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Perfil imunoproteômico da resposta humoral na revacinação com Mycobacterium bovis BCG Moreau, a cepa vacinal brasileira contra tuberculosePereira, Melissa Pontes January 2009 (has links)
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Previous issue date: 2009 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / A tuberculose (TB) ocupa o segundo lugar em causa de morte por doença infecciosa de notificação compulsória. Uma das estratégias para seu controle é a vacinação, e a única vacina disponível é o BCG (Bacilo de Calmette-Guèrin). Embora utilizada mundialmente, a vacina BCG apresenta diversas limitações. O Brasil utiliza a cepa Moreau para produção da vacina, sendo o único país do mundo autilizar, desde os anos 1930, esta cepa para vacinação. Como observado para outros patógenos intracelulares, a imunidade celular desempenha papel importante na proteção do hospedeiro contra a TB. O IFNy, secretado por células T, é descrito como uma citocina particularmente importante participando na resposta imune protetora, e é empregado como correlato de proteção, embora sua utilidade seja ainda questionável, sobretudo em populações adultas. Isto evidencia a necessidade de buscar biomarcadores adicionais que permitam uma correlação de proteção mais confiável. As proteínas extracelulares micobacterianas estão entre as primeiras moléculas a interagirem com o sistema imune do hospedeiro, e uma melhor compreensão de seu papel imunomodulatório poderá contribuir para o desenvolvimento de novos alvos para o controle e erradicação da TB. Uma estratégia para a identificação de proteínas imunologicamente relevantes é a análise do imunoproteoma, que combina a abordagem proteômica com uma apresentação imunológica dos dados. A caracterização de soros, provenientes de indivíduos vacinados, muitas vezes revela diferentes padrões de reconhecimento que podem ser indicativos dos diferentes graus de proteção conferidos pela vacina em uma população heterogênea
Empregamos assim esta tecnologia, combinando a separação por eletroforese bi-dimensional (2DE) de antígenos protéicos extracelulares da cepa vacinal brasileira seguida de western-blot, para avaliar o perfil imunoproteômico da resposta humoral de indivíduos saudáveis, PPD negativos, revacinados com BVG Moreau e classificados como alto e baixo produtores de IFNy. Nossos resultados revelam um padrão de reconhecimento claramente distinto entre os grupos avaliados, correlacionado com o perfil de produção de IFNy. Dentre as proteínas imunoreativas identificadas, um grupo (antígenos do complexo 85) foi reconhecido por todos os soros incluídos no estudo, inclusive controles. Alguns antígenos foram reconhecidos exclusivamente pelos indivíduos classificados como alto produtores de IFNy e outros somente pelos indivíduos enquadrados no grupo de baixo produtores. Algumas proteínas foram selecionadas e produzidas sob a forma recombinante em Escherichia coli possibilitando sua avaliação mais detalhada frente aos soros individualizados. Entre os antígenos diferencialmente reconhecidos, as proteínas Mpb70 e Mpb83 se destacaram como potenciais correlatos da proteção oferecida pela vacinação com BCG Moreau, tendo sido reconhecidos somente pelos indivíduos produtores de elevados níveis de IFNy. Estudos adicionais são necessários para determinar a aplicabilidade efetiva de tais antígenos como potenciais correlatos da proteção conferida pela vacinação com BCG Moreau. Este é o primeiro relato que detalha o perfil de reconhecimento sorológico por indivíduos revacinados com BCG Moreau frente a antígenos extracelulares da cepa homóloga, e contribui para uma melhor compreensão da cepa vacinal brasileira contra a TB / Tuberculosis (TB) holds the second place as cause o
f death by an infectious
disease of obligatory notification. One of the stra
tegies for its control is vaccination,
and the only vaccine available is BCG (Bacillus Cal
mette-Guerin). Although used
worldwide, BCG shows various limitations. Brazil us
es the Moreau strain for vaccine
production, being the only country in the world to
use, since the 1930s, this strain for
vaccination. As observed for other intracellular pa
thogens, cellular immunity plays an
important role in host protection against TB. IFN
γ
secreted by T cells is described as
a particularly important cytokine participating in
protective immune responses, being
used as a correlate of protection, although its uti
lity remains questionable, especially
in adult populations. This highlights the need to s
eek additional biomarkers yielding
better surrogates of protection. Mycobacterial extr
acellular proteins are among the
first molecules to interact with the host immune sy
stem and a better understanding of
their immunomodulatory role may contribute to the d
evelopment of new targets for
the control and eradication of TB. A strategy for t
he identification of immunologically
relevant proteins is the analysis of the immunoprot
eome, combining a proteomic
approach with an immunological presentation of the
