published_or_final_version / Biological Sciences / Master / Master of Philosophy

L’objectif de cette thèse est de formuler un adjuvant phytosanitaire qui permet d’optimiser les propriétés rhéologiques d’une préparation phytosanitaire (bouillie) afin de limiter la formation de gouttes susceptibles à la dérive lors de la pulvérisation, et d’améliorer le contact des gouttes avec le végétal traite. Les propriétés rhéologiques peuvent être optimisées par la présence de polysaccharides judicieusement sélectionnés. Comme la dissolution des polysaccharides représente une difficulté car ils forment des grumeaux, le passage par une émulsion permettant de les piéger dans la phase dispersée est nécessaire. La formulation est donc une émulsion de type eau-dans-huile (E/H) dont la phase dispersée est une solution concentrée de polysaccharide. La solution de polysaccharide est destinée par la suite a être relarguée lors de la préparation phytosanitaire dans un tank de pulvérisation. La formulation doit posséder une stabilité dans le temps répondant aux réglementations en vigueur pour les formulations phytosanitaires liquides. De ce fait, lastabilité de l’émulsion a été optimisée en utilisant la composition adéquate (phase continue, phase dispersée, tensioactif) et un procédé de fabrication optimise utilisant un système rotor-stator et un homogénéisateur à haute pression. Les formulations établies ont été caractérisées par des techniques permettant d’apprécier la taille moyenne et la distributionde la taille des gouttes dispersées telles que la granulométrie par diffraction laser et la calorimétrie a balayage différentielle (DSC). La température de cristallisation des gouttelettes mesurée lors de l’analyse DSC est corrélée à leur taille. La stabilité dans le temps a été évaluée en réalisant des tests de vieillissement naturel et des tests de vieillissement accéléré. Le relargage de polysaccharide a ensuite été étudiée afin de pouvoir être réalisé dans les conditions de préparation d’une bouillie au champ. Le relargage est réalisé par une déstabilisation préalable de la formulation à l’aide d’un tensioactif desémulsionnant. Le relargage est évalué par l’évolution de la conductivité électrique de la bouillie car les polysaccharides relargués s’ionisent en milieu aqueux. L’étude de la pulvérisation a par la suite été réalisée en utilisant un pilote reproduisant une rampe de pulvérisation agricole. La pulvérisation a été optimisée en faisant varier les propriétés physicochimiques de la préparation à pulvériser (tension de surface et rhéologie) et les paramètres de la pulvérisation (pression, buse de pulvérisation). La formulation de l’adjuvant retenue est une émulsion contenant 50 % (v/v) de solution de carboxymethylcellulose a 3 % (m/m), dispersée dans une phase huileuse constituée d’ester méthylique de colza avec 10 % (m/m) de polyricinoleate de polyglycérol. Le relargage est réalisé à l’aide d’un tensioactif éthoxylé, sélectionné parmi sept tensioactifs commerciaux. L’étude rhéologique de la formulation à pulveriser montre des propriétés qui permettent delimiter la formation des gouttes de petits diamètres, confirme par les essais de pulvérisation réalisés sur le pilote. / The objective of this thesis is to develop the formulation of a phytosanitary adjuvant to optimize the rheological properties of a preparation devoted to plant protection, in order to reduce the formation of droplets that may drift during spraying, and improve the contact of the droplets with the treated plant. Rheological properties can be optimized by the presence of wisely selected polysaccharides. As the dissolution of the polysaccharide is a problem because they form lumps, passing through the emulsion form to trap them in the dispersed phase is necessary. The formulation is a water-in-oil type emulsion (W/O), the dispersed phase is a concentrated solution of polysaccharide. The polysaccharide solution is devoted to be released during the preparation of the phytosanitary bulk in a spray tank. The formulation should have stability over time complying with the regulations for phytosanitary liquid formulations. Therefore, the stability of the emulsion was optimized using the appropriate composition (continuous phase, dispersed phase, surfactant) and an optimized manufacturing process using a