonstramos que IL-12p40 e TNF-a foram produzidos pelos MfDM ativados com qualquer uma das bactérias. A síntese de IL-12p40 foi dependente de TNF-a, MyD88 e NFkB e independente de MAPK p38 e ERK 1/2. Durante a estimulação com B. pertussis a produção de IL-12p40 foi dependente de TLR-4, mas com B. parapertussis envolveu outras vias independentes de MyD88 e TLR-4. Estas bactérias não induziram a síntese de IL-12p70, necessitando de sinais moleculares adicionais de IFN-g, que aumentou a síntese desta citocina. A produção de IL-12 p70 aumentou após o bloqueio das vias PI3K, MAPK p38 e ERK1/2 assim como após a adição exógena de PT sobre MfDM ativados com B. parapertussis. Portanto, diversas vias de sinais dependentes e independentes de TLR-4 controlam a produção de IL-12 neste modelo. / Bordetella pertussis and B. parapertussis are etiological agents of whooping cough. IL-12 links the innate and adaptive immunity. We investigated the ability of both bacteria to modulate IL-12 by in vitro activation of bone marrow derived macrophages (MfDM). We demonstrated that IL-12p40 and TNF-a were produced after stimulation of cells with either bacterium. IL-12p40 production was dependent on TNF-a, MyD88 and NFkB but independent of MAPK p38 and ERK 1/2. During B. pertussis activation the production of IL-12p40 was dependent on TLR-4, while B. parapertussis activation was MyD88 and TLR-4 independent. However, the bacteria alone did not induce IL-12p70 synthesis, requiring IFN-g as an additional signal. Evidences indicated MAPK p38, ERK1/2 and PI3K during B. pertussis and B. parapertussis activation, as well as the exogenous addition of PT to B. parapertussis activated MfDM, was critical for the up regulation of IL-12p70. This finding indicates that different TLR-4 dependent and independent signaling pathways may control the production of IL-12 in this model.

As vacinas pertussis celulares apresentam certa reatogenicidade e as acelulares, menos reatogênicas, têm seu uso restrito, devido a seu alto custo. O Instituto Butantan desenvolveu uma vacina pertussis celular (Plow), com baixo teor de lipopolissacarídeo e outra acelular (Pa), por metodologia simples e econômica. Essas preparações, combinadas aos toxóides diftérico e tetânico (DTPlow e DTPa, respectivamente), foram comparadas à DTP tradicional, com diferentes adjuvantes: vitamina A, surfactante pulmonar, BCG, monofosforil lipídeo A (MPL) e Al(OH)3. A resposta humoral em camundongos foi semelhante para as diferentes formulações e independente do adjuvante utilizado. As vacinas induziram níveis equilibrados de IgG1/IgG2a anti-pertussis e a DTPlow mostrou-se menos reatogênica, induzindo níveis significativamente menores de IL-6 sérica. A adição de MPL sugeriu tendência de proteção contra a colonização nasotraqueal no grupo imunizado com DTPa e levou à indução de IFN-g nos grupos imunizados com DTP e DTPa, sugerindo possível atividade imunomodulatória para Th1. / The whole cell pertussis vaccines present some reactogenicity and the acellular, less reactogenic, have prohibitive use due to its high cost. Instituto Butantan developed a less reactogenic whole cell pertussis vaccine (Plow), with low lipopolysaccharide content and an acellular vaccine (Pa), by simple and economic methodology. These preparations, combined to diphtheria and tetanus toxoids (DTPlow and DTPa, respectively), were compared with the traditional DTP, with different adjuvants: vitamin A, pulmonary surfactant, BCG, monophosphoryl lipid A (MPL) and Al(OH)3. The humoral immune response induced in mice by the different vaccine formulations, was similar and independent of the adjuvant used. The vaccines induced balanced levels of IgG1/IgG2a anti-pertussis and DTPlow showed to be less rectogenic, inducing lower levels of serum IL-6. The use of MPL suggested to induce higher protection against nasotracheal colonization in DTPa group and induced IFN-g in the DTP and DTPa groups, suggesting a possible immunemodulatory activity for Th1.