data. The characterization of
sera from vaccinated individuals often reveals diff
erent patterns of recognition that
may be indicative of the different degrees of prote
ction conferred by the vaccine in a
heterogeneous population. We have thus employed thi
s technology, combining the
separation by two-dimensional electrophoresis (2DE)
of extracellular proteins from
the Brazilian vaccine strain, followed by Western b
lot to assess the
immunoproteomic profile of the humoral response in
healthy, PPD negative subjects,
re-vaccinated with BCG Moreau and classified as hig
h or low producers of IFN
γ
. Our
results reveal a clearly distinct recognition patte
rn between the evaluated groups,
correlating with the profile of IFN
γ
production. Among the immunoreactive proteins
identified, one group (85 complex antigens) was rec
ognized by all sera included in
the study, including controls. Some antigens were r
ecognized only by individuals
classified as high producers of IFN
γ
and others only by the individuals included in the
group of low producers. Some proteins were selected
and produced in recombinant
form in
Escherichia coli
allowing for their further evaluation against the
individual
sera. Among the antigens differentially recognized,
Mpb70 and Mpb83 proteins stood
out as potential correlates of protection offered b
y vaccination with BCG Moreau, and vi
were only recognized by individuals producing high
levels of IFN
γ
. Additional studies
are needed to determine the effective applicability
of these antigens as potential
correlates of protection provided by vaccination wi
th BCG Moreau. This is the first
report detailing the serologic recognition profile
of individuals re-vaccinated with BCG
Moreau against extracellular antigens of the homolo
gous strain, and contributes to a
better understanding of the Brazilian vaccine strai
n against TB.
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Fonctions nonconvexes inférieurement s-régulières / Nonconvex s-lower regular functionsKecis, Ilyas 17 June 2014 (has links)
La thèse est constituée de cinq chapitres. Le premier chapitre est consacré à étudier le sous-différentiel de Hölder ainsi que le cône normal de Hölder. Nous établissons différentes règles de calcul pour ce type de sous-différentiel. La relation entre le sous-différentiel de Hölder de la fonction distance et le cône normal de Hölder d'un ensemble S en un point x est également traitée dans le cas où x est dans S ou en dehors de S. Le deuxième chapitre étudie les fonctions inférieurement $s$-régulières dans un espace de Banach. Cette classe de fonctions est une extension de celle dite "Primal lower nice functions" (pln en abrégé), introduite par R.A. Poliquin dans les espaces de dimension finie. Le but de cette partie est de donner dans le contexte d'espaces Banachiques plus généraux, une caractérisation sous-différentielle de ces fonctions ainsi que l'égalité avec d'autres sous-différentiels connus. Nous nous intéressons dans le troisième chapitre à l'étude des propriétés de différentiabilité de l'enveloppe de Moreau d'une fonction inférieurement $s$-régulière. Nous établissons entre autres, sous des conditions assez générales, que l'enveloppe de Moreau d'une telle fonction est de classe C^{1,alpha} et que l'application proximale associée est Höldérienne.Dans les chapitres 4 et 5, nous obtenons des résultats d'existence de solutions d'inégalités variationnelles. Nous considérons le cas d'inclusion différentielle associée au sous-différentiel d'une fonction plr avec une perturbation. Le cas d'inclusion différentielle gouvernée par le cône normal d'un ensemble prox-régulier est aussi étudié. / The thesis contains five chapters. The first chapter is devoted to study the Hölder subdifferential and the Hölder normal cone. We establish different calculus rules for this type of subdifferential. The relationship between the Hölder subdifferential of the distance function and the Hölder normal cone of a set S at a point x is also studied in the case where either x is in S or outside of S. The second chapter studies the s-lower regular functions in a Banach space. This class of functions is an extension of the Primal lower nice functions ( pln for short) introduced by R.A. Poliquin in finite dimensional spaces. The purpose of this section is to establish in the context of general Banach spaces, a subdifferential characterization of these functions as well as the equality with other known subdifferentials. We are interested in the third chapter in the study of differentiability properties of the Moreau envelope associated to an s-lower regular function. We show, under enough general conditions that, the Moreau envelope of such functions is of class C^{1,alpha} and the associated proximal mapping is Hölderian. In chapters 4 and 5, we obtain existence results of solution of variational inequalities. We consider the case of differential inclusion associated to plr functions with single-valued perturbation. The case of differential inclusion governed by the normal cone of prox-regular sets is also studied.