rotorstatorsystem and a high pressure homogenizer. The formulations were characterized using techniques which allow assessing the average size and size distribution of the dispersed droplets such as laser light diffraction granulometry and differential scanning calorimetry (DSC). The crystallization temperature of the droplets measured during the DSC analysis is correlated to their size. The stability over time was assessed using natural ageing and accelerated ageing tests. The release of polysaccharide was then studied in order to be performed under the conditions of preparing of phytosanitary bulk by a farmer. The release is achieved by a prior destabilization of the formulation using a demulsifier surfactant. The release is evaluated by the evolution of the electrical conductivity of the bulk, as thepolysaccharides entrapped form ionized molecules in aqueous media. The study of the spray has subsequently been carried out using a pilot reproducing an agricultural spray boom. Spraying was optimized by varying the physicochemical properties of the bulk preparation (surface tension and rheology) and spraying parameters (pressure, spray nozzle).The retained adjuvant formulation is an emulsion containing 50 % (v/v) of solution of 3 % (w/w) of carboxymethylcellulose, dispersed in an oil phase consisting of rapeseed methyl ester with 10 % (w/w) of polyglycerol polyricinoleate. The release is achieved with an ethoxylated surfactant, selected among seven commercial surfactants. The rheological study of the spray formulation exhibits properties which limit the formation of drops of small diameter, confirmed by the tests performed on the spray pilot.

<p>No</p> / Dissertation

La vaccination est un moyen de prévention très efficace contre les infections. La majorité des vaccins sont administrés par voie sous-cutanée ou intra-musculaire avec des adjuvants et stimulent ainsi des réponses immunitaires adaptatives systémiques. Les muqueuses représentent une porte d’entrée majeure pour de nombreux pathogènes ayant un impact en Santé Publique. Cependant, peu de vaccins sont délivrés par les muqueuses en raison du manque d’adjuvants adaptés à ces voies. Ainsi le développement d'adjuvants muqueux permettrait la stimulation de réponses immunitaires locales et une protection efficace avant la dissémination des pathogènes. Les agonistes des Toll-Like-Receptors (TLR) sont développés comme adjuvants vaccinaux car ils stimulent l’immunité innée et adaptative. Au laboratoire, la flagelline de Salmonella enterica qui est un puissant agoniste de TLR5 est utilisée comme modèle afin de disséquer les mécanismes d’action des adjuvants muqueux. En effet, l'administration intranasale de vaccins adjuvantés par la flagelline se caractérise par une réponse T CD4+ de type Th1/Th2, une réponse en anticorps sécrétoires dans le compartiment respiratoire et une réponse systémique contre les antigènes vaccinaux. Seule l’activation de TLR5 dans le compartiment épithélial est nécessaire à l’activité adjuvante.Dans un premier temps, l'analyse transcriptionnelle du tissu pulmonaire a permis d'identifier une signature épithéliale spécifique du recrutement de cellules immunitaires, en particulier de monocytes et de neutrophiles ainsi que de l'activation fonctionnelle des cellules dendritiques. L'analyse de la dynamique cellulaire au niveau du tractus respiratoire et des ganglions drainants a ensuite été réalisée en réponse à l’administration intranasale de vaccin adjuvanté. Bien que les monocytes inflammatoires et les neutrophiles infiltrent massivement les poumons et capturent les antigènes, ils ne jouent pas de rôle majeur dans l’activation de la réponse immunitaire. Au contraire les cellules dendritiques conventionnelles CD11b+ capturent, migrent et présentent efficacement l’antigène aux lymphocytes T CD4+. A l'instar de l’effet adjuvant, l’activation de ces cellules dendritiques par la flagelline n'est pas directe mais requiert une expression de TLR5 dans les cellules structurales incluant les cellules épithéliales de la muqueuse. De plus, nos travaux suggèrent que les interleukines de la famille IL-1 ne sont pas à l'origine de la transactivation des cellules dendritiques.En conclusion, ce travail de thèse ouvre des perspectives intéressantes quant au développement d’adjuvants muqueux. / Many pathogens