La coqueluche est une pathologie due à la bactérie Bordetella pertussis qui touche les voies respiratoires des patients infectés causant toux, leucocytose, fièvre, et dont les symptômes peuvent aller jusqu’au décès chez les individus les plus à risque (nouveau-nés et enfants immunodéprimés en particulier). Ciblée par différents programmes vaccinaux depuis de nombreuses années, cette pathologie sévit à nouveau dans de nombreux pays développés où le nombre de cas augmente fortement depuis la fin des années 2000. Cette résurgence montre la nécessité de développer de nouvelles stratégies afin de comprendre les mécanismes de l’infection par B. pertussis. Dans ce contexte, la recherche préclinique apparaît comme essentielle pour comprendre la physiopathologie de la coqueluche. De nombreux modèles animaux ont été décrits pour l’étude de la coqueluche mais aucun de ces modèles ne permet de reproduire l’ensemble du spectre des symptômes cliniques de la pathologie, notamment la toux. Cependant, au cours des dernières années un modèle d’infection par Bordetella pertussis chez le jeune babouin a été développé aux Etats-Unis et permet de reproduire la pathologie observée chez l’homme, notamment concernant la toux et la transmission. Ce modèle semble ainsi très prometteur pour l’étude de la physiopathologie de la coqueluche.Cependant, de nombreuses inconnues subsistent dans ce modèle, notamment concernant la colonisation bactérienne et les interactions entre la bactérie et l’hôte. Nous avons ainsi cherché dans cette étude à évaluer d’une part l’impact de différents facteurs comme l’âge des animaux, la dose d’infection ainsi que la voie d’exposition sur la pathologie déclarée par les babouins suite à l’infection par la souche B1917 de B. pertussis afin de pouvoir proposer un parallèle avec les données cliniques disponibles. Nous avons également développé l’utilisation de techniques d’imagerie in vivo comme l’endomicroscopie confocale couplée à la bronchoscopie afin d’étudier la localisation et la cinétique de colonisation et certaines interactions du pathogène dans le tractus respiratoire inférieur au cours de la pathologie. Cette étude nous a ainsi permis d’approfondir les connaissances de physiopathologie de la coqueluche dans ce modèle babouin et consolidera cet outil précieux pour l’évaluation des futures stratégies de prévention contre cette pathologie. / Whooping cough, or pertussis, is a respiratory disease caused by Bordetella pertussis bacterial colonization of human airways. Main symptoms are cough, leukocytosis, fever and may even be lethal for some patients (e.g. newborn infants and immuno-deficient patients). Despite a good vaccination coverage worldwide against pertussis, whooping cough cases have been re-increasing in several developed countries in the past twenty years. This resurgence points out the crucial need to develop new control strategies and to better understand pertussis pathophysiology, notably using appropriate animal models. Numerous preclinical models including mice, rats, rabbits and swine have been described for B. pertussis infection studies. However, none of these models reproduce the full spectrum of clinical pertussis symptoms, especially cough. The recent baboon model of whooping cough described in the last few years in the US appears to be a very relevant model for pertussis pathophysiology studies as these animals reproduced all clinical symptoms as observed in humans including cough.However, many aspects of bacterial colonization and interactions with the host have yet to be described in this model.We have then evaluated diverse parameters such as animal age, the inoculum dose and the exposition route on the pathology symptoms and immune responses developed by baboons following B. pertussis B1917 strain inoculation in order to draw a parallel with human clinical data. We also developed in this model in vivo imaging techniques like confocal endomicroscopy coupled with bronchoscopy in order to evaluate bacterial colonization kinetics, localization and some interactions in the lower respiratory tract of infected baboons. Then, this study brought additional data on whooping cough physiopathology in this baboon model, which will be crucial for evaluating future prevention strategies against pertussis disease

<p>Uprkos rezultatima postignutih imunizacijom, veliki ka&scaron;alj je i danas vodeći uzrok smrti među zaraznim bolestima protiv kojih se sprovodi imunizacija. Postojanje različitih vrsta nadzora, prisutnih nedostataka laboratorijske dijagnostike oboljenja, slabosti u dostupnim definicijama slučaja velikog ka&scaron;lja i neprepoznavanje oboljenja, otežavaju realno sagledavanje opterećenja velikim ka&scaron;ljem i poređenje postignutih rezultata u različitim zemljama. Postojeće definicije slučaja velikog ka&scaron;lja nisu prihvatljive u svim uzrastima. Cilj istraživanja je bio da se na reprezentativnom uzorku Grada Novog Sada utvrdi: stopa incidencije velikog ka&scaron;lja tokom jedne godine savremenog nadzora; senzitivnost i specifičnost definicija slučaja velikog ka&scaron;lja, datih od strane Globalne pertusis inicijative za tri uzrasne grupe. U istraživanje je uključeno 213 ispitanika pacijenata iz sentinelnog nadzora nad velikim ka&scaron;ljem Doma zdravlja Novi Sad i 107 ispitanika pacijenata iz hospitalnog nadzora sa iste teritorije koji su ispunjavali predložene definicije slučaja za tri uzrasne grupe (0-3 meseca; 4 meseca-9 godina života; 10 godina i stariji). Podaci za istraživanje dobijeni su popunjavanjem anketnog upitnika. Laboratorijsko testiranje