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Ação imunomoduladora da própolis na apresentação antigênicaConte, Fernanda Lopes January 2017 (has links)
Orientador: José Maurício Sforcin / Resumo: A própolis é um produto resinoso, elaborado pelas abelhas a partir de diferentes partes das plantas, e se destaca por suas inúmeras propriedades biológicas, e pela possibilidade de aplicação na indústria farmacêutica. Recentemente, nosso grupo tem estudado sua ação sobre monócitos – células do sistema fagocítico mononuclear que exercem importante função na resposta imune. Neste projeto, visamos investigar a possível ação moduladora da própolis na apresentação de antígenos por monócitos humanos, utilizando um antígeno infeccioso (subunidade B da enterotoxina termolábil de Escherichia coli - EtxB), um antígeno tumoral (MAGE-1), o ácido retinóico (AR) e lipopolissacarídeo (LPS), simultaneamente ou não a própolis, avaliando o possível efeito citotóxico dos tratamentos, a expressão de receptores celulares (TLR-2, TLR-4, HLA-DR, CD40, CD80) e a produção de citocinas (TNF-α, IL-6, IL-10 e IL-12). A modulação da autofagia foi avaliada através da expressão de genes que codificam as proteínas Beclin-1 e LC3-II, utilizando a rapamicina simultaneamente ou não a própolis. A própolis não alterou a viabilidade dos monócitos humanos e exerceu efeito citoprotetor nas células incubadas com os estímulos. A própolis manteve a expressão basal dos receptores de superfície celular; porém, em associação com MAGE-1 e LPS, diminuiu a expressão de CD40 estimulada pelos antígenos isoladamente. Em associação com AR, a própolis manteve a ação do antígeno sobre os receptores. Já a associação da própolis co... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre
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Ação do Glucantime sobre macrófagos de camundongos / Glucantime action on mice macrophageLarissa Moreira Siqueira 16 April 2014 (has links)
Os antimoniais pentavalentes, tais como o Glucantime, são geralmente usados como fármacos de primeira escolha para o tratamento das leishmanioses, no entanto seu mecanismo de ação não é completamente esclarecido. Atua contra formas amastigotas intracelulares de Leishmania sp, comprometendo o potencial redox levando danos ao DNA do parasito. Alguns trabalhos sugerem que o Glucantime aumenta a capacidade fagocítica e a produção de TNF-alfa por fagócitos. O objetivo deste estudo foi avaliar a capacidade do Glucantime modular a atividade do macrófago, a principal célula hospedeira da Leishmania. Inicialmente, a toxicidade do Glucantime foi testada sobre macrófagos peritoneais de camundongos BALB/c, tratando as monocamadas in vitro por 48 horas. A viabilidade celular foi avaliada pelo método do MTT. A capacidade do Glucantime (0,1, 1 e 10 mg/ml) modular os macrófagos foi avaliada tratando as monocamadas de macrófagos peritoneais por 24 horas antes da infecção com Leishmania braziliensis. Após 48 horas de incubação com meio de cultura foi avaliado o índice de infecção por contagem. Antes e após a infecção foram analisados a produção de óxido nítrico (NO) pelo método de Griess, espécies reativas de oxigênio (EROS) por fluorimetria usando a sonda H2DCFDA e a produção de citocinas por ELISA. Para avaliar se o Glucantime seria capaz de modular macrófagos in vivo, camundongos suíços foram tratados por 5 dias consecutivos com 8 mg de Glucantime pela via intraperitoneal. Macrófagos peritôneais foram avaliados quanto a sua capacidade de controlar a infecção in vitro com L. braziliensis. Os resultados mostraram que nas concentrações até 10 mg/ml, o Glucantime não alterou a viabilidade dos macrófagos in vitro. O pré-tratamento dos macrófagos com Glucantime nas concentrações de 0.1mg/mL, 1mg/mL e 10mg/mL, foi capaz de reduzir o índice de infecção em 49%, 74% e 85%, respectivamente. Em macrófagos não infectados a produção de NO foi aumentada na concentração de 10mg/ml de Glucantime. O tratamento com 1 e 10 mg/ml de Glucantime foi capaz de aumentar significativamente a produção de EROs (p<0,05 e p<0.01, respectivamente) e a produção IL-12 (p<0,05), mas a IL-10 não foi alterada. Não houve alterações significativas desses parâmetros em relação ao controle após a infecção com L. braziliensis. Os macrófagos oriundos dos animais tratados com Glucantime foram capazes de reduzir o índice de infecção por L. braziliensis (p<0,05). Esses resultados sugerem que o Glucantime é capaz de ativar os macrófagos e esse efeito pode contribuir para o mecanismo de ação desse fármaco. / The pentavalent antimonial