of public health concern (including the influenza and respiratory syncytial viruses, and bacteria such as Streptococcus pneumoniae and Pseudomonas aeruginosa) enter the body via the respiratory tract in general and the lung mucosa in particular. Mucosal vaccines induce a local adaptive immune response (i.e. secretory antibodies and specific T cells) and constitute a unique means of directly preventing these infections. Most vaccines are delivered systemically and use systemic adjuvants. Although the few commercially available mucosal vaccines are generally effective, mucosal adjuvant candidates have not demonstrated sufficient levels of potency and safety. TLR signaling is instrumental for the induction of innate immunity and the concomitant ignition of adaptive immune responses. Thus TLR agonists are largely used as vaccine adjuvants. In the lab we use flagellin from Salmonella enterica (a potent TLR5 agonist) as a model to better understand the mode of action of mucosal adjuvants. The intranasal adjuvant effect of flagellin is characterized by an antigen-specific Th1/Th2 cell response, and a strong mucosal and systemic antibody response. However this adaptive immune response mainly depends on TLR5-mediated epithelial signaling.We used molecular profiling to show that cytokine/chemokine and dendritic cell maturation pathways are surrogate signatures for flagellin activation in the lung. Neutrophils and inflammatory monocytes were massively recruited to the lungs but were not essential for the adjuvant activity. In contrast, flagellin signaling did not induce a significant recruitment of conventional dendritic cells but enhanced their maturation and migration to the lymph nodes. In particular, CD11b+ migratory dendritic cells were essential for induction of a CD4+ T-cell response. Importantly, the functional activation of dendritic cells was independent of direct signaling via TLR5, suggesting the role of inflammatory cytokines produced by the activated epithelium. However or data suggest that IL-1 and IL-36 interleukins are not responsible for transactivation of dendritic cell. In conclusion, this thesis project opens up new perspectives for the development of mucosal adjuvants.

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.

O desenvolvimento de uma vacina para o controle da dengue é uma prioridade em todo o mundo. O domínio helicase da proteína NS3 (NS3H) viral alberga epítopos reconhecidos por linfócitos T citotóxicos, os quais tem papel importante na eliminação de células infectadas. Esse trabalho propôs a obtenção de uma forma recombinante, produzida em linhagens de Escherichia coli, da NS3H do DENV2 com características similares à proteína nativa e sua utilização como um potencial antígeno vacinal. A NS3H foi obtida na forma solúvel, foi reconhecida por anticorpos de camundongos e de humanos infectados e foi capaz de interagir com o RNA viral. Camundongos imunizados com NS3H coadministrada com diferentes adjuvantes desenvolveram respostas imunológicas específicas mas não foram protegidos após desafio. Em conjunto, os resultados indicam que a proteína NS3H recombinante preserva conformação e determinantes antigênicos da proteína viral nativa e pode ser útil em estudos sobre a biologia viral e na busca de estratégias anti-virais voltadas para o controle da dengue. / The development of a dengue vaccine is a worldwide priority. The helicase domain of viral NS3 protein (NS3H) preserves epitopes recognized by cytotoxic T lymphocytes, which plays an important role in the elimination of infected cells. This study aimed the generation of a recombinant NS3H form of a type 2 dengue virus (DENV2) lineage, in Escherichia coli strains, with properties similar to the native protein and its use as a potential vaccine antigen. The NS3H was obtained in soluble form, was recognized by antibodies from mice and human subjects and was able to interact with the viral RNA. Mice immunized with NS3H combined with different adjuvants developed specific immune responses but did not confer protection to a lethal challenge. Altogether, the results indicate that the recombinant NS3H protein preserves conformational and antigenic determinants of the native protein and may be a useful tool for studies dealing with the DENV biology and the search for anti-virus approaches.