sumnji na veliki ka&scaron;alj sprovedeno je u Centru za mikrobiologiju Instituta za javno zdravlje Vojvodine. Kod pacijenata sa ka&scaron;ljem u trajanju do 21 dan testiranje je vr&scaron;eno upotrebom PCR metoda, a kod pacijenata sa ka&scaron;ljem u trajanju vi&scaron;e od 21 dan, dokazivanje oboljenja vr&scaron;eno je upotrebom ELISA IgA i IgG serolo&scaron;kih testova. Potvrđenim slučajem velikog ka&scaron;lja smatran je pacijent sa ispunjenom definicijom slučaja oboljenja i laboratorijskom potvrdom oboljenja, PCR ili ELISA testom. Procenjena stopa incidencije velikog ka&scaron;lja u Novom Sadu je 237,7/100.000, a stopa incidencije hospitalizovanih u Novom Sadu je 16,4/100.000. Najvi&scaron;e stope incidencije velikog ka&scaron;lja u sentinelnom nadzoru registruju se u uzrastu 10-14, a u hospitalnom u uzrastu od 7 do 9 godina. Senzitivnost, specifičnost i stepen verovatnoće pozitivnog rezultata testiranog simptoma/znaka iz predloženih definicija slučaja se razlikuju po uzrastima i po pojedinim simptomima/znacima. Predložene definicije slučaja u uzrastima od 4 meseca do 9 godina i u uzrastu od 10 godina i starijih imaju veću verovatnoću otkrivanja obolelih u hospitalnom u odnosu na sentinelni nadzor za pojedine simptome/znakove. Budući da je tokom istraživanja u sentinelnom nadzoru oboljenje potvrđeno kod svakog petog, a u hospitalnom kod svakog drugog testiranog pacijenta, predložene definicije slučaja se mogu koristiti u nadzoru nad velikim ka&scaron;ljem.</p> / <p>Despite all results achieved by immunization, pertussis is still the leading cause of death among vaccine preventable diseases. Different types of surveillance and laboratory confirmation of pertussis, weakness of existing case definitions for pertussis and broad spectrum clinical manifestation of disease, complicate overview of disease and result comparison of surveillance in different countries. The current pertussis case definition is not acceptable for all age groups of patients. The aim of this research was to determine: the pertussis incidence rate in population of Novi Sad during one year of modern surveillance; the sensitivity and specificity of clinical case definition for pertussis, given by the Global Pertussis Initiative for three age groups, with a representative sample of population in the City of Novi Sad. 213 patients from sentinel surveillance of pertussis Novi Sad Health Centre and 107 hospitalized patients from Novi Sad, who fulfilled criteria of case definition proposed for the three age groups (0-3 months, 4 months-9 years; 10 years of age and older), were included in the research. Research data obtained from a questionnaire. Laboratory testing of suspected cases were conducted at the Centre for Microbiology, Institute of Public Health of Vojvodina. For patients with coughing less than 21 days PCR method was used, and for patients with cough lasting more than 21 days, laboratory confirmation of disease was performed using ELISA IgA and IgG serological tests. Confirmed case of pertussis consider to be a patient with symptoms/signs according to proposed case definition and with laboratory confirmation of the pertussis, PCR or ELISA. Estimated incidence rate of pertussis for population in the city of Novi Sad was 237.7/100,000, and the pertussis incidence rate in hospitalized patient in Novi Sad was 16.4/100,000. The highest incidence rate of pertussis in the sentinel surveillance was registered in the age group 10-14 and in the hospital surveillance in the group 7 to 9 years of age. The values of sensitivity, specificity and positive likelihood ratio of symptoms/signs from the proposed case definition were calculated and they different by age and by certain symptoms/signs from proposed case definition. Certain symptoms/signs of the proposed case definition have a higher probability of detection among patients aged 4 months to nine, and at the age of ten year and older, in the hospital versus those in sentinel surveillance for pertussis. During research from the patients who were tested in sentinel surveillance every fifth was laboratory confirmed case and in the hospital surveillance every second patient was confirmed case, so the proposed case definitions can be used in the surveillance of pertussis.</p>

Bordetella pertussis is a strictly human pathogen colonizing the upper respiratory tract, causing a respiratory disease known as whooping cough or pertussis. The introduction of whole-cell vaccines and acellular vaccines, resulted in a significant reduction in the incidence of disease and reduce the fatalities associated with infection. However, epidemiological data show a significant increase in the incidence of the disease in recent decades. The increasing incidence is mainly attributed to the transition from the whole- cell vaccine to an acellular vaccine. Based on research from recent years has shown that acellular vaccines have many drawbacks, and it is therefore necessary to change the vaccination strategy. One possible solution to the situation is the development of a new generation of whole-cell vaccines with reduced reactogenicity. The new whole-cell vaccine was prepared by a genetically modified B. pertussis strain. B. pertussis was modified using allelic exchange to develop strain encoding enzymatically inactive pertussis toxin, modified lipid A and lacking dermonecrotic toxin. This combination of genetic modifications in mice led to a decrease in reactogenicity test vaccine in vivo. In case of intranasal infection whole-cell vaccine containing genetically modified strain is providing...