drugs, such as Glucantime, are generally used as first choice for the treatment of leishmaniasis, however its mechanism is not fully understood. It has activity against intracellular amastigotes of Leishmania sp, compromising the redox potential and causing damage to the DNA of the parasite. Some studies suggesting that Glucantime enhances phagocytosis and TNF-α production by phagocytes. The aim of this study is to evaluate the modulation of Glucantime on macrophages, the major host cell of Leishmania. Initially, Glucantimes toxicity was tested on peritoneal macrophages from BALB/c mice, by treating the monolayers in vitro for 48 hours. Cell viability was evaluated by MTT method. The capacity of Glucantime (0,1, 1 and 10 mg/ml) of modulate macrophages was evaluated by treating the monolayers of peritoneal macrophages for 24 hours before the infection with Leishmania braziliensis. After 48 hours of incubation with culture medium the infection index was evaluated by counting. Before and after the infection were analyzed the production of nitric oxide (NO) by Griess method, reactive oxygen species (ROS) by fluorimetry using the H2DCFDA dye and cytokines by ELISA. To evaluate if Glucantime could modulate macrophages in vivo, Swiss Webster mice were treated for 5 consecutive days with 8 mg Glucantime by intraperitoneal route. Peritoneal macrophages were evaluated about its capacity of control the in vitro infection with L. braziliensis. Results showed that until the concentration of 10 mg/ml, Glucantime did not alter the macrophages viability in vitro. The pre-treatment of macrophages with Glucantime at 0.1mg/mL, 1mg/mL and 10mg/mL was able to reduce the infection index in 49%, 74% and 85%, respectively. On non-infected macrophages the NO production was increased at 10mg/ml of Glucantime. The treatment with 1 and 10 mg/ml de Glucantime was able to significantly increase the ROS production (p<0,05 and p<0.01, respectively) and IL-12 production (p<0,05), however the IL-10 production was not altered. There were no significant changes of these parameters comparing to control after the L. braziliensis infection. The macrophages from the treated mice were capable of reduce the infection index by L. braziliensis (p<0,05). These results suggest that Glucantime is capable of activate macrophages and this effect could contribute to the mechanism of action of this drug.
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Caractérisation de l'impact de la croissance en biofilm sur l'activité probiotique de souches du genre Lactobacillus / Characterization of the impact of biofilm growth on the activity of probiotic strains of the genus LactobaccillusAoudia, Nabil 13 February 2014 (has links)
Une approche in vitro a consisté à étudier la formation de biofilm de souches d’origine du genre Lactobacillus d’intérêt probiotique. Nous nous sommes également attachés à évaluer l’impact de conditions de stress mimant l’environnement intestinal sur la formation du biofilm pour l’ensemble de ces souches. Les effets antagonistes des surnageants de cultures en biofilm ou en planctonique contre des agents pathogènes alimentaires ont été appréhendés. Non seulement toutes les souches testées forment des biofilms mais ce mode de croissance génère un effet antagoniste accentué pour certaines d’entre elles. Parmi les critères de sélection des bactéries à intérêt probiotique, les effets immunomodulateurs des probiotiques sont souvent recherchés. L. casei ATCC334 connue pour ses effets anti-inflammatoires a été retenue pour notre étude. A l’aide du modèle de lignée cellulaire THP-1 et en présence de LPS, le surnageant de culture de L. casei ATCC334 cultivée en biofilm s’est avéré présenter un effet anti-inflammatoire bien supérieur à celui des cultures planctoniques. Une approche utilisant des techniques biochimique et immunologique a permis d’identifier un des principes actifs responsable de l’effet anti-inflammatoire de cette souche. L’utilisation du modèle poisson zèbre a permis de montrer la colonisation de l’intestin des larves et de confirmer le rôle anti-inflammatoire de L. casei ATCC334 avec une diminution de la production des interleukines pro-inflammatoires et une augmentation de la production d'IL-10. Le recrutement des macrophages fluorescents mesuré en cytométrie de flux est également atténué chez la larve nourrie auparavant par le probotique en présence d’un agent inflammatoire. Le résultat majeur de cette étude est l’identification de la protéine GroEL qui contribue de façon significative à l’effet anti-inflammatoire