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A stylistic analysis of selected piano works of Louis Moreau GottschalkDicus, Kent Timothy, 1958- January 1988 (has links)
Although Louis Moreau Gottschalk's works are not generally recognized as being exceptionally significant in the development of musical style, they do serve as a link between the music of Frederic Chopin and that of Charles Ives. Certain stylistic characteristics of Chopin are seen in many of Gottschalk's works, especially those which incorporate "Scherzo" and "Mazurka" passages. Simultaneously, Gottschalk's concept of using popular tunes as prominent melodies and themes was later expanded by Charles Ives. Gottschalk's works include some of America's first experimentations with form through utilization and expansion of the basic form of ABA Coda. Through his use of varied ABA form with repeated and parallel passages, Gottschalk developed his particular style of phrasing, texture, and rhythm, all of which figure prominently in his works. Four pieces are examined with these concepts as the basis for analysis.
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Barrier spit evolution and primary consolidation of backbarrier facies: West Belle Pass Barrier, LAKramer, John N, III 13 May 2016 (has links)
West Belle Pass Barrier is a barrier spit that formed during the last delta lobe progradation associated with the Lafourche delta complex. Located on the western flank of the Caminada-Moreau Headland, West Belle Pass Barrier and Raccoon Pass are located downdrift of the Belle Pass jetties. Morphological changes stemming from storms, jetty infrastructure, and an expanding tidal inlet are evaluated using historical shoreline data and imagery. Littoral transport around the jetties combined with inlet growth created a framework wherein sediment is transported through Raccoon Pass and sequestered as a flood-tidal delta. These events aided in the landward migration of West Belle Pass Barrier, which ultimately loaded and consolidated previously unconsolidated facies. A conceptual model illustrating the primary consolidation of backbarrier facies resulting from washover deposits during one storm is presented. The primary consolidation settlement associated with loading near-surface, water-saturated backbarrier facies is substantially larger than current subsidence rates.
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Gilbert, Malfilatre, Hégésippe Moreau et quelques autres poètes mauditsDe La Mata, Claudine. January 1986 (has links)
Th. 3e cycle--Litt. fr.--Toulouse 2, 1985.
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Littérature et drogues au XIXe siècle. De Rabbe à Baudelaire / Literature and Drugs in the 19th Century France. From Rabbe to BaudelaireYogo, Takenori 10 December 2010 (has links)
Homère, Hérodote et Pline avaient déjà décrit les effets merveilleux de la drogue. Abordé par Rabelais, Mme de Sévigné et Chardin, le motif du stupéfiant dans la littérature a connu un âge d’or dès la première moitié du XIXe siècle. Poussés par une aspiration à fuir vers l’infini, sinon par une simple curiosité pour un univers inconnu, les écrivains romantiques se mettent à consommer de l’opium et du haschisch, et commencent à les introduire dans leurs œuvres comme motif littéraire dans des circonstances historiques et culturelles. La présente thèse analyse l’évolution de l’emploi littéraire de la drogue qui implique divers contextes, à savoir médical, social et anthropologique, au travers des textes de Rabbe, Musset, Balzac, Gautier, Dumas, Nerval et Baudelaire, et de certains phénomènes culturels tels que la légende de l’Ordre des Assassins et le club des haschischins à l’hôtel Pimodan. / Broached by Rabelais, Madame de Sévigné and Chardin, the motif of narcotics in literature enjoyed a golden age as early as the first half of the 19th Century, although Homer, Herodotus and Pliny had already described their wonderful effects. Driven by an aspiration to escape to the infinite, or by simple curiosity for an unknown universe, the Romantics started to consume opium and hashish, and started to introduce them into their work as a literary motif in historic and cultural circumstances. This thesis analyses the evolution of the use of drugs in literature, in a wide range of contexts, namely medical, social and anthropological, through the texts of Rabbe, Musset, Balzac, Gautier, Dumas, Nerval, and Baudelaire, and certain cultural phenomena such as the legend of the Order of Assassins and the club des Haschischins at hôtel Pimodan.