Esporos de Bacillus subtilis podem ser utilizados como veículos vacinais capazes de expressar ou adsorver proteínas heterólogas em sua superfície. Estudos recentes indicaram que esporos de B. subtilis também conferem um forte efeito adjuvante para vacinas baseadas em proteínas purificadas administradas por via parenteral. Nesse cenário, o objetivo do presente projeto foi avaliar o papel adjuvante dos esporos de B. subtilis e viabilizar sua utilização como carregadores de vacinas de DNA pelo método da biobalística (gene gun). Nesta metodologia, os esporos foram utilizados como substitutos das micropartículas de ouro usualmente empregadas, mantendo a capacidade de transfecção de células eucarióticas in vitro, e produção de anticorpos específicos em camundongos C57BL/6 imunizados com o plasmídeo vacinal pgDE7h (que codifica para uma forma atóxica da oncoproteína E7 do HPV-16). Além disso, esporos de B. subtilis foram capazes de ativar células dendríticas in vitro e, quando coadministrados a pgDE7h pela via intramuscular, conferiram um aumento de 50% na proteção anti-tumoral de animais previamente transplantados com a linhagem tumoral TC-1 (que expressa a proteína E7 do HPV-16), em uma relação dose-dependente. / Bacillus subtilis spores can be used as vaccine vehicles capable of to display or adsorb heterologous proteins in its surface. Recent studies have indicated that spores of B. subtilis confer a strong adjuvant effect for vaccines based on purified proteins administered by the parenteral route. In this scenario, the objective of this project is to evaluate the adjuvant role of B. subtilis spores e viabilize its use as DNA vaccine carriers of biolistic method (gene gun). In this methodology, the spores are used as gold microparticles substitutes usually employed, maintaining the ability to transfect eukaryotic cells in vitro and production of specific antibodies in C57BL / 6 mice immunized with the vaccine plasmid pgDE7h (encoding a non-toxic form of the E7oncoprotein of HPV-16). Additionally, spores of B. subtilis were able to activate dendritic cells in vitro when co-administered intramuscularly to pgDE7h, conferring an increase of 50% of anti-tumor protection in animals previously transplanted with the TC-1 tumor cell line (expressing the E7 protein of HPV-16) in a dose-dependent manner.

Streptococcus mutans é considerado o principal agente etiológico da cárie dental humana, uma doença crônica e de caráter infeccioso. A doença apresenta uma ampla incidência e é considerada um importante problema de saúde pública mundial. A adesão inicial de S. mutans à superfície dental é dependente da interação da proteína de superfície P1 e a aglutinina salivar adsorvida ao dente. Sendo assim, a proteína P1 e seus fragmentos são considerados importantes alvos em novas estratégias vacinais contra a cárie dental. O presente trabalho teve como objetivo desenvolver e caracterizar estratégias vacinais de mucosa contra S. mutans, em que empregamos como antígeno modelo a proteína P139-512, um fragmento N-terminal derivado da proteína P1. Para esse fim, empregamos esporos não recombinantes de Bacillus subtilis e um mutante atóxico da toxina termo-lábil (LTK63) de Escherichia coli enterotoxigência (ETEC) como adjuvantes vacinais. A proteína P139-512 foi obtida por meio de sistema de expressão baseado em linhagem recombinante de B. subtilis. As formulações vacinais foram baseadas na coadministração da proteína P139-512 com os esporos e/ou LTK63 pela via sublingual. Os resultados obtidos demostraram as propriedades imunomoduladoras dos dois adjuvantes sobre as respostas de anticorpos. No entanto, o adjuvante LTK63 foi capaz de induzir maiores títulos de anticorpos antígeno-específicos e desencadear respostas de mucosa com a indução de anticorpos IgA secretores (S-IgA) em amostras de saliva. Além disso, a formulação com esse adjuvante induziu repostas de anticorpos mais eficientes na capacidade de reconhecer a proteína P1 nativa, expressa na superfície de linhagens de S. mutans. A estratégia utilizando os dois adjuvantes não foi capaz de aumentar a magnitude das respostas de anticorpos sistêmicos e de S-IgA quando comparada à administração da P139-512 + LTK63. Em conclusão, os resultados abrem perspectivas para o desenvolvimento de estratégias vacinais de mucosa direcionadas para o controle da cárie dental baseadas na