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Louis Moreau Gottschalk (1829-1869): The Role of Early Exposure to African-Derived Musics in Shaping an American Musical Pioneer From New OrleansUnruh, Amy Elizabeth 12 November 2009 (has links)
No description available.
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Some aspects on sweeping processes / Quelques résultats sur les processus de rafleLatreche, Wissam 10 July 2018 (has links)
Dans cette thèse, on s'intéresse à l'étude d'existence de solutions pour les processus de rafle. Ce problème prend la forme d'une inclusion différentielle contrainte avec des cônes normaux qui apparaissent naturellement dans nombreuses applications telles que le mouvement de foule, l'élastoplasticité, les mécaniques, les circuits électroniques, etc. L'objective de ce travail est de rapprocher deux importantes classes d'inclusions différentielles. D'une part, nous établissons quelques résultats d'existence de tube-solutions pour des processus de rafle à des ensembles uniformément prox-réguliers. D'autre part, nous présentons des résultats d'existence de solutions monotone par rapport à un préordre pour un système mixte d'inclusions différentielles projetées. De plus, nous montrons l'existence d'un point-selle pour notre système et nous fournissons deux exemples d'applications. / In this thesis, we were interested in the study of the existence of solutions for sweeping processes. This problem takes the form of a constrained differential inclusion involving normal cones which appears naturally in many applications such as crowd motion, elastoplasticity, mechanics, electrical circuit, etc.The aim of this work is to bring together two classes of differential inclusions. On one hand, we establish some existence results of solutions-tube for sweeping processes with uniformly prox-regular sets. On the other hand, we present existence results of monotone solutions with respect to a preorder for a mixed system of projected differential inclusions. In addition, we show that our system has a saddle-point and we provide two examples of applications.
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Construção, clonagem e expressão do fragmento B da toxina diftérica de Corynebacterium diphtheriae (cepa PW- 8) em Mycobacterium bovis BCG sub-cepa Moreau / Construction, cloning and expression of the fragment B of diphtheria toxin from Corynebacterium diphtheriae (strain PW-8) in Mycobacterium bovis BCG Moreau sub-strainDilzamar Veloso do Nascimento 28 March 2014 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A vacina anti-diftérica de uso corrente no Brasil (DTP), embora de alta eficácia na prevenção da difteria, está associada com episódios de toxicidade e reatogenicidade no recipiente vacinal, resultantes de proteínas residuais derivadas do processo de produção ou detoxificação. Estratégias para o desenvolvimento de vacinas menos reatogênicas e ao mesmo tempo mais eficazes e economicamente viáveis contra a difteria têm sido alvo de intensa investigação. A alternativa proposta por nosso grupo é a utilização da vacina contra a tuberculose (Mycobacterium bovis BCG sub-cepa Moreau), como vetor do gene que codifica o fragmento B da toxina diftérica (dtb) de 58,3 kDa. Neste trabalho o dtb foi clonado no vetor micobacteriano bifuncional (pUS977) de expressão citoplasmática e os clones recombinantes (pUS977dtbPW8), após a transformação do BCG, foram testados com relação a expressão do DTB em BCG e quanto a antigenicidade frente a anticorpos policlonais anti-toxóide diftérico por Immunobloting. A integridade do gene dtb e a identidade das sequências de DNA da construção plasmidial pUS977dtbPW8 foram confirmadas por sequenciamento de DNA e análise de similaridade. A imunogenicidade do BCGr pUS977dtbPW8 expressando o DTB foi investigada em camundongos BALB/c, os resultados obtidos revelaram uma soroconversão específica (IgG). A infectividade e atividade microbicida do BCGr pUS977dtbPW8 no