coadministração de antígenos de S. mutans com o adjuvante LTK63. / Streptococcus mutans is considered the major etiological agent of human dental caries, a chronic and infectious disease. The disease has a broad incidence and is considered an important worldwide public health problem. The initial adhesion of S. mutans to the tooth surface depends on the interaction of the surface-exposed P1 protein and the salivary agglutinin adsorbed to the tooth. Therefore, the P1 protein and fragments derived from it are considered important targets in vaccine strategies to prevent dental caries. The present work aimed to develop and characterize new mucosal vaccine strategies against S. mutans, using as antigen model the protein P139-512, a N-terminal fragment derived from the P1 protein. For this purpose, we used non-recombinant Bacillus subtilis spores and a non-toxic mutant of the enterotoxigenic Escherichia coli (ETEC) heat-labile toxin (LTK63) as vaccine adjuvants. The P139-512 protein was obtained using a recombinant B. subtilis strain. Vaccine formulations were based on the co-administration of the P139-512 protein with spores and/or LTK63 via sublingual route. The results demonstrated that both adjuvants showed immunomodulatory properties on antibody responses. However, the LTK63 adjuvant was capable to induce the highest antigen-specific antibody titers and elicited mucosal responses with the induction of secretory IgA (S-IgA) antibodies in saliva samples. In addition, the formulation using this adjuvant induced enhanced responses mediated by effector antibodies capable to recognize the native P1 protein expressed on the surface of S. mutans strains. The combination of both adjuvants did not increase the magnitude of systemic antibody and S-IgA responses when compared to the administration of P139-512 + LTK63. In conclusion, the results open perspectives for the development of vaccination strategies for the control of dental caries based on the co-administration of S. mutans antigens and LTK63.

No interesse de reduzir os efeitos adversos relacionados às vacinas veterinárias e induzir tipos específicos de resposta imune, levou-se o desenvolvimento de numerosos novos adjuvantes. Recentemente, houve o descobrimento de um peptídeo intracelular funcional obtido através do estímulo celular com INF- &#947;, denominado EL28, tendo como principais características a hidrossolubilidade e a biodegradebilidade e, porém, no momento carece de ensaios de imunoestimulação associados com vacinas com potencial de mercado \"in vivo\". Neste sentido, o presente estudo teve como objetivo verificar a atividade imunoestimulante do EL28, associado ao vírus rábico inativado (VRI). Realizou-se a produção do antígeno VRI, e prepararam-se formulações vacinais: VRI associado ao EL28, VRI associado com Al(OH)3 e VRI sem presença de adjuvante, para imunização de camundongos Balb/c e avaliação da resposta imune humoral. A produção de anticorpos IgG totais, IgG1 e IgG2a foram determinados pelo ensaio de ELISA indireto. E os anticorpos neutralizantes para o vírus rábico foram quantificados através do método RIFFT. Os resultados do ELISA indireto foram expressos como Dose Imunoenzimática 50% (DI50) e mostraram que não houve aumento na produção de anticorpos na resposta imune humoral (IgG totais) entre a vacina associada ao EL28 (DI50= 3,0 X 10-3&#181;L) e a vacina sem adjuvante (DI50=2,0 X 10-3&#181;L). As IgG2a e IgG1 estão relacionados ao perfil da resposta Th1 e Th2 respectivamente, e mostraram diferença significativa em relação ao título da IgG1 entre a vacina associada ao EL28 (DI50= 5,9x10-4&#181;L) e a vacina sem adjuvante (DI50=5,2x10-4&#181;L), enquanto para os títulos de IgG2a não houve diferença significativa entre essas vacinas. O perfil da resposta Th foi obtido através da razão entre IgG2a e IgG1, o qual demonstrou que em todas as imunizações, com e sem adjuvantes, tiveram resposta predominante do tipo Th2. O método de RIFFT mostrou um aumento na potência de anticorpos neutralizantes na vacina associada ao EL28 em relação à imunização dos animais sem nenhum adjuvante. Portanto, esses resultados demonstram aumento da especificidade na resposta imunológica da vacina associada ao EL28. Sendo assim, pelos resultados apresentados, a vacina de raiva associada ao EL28 possivelmente teria uma maior eficiência e segurança. / In the interest of reducing the adverse effects related to veterinary vaccines and inducing specific types of immune response, numerous new adjuvants have been developed. Recently, a functional intracellular peptide obtained through cellular stimulus with the INF-&#947;, named EL28, it have as main characteristics the water solubility and the biodegradability, but at the moment it lacks immunostimulation assays associated with potencial market vaccine \"in vivo\". In this sense, the present study aimed to verify the immunostimulatory activity of EL28, associated with inactivated rabies virus (IRV). It carried out the production of IRV antigen, and prepared vaccine formulations: IRV with EL28, IRV with Al(OH)3 and IRV without the presence of adjuvant, for immunization of Balb/c mice and evaluation of the humoral immune response. The production of total IgG antibodies, IgG1 and IgG2a were determined by the indirect ELISA assay. And neutralizing antibodies to rabies virus were quantified using the RIFFT method. Indirect ELISA results were expressed as 50% Immunoenzymatic Dose (ID50) and showed no increase in antibody humoral immune response (IgG total) between the EL28 associated vaccine (ID50 = 3.0 X 10-3&#181;L) and the vaccine without adjuvant (ID50 = 2.0 X 10-3&#181;L). The IgG2a and IgG1 are related to the profile of Th1 and Th2 respectively, and showed a significant difference in relation to the IgG1 titer between the EL28 associated vaccine (ID50 = 5,9x10-4&#181;L) and the non-adjuvant vaccine (ID50 = 5,2x10-4&#181;L), while for IgG2a titers there was no significant difference between these vaccines. The Th response profile was obtained by the ratio of IgG2a and IgG1, which showed that in all immunizations with and without adjuvant, were predominantly Th2 response. The RIFFT method showed an increase of the potency of neutralizing antibodies in the vaccine with EL28 in relation to the immunization of the animals without any adjuvant. Therefore, these results demonstrate increased specificity in the immune response associated with EL28 vaccine. Therefore, from the results presented, the rabies vaccine associated with EL28 would possibly have a greater efficiency and safety.

Paracoccidioidomicose (PCM) é uma doença sistêmica granulomatosa causada pelo fungo termo-dimórfico Paracoccidioides brasiliensis. A gp43 é o principal antígeno diagnóstico que contém o peptídeo de 15 aminoácidos designado como P10, que desenvolve a resposta imune do tipo Th1 dependente de IFN-<font face=\"Symbol\">g. No presente trabalho, comparamos a efetividade de diferentes adjuvantes em camundongos BALB/c infectados intratraquealmente com o isolado virulento Pb18. Alúmen, Adjuvante Completo e Incompleto de Freund, FliC flagelina e o lipídeo catiônico, foram testados em associação ou não com P10. Após 52 dias de infecção, o lipídeo catiônico mostrou os melhores resultados, com o menor número de UFCs, redução de fibrose e baixo número de células fúngicas no tecido pulmonar, além do incremento significativo de IFN-<font face=\"Symbol\">g e TNF-<font face=\"Symbol\">a e a redução dos níveis de IL-4 e IL-10. Estes resultados sugerem que a interação do peptídeo P10 com o lipídeo catiônico pode gerar uma melhor resposta imune mediada por células do tipo Th1, evitando a rápida disseminação da PCM experimental. / Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis. The gp43 is the major diagnostic antigen that contains the peptide P10. This peptide has 15 amino acids and develops a Th1- dependent IFN-<font face=\"Symbol\">g immune response. In this work, we compare the effectiveness of different adjuvants in BALB/c mice infected intratracheally with virulent isolate Pb18. Alum, Complete Adjuvant and Incomplete Freund\'s adjuvant, fliC flagellin from Salmonella enterica and cationic lipid were tested in combination or not with P10. After 52 days of infection, the cationic lipid showed the best results with the least number of colony forming units, reduction of fibrosis, low number of fungal cells in lung tissue and the significant increase of IFN-<font face=\"Symbol\">g and TNF-<font face=\"Symbol\">a with reduced levels of IL-4 and IL-10. These results suggest that, the interaction of peptide P10 with cationic lipids can lead to a better immune response mediated by Th1 type cells by avoiding the rapid spread of experimental PCM.