ambiente intracelular foi avaliada através da infecção de linhagens de células de monócitos humano (THP-1), os dados obtidos indicaram que houve sobrevivência intracelular em até 12 dias. Nesse contexto, esplenócitos dos camundongos imunizados com 30 e 60 dias foram extraídos, mostrando que o BCGr pUS977dtbPW8 persistiu até 60 dias na ausência de pressão seletiva e a viabilidade celular não sofreu alteração significativa durante o período testado. Por outro lado, o BCGr pUS977dtbPW8, quando submetido a seis sub-cultivos consecutivos in vitro não apresentou diferença significativa na capacidade de expressar o DTB, demonstrando portanto a persistência da estabilidade funcional da linhagem recombinante. A estabilidade estrutural da construção pUS977dtbPW8 também foi avaliada por PCR confirmando a presença do gene dtb em colônias do BCGr pUS977dtbPW8 . Adicionalmente, foi possível avaliar preliminarmente in vitro a capacidade soroneutralizante dos soros de camundongos imunizados com BCGr pUS977dtbPW8 após 30 e 60 dias em células VERO. A ação citotóxica da toxina diftérica entre as diluições de 1/4 e 1/16 foram neutralizadas com o pool de soros imunes com 60 dias. Finalmente, em nosso estudo foi possível avaliar o potencial da vacina BCG como vetor de expressão de um antígeno de Corynebacterium diphtheriae in vitro e in vivo. / The diphtheria vaccine currently used in Brazil (DTP), despite its history of high efficacy in the prevention of diphtheria, is associated with episodes of toxicity and vaccine reactogenicity in the vaccinee, resulting from the presence in the vaccine of residual proteins derived from the production process or detoxification. Strategies for the development of new vaccines more effective and economically viable against diphtheria have been the subject of intense investigation. The alternative proposed by our group is the use of the vaccine against tuberculosis (Mycobacterium bovis BCG Moreau sub strain) as a vector for the gene that encodes the 58.3 kDa fragment B of the diphtheria toxin (DTB). In our project the dtb gene was cloned into the bifunctional vector pUS977 for cytoplasmic expression and recombinant BCG (rBCG) clones, selected after transformation of BCG, were tested for expression of the DTB polypeptide and antigenicity against polyclonal antibodies anti- diphtheria toxoid by immunoblotting. The integrity and identity of the DNA sequence encoding the dtb gene carried by the plasmid construct pUS977dtbPW8 was confirmed by DNA Sequencing and Analysis of Similarity. The immunogenicity of the rBCG expressing the DTB was investigated in BALB/c mice and the results revealed a specific seroconversion (IgG). Also, infectivity and microbicidal activity were analyzed in the intracellular environment by infecting human monocytes (THP-1 cell line) with rBCG. The data obtained indicated intracellular survival within 12 days. In this context, splenocytes collected from mice at days 30 and 60 after immunization were removed and assayed for live bacteria. The results showed that rBCG persisted viable up to 60 days in the absence of selective pressure and cell viable counts did not change significantly during testing. Additionally, the rBCG subjected to six consecutive sub-cultures in vitro showed no significant difference in the ability to express the DTB, thus demonstrating the functional stability of the recombinant vaccine. The structural stability of the construct pUS977dtbPW8 was also confirmed by PCR detection of the dtb gene in rBCG colonies. Also, it was possible to have a preliminary evaluation of the neutralizing capacity of sera from mice immunized with BCGr 30 and 60 days after immunization. The cytotoxic action of diphtheria toxin, between dilutions 1/ 4 and 1/16, was neutralized by mice sera in an in vitro assay using VERO cells. Finally, in our study it was possible to evaluate the potential of BCG as a vector for expression of an antigen of Corynebacterium diphtheriae in vitro and in vivo.
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Construção, clonagem e expressão do fragmento B da toxina diftérica de Corynebacterium diphtheriae (cepa PW- 8) em Mycobacterium bovis BCG sub-cepa Moreau / Construction, cloning and expression of the fragment B of diphtheria toxin from Corynebacterium diphtheriae (strain PW-8) in Mycobacterium bovis BCG Moreau sub-strainDilzamar Veloso do Nascimento 28 March 2014 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A vacina anti-diftérica de uso corrente no Brasil (DTP), embora de alta eficácia na prevenção da difteria, está associada com episódios de toxicidade e reatogenicidade no recipiente vacinal, resultantes de proteínas residuais derivadas do processo de produção ou detoxificação. Estratégias para o desenvolvimento de vacinas menos reatogênicas e ao mesmo tempo mais eficazes e economicamente viáveis contra a difteria têm sido alvo de intensa investigação. A alternativa proposta por nosso grupo é a utilização da vacina contra a tuberculose (Mycobacterium bovis BCG sub-cepa Moreau), como vetor do gene que codifica o fragmento B da toxina diftérica (dtb) de 58,3 kDa. Neste trabalho o dtb foi clonado no vetor micobacteriano bifuncional (pUS977) de expressão citoplasmática e os clones recombinantes (pUS977dtbPW8), após a transformação do BCG, foram testados com relação a expressão do DTB em BCG e quanto a antigenicidade frente a anticorpos policlonais anti-toxóide diftérico por Immunobloting. A integridade do gene dtb e a identidade das sequências de DNA da construção plasmidial pUS977dtbPW8 foram confirmadas por sequenciamento de DNA e análise de similaridade. A imunogenicidade do BCGr pUS977dtbPW8 expressando o DTB foi investigada em camundongos BALB/c, os resultados obtidos revelaram uma soroconversão específica (IgG). A infectividade e atividade microbicida do BCGr pUS977dtbPW8 no ambiente intracelular foi avaliada através da infecção de linhagens de células de monócitos humano (THP-1), os dados obtidos indicaram que houve sobrevivência intracelular em até 12 dias. Nesse contexto, esplenócitos dos camundongos imunizados com 30 e 60 dias foram extraídos, mostrando que o BCGr pUS977dtbPW8 persistiu até 60 dias na ausência de pressão seletiva e a viabilidade celular não sofreu alteração significativa durante o período testado. Por outro lado, o BCGr pUS977dtbPW8, quando submetido a seis sub-cultivos consecutivos in vitro não apresentou diferença significativa na capacidade de expressar o DTB, demonstrando portanto a persistência da estabilidade funcional da linhagem recombinante. A estabilidade estrutural da construção pUS977dtbPW8 também foi avaliada por PCR confirmando a presença do gene dtb em colônias do BCGr pUS977dtbPW8 . Adicionalmente, foi possível avaliar preliminarmente in vitro a capacidade soroneutralizante dos soros de camundongos imunizados com BCGr pUS977dtbPW8 após 30 e 60 dias em células VERO. A ação citotóxica da toxina diftérica entre as diluições de 1/4 e 1/16 foram neutralizadas com o pool de soros imunes com 60 dias. Finalmente, em nosso estudo foi possível avaliar o potencial da vacina BCG como vetor de expressão de um antígeno de Corynebacterium diphtheriae in vitro e in vivo. / The diphtheria vaccine currently used in Brazil (DTP), despite its history of high efficacy in the prevention of diphtheria, is associated with episodes of toxicity and vaccine reactogenicity in the vaccinee, resulting from the presence in the vaccine of residual proteins derived from the production process or detoxification. Strategies for the development of new vaccines more effective and economically viable against diphtheria have been the subject of intense investigation. The alternative proposed by our group is the use of the vaccine against tuberculosis (Mycobacterium bovis BCG Moreau sub strain) as a vector for the gene that encodes the 58.3 kDa fragment B of the diphtheria toxin (DTB). In our project the dtb gene was cloned into the bifunctional vector pUS977 for cytoplasmic expression and recombinant BCG (rBCG) clones, selected after transformation of BCG, were tested for expression of the DTB polypeptide and antigenicity against polyclonal antibodies anti- diphtheria toxoid by immunoblotting. The integrity and identity of the DNA sequence encoding the dtb gene carried by the plasmid construct pUS977dtbPW8 was confirmed by DNA Sequencing and Analysis of Similarity. The immunogenicity of the rBCG expressing the DTB was investigated in BALB/c mice and the results revealed a specific seroconversion (IgG). Also, infectivity and microbicidal activity were analyzed in the intracellular environment by infecting human monocytes (THP-1 cell line) with rBCG. The data obtained indicated intracellular survival within 12 days. In this context, splenocytes collected from mice at days 30 and 60 after immunization were removed and assayed for live bacteria. The results showed that rBCG persisted viable up to 60 days in the absence of selective pressure and cell viable counts did not change significantly during testing. Additionally, the rBCG subjected to six consecutive sub-cultures in vitro showed no significant difference in the ability to express the DTB, thus demonstrating the functional stability of the recombinant vaccine. The structural stability of the construct pUS977dtbPW8 was also confirmed by PCR detection of the dtb gene in rBCG colonies. Also, it was possible to have a preliminary evaluation of the neutralizing capacity of sera from mice immunized with BCGr 30 and 60 days after immunization. The cytotoxic action of diphtheria toxin, between dilutions 1/ 4 and 1/16, was neutralized by mice sera in an in vitro assay using VERO cells. Finally, in our study it was possible to evaluate the potential of BCG as a vector for expression of an antigen of Corynebacterium diphtheriae in vitro and in vivo.
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