de la souche L. casei ATCC334 lorsque qu’elle est cultivée en biofilm. / An in vitro approach was used to study biofilm formation by bacterial strains with probiotic properties and belonging to the Lactobacillus genus. We also evaluated the impact of stress conditions mimicking the intestinal environment on biofilm formation for all of these strains. The antagonistic effects of supernatants from cultures in biofilm or planktonic conditions against food-borne pathogens were apprehended. This growth mode generates an antagonistic effect accentuated for some of them. Among the selection criteria of interest probiotic bacteria, the immunomodulatory effects of probiotics are often sought. L. casei ATCC334 known for its anti-inflammatory effects was selected for our study. Using the model cell line THP-1 and in the presence of LPS, the culture supernatant of L. casei ATCC334 grown in biofilm was found to have an anti-inflammatory effect much greater than planktonic cultures. An approach using immunological and biochemical techniques has allowed the identification of the active substances responsible for the anti-inflammatory effect of this strain. Using the zebrafish model, we showed the colonization of the gut of the larvae and confirmed the anti-inflammatory role of L. casei ATCC334 with a decreased production of pro-inflammatory interleukins, and increased IL-10 production. Recruitment of fluorescent macrophages measured by flow cytometry was also mitigated in larvae fed previously by probotic in the presence of an inflammatory agent. The major result of this study is the identification of the GroEL protein that contributes significantly to anti-inflammatory effect of the strain L. casei ATCC334 when it is grown in biofilm.
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Associação entre a infecção pelo trichuris trichiura, produção de citocinas e doenças alérgicas das vias respiratórias (asma) em crianças da Região Metropolitana do Recife, PernambucoGonçales, Juliana Prado 27 February 2015 (has links)
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Previous issue date: 2015-02-27 / CAPEs / A prevalência de doenças alérgicas, como rinite e asma, é menor em países subdesenvolvidos, onde há uma maior exposição a agentes infecciosos, como os helmintos. A relação entre infecções com T. trichiura e a prevalência das doenças alérgicas e reatividade cutânea ainda não está estabelecida. Os estudos divergem quanto à alteração (potencializar ou reduzir) do quadro clínico e/ou testes cutâneos, bem como, a carga parasitária da população estudada. O estudo teve como objetivo verificar se existe diferença entre a ocorrência de asma alérgica, prick test, níveis séricos das citocinas IL-4, IL-6, IL-10, TNF-α e anticorpos IgE anti-ascaris em crianças com infecção ativa por T. trichiura. Para isto, crianças com ou sem asma foram definidas pelo questionário ISAAC, foram realizados o prick-test, parasitológico (Hoffman/Kato Katz) e coleta de sangue, que foi submetido a cultura (estimulada com PHA) e o sobrenadante coletado para a dosagens das citocinas (CBA). A prevalência de crianças com parasitológico positivo foi de 16,9 % (61/361 crianças), entre essas 27,9 % (17/61) foram positivas para infecção por Trichuris trichiura (12/17) ou co-infectadas por Trichuris trichiura/ Ascaris lumbricoides (5/17). O grupo de pacientes infectados, com ou sem asma, produziram níveis significantemente elevados para todas as citocinas em relação ao grupo controle. Além disso, o grupo dos pacientes infectados sem asma apresentou um tendência maior de produção de IL-6, TNF-alfa e IL-10 que os com asma; os pacientes infectados e asmáticos apresentaram uma menor reatividade no Prick Test quando comprado aos asmáticos não infectados. Então, a infecção por T. trichiura parece modular positivamente os níveis das citocinas TNF-α, IL-10 e IL-6, mas em pacientes asmáticos estes níveis tendem a ser controlados. As reações de hipersensibilidade cutânea imediata parece ser menos frequente em asmáticos quando infectados. Os dados levantam a possibilidade de uma modulação mútua entre asma e tricuríase, favorecendo um estado de maior cronicidade de ambas entidades de doença. / The prevalence of allergic diseases such as rhinitis and asthma is smaller in developing countries, where there is a greater exposure to infectious agents, such as helminths. The relationship between infection with T. trichiura and the prevalence of allergic diseases and skin reactivity is not yet established. Studies differ as to the nature of the change (increase or reduce) in the clinical condition and/or skin tests, as well as the parasite load of the studied population. The study aimed to determine whether there are differences between the occurrence of allergic asthma, prick test, serum levels of IL-4, IL-6, IL-10, TNF-α and anti-Ascaris IgE antibodies in children with active infection by T. trichiura. For this purpose, children with and without asthma were defined by the ISAAC questionnaire, prick-test and parasitological (Hoffman/Kato Katz) were performed and blood samples were collected, which were then subjected to culture (stimulated with PHA) and the collected supernatant for cytokines measurements (CBA). The prevalence of children with positive parasitological was 16.9% (61/361 children), and among these 27.9% (17/61) were positive for Trichuris trichiura infection (12/17) or co-infected with Trichuris trichiura/Ascaris lumbricoides (5/17). The group of infected patients, with or without asthma, produced significantly high levels for all cytokines in the control group. Furthermore, the group of patients infected without asthma showed a greater tendency of IL-6,TNF-α and IL-10 production than those with asthma; infected and asthmatic patients had a lower reactivity in Prick Test when compared to those with asthma who were uninfected. Thus, the infection with T. trichiura positively modulates the levels of TNF-α, IL-10 and IL-6 cytokines, but these levels in asthmatic patients tend to be controlled. The immediate hypersensitivity skin reactions appears to be less common in asthmatics when infected. The data raise the possibility of a mutual modulation between asthma and trichuriasis, favoring a state of chronic course on both disease entities.
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Immunodulation of inflammation in a murine pnemococcal sepsis modelMusie, Mbulaheni Edgar 01 October 2013 (has links)
Department of Microbiology / PhD (Microbiology)
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STAT5 interferes with PD-1 transcriptional activation and affects CD8+ T cell sensitivity to PD-1-dependent immunoregulation / STAT5はPD-1の転写活性化を阻害し、PD-1を介した免疫制御に対するCD8+T細胞の反応に影響を及ぼすWang, Guanning 24 January 2022 (has links)
京都大学 / 新制・課程博士 / 博士(医科学) / 甲第23609号 / 医科博第132号 / 新制||医科||9(附属図書館) / 京都大学大学院医学研究科医科学専攻 / (主査)教授 濵﨑 洋子, 教授 森信 暁雄, 教授 上野 英樹 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM
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Determination of Immunomodulatory Bioactivity Biomarkers and Mechanistic Insights in Umbilical Cord Mesenchymal Stromal CellsSiriwardena, Dylan 28 November 2018 (has links)
Detrimental immune and inflammatory responses contribute to the pathogenesis of various conditions, including Crohn’s disease, Lupus, and sepsis.1,2,3 Unfortunately, novel treatments for detrimental immune and inflammatory responses have been met with little success. Mesenchymal stromal cells (MSCs) represent a promising cellular therapy to treat immune and inflammatory disorders due to their ability to suppress the immune system. However, despite their promise, clinical trials that have employed MSC cellular therapies have produced varying and sometimes conflicting results. These discrepancies have been partially attributed to the cellular heterogeneity within MSC populations. To address these discrepancies, I performed transcriptomic and proteomic analysis of MSCs with varying immunomodulatory capacity to identify robust immunomodulatory biomarkers and gain better mechanistic insights into MSC immunomodulatory function. In this study, MSCs with differing immunomodulatory function were identified and the effect of in vitro passaging and proinflammatory induction on immunomodulatory ability was characterized. To characterize MSC immunomodulatory control mechanisms, RNA sequencing and proteomic analyses were performed on MSCs with different immunomodulatory capabilities. These analyses enabled the identification of potential immunomodulatory biomarkers and regulatory mechanisms. Finally, to test the therapeutic efficacy of immunomodulatory MSC subpopulations, I developed a humanize mouse model for sepsis. Overall, this work contributes to our understanding of MSC immunomodulation and to the development of a robust MSC cellular therapeutics.
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The immunomodulatory properties of messenchymal stem cells and their use for immunotherapy.Hoogduijn, Martin J., Popp, F., Verbeek, R., Masoodi, Mojgan, Nicolaou, Anna, Baan, C., Dehlke, M-H. January 2010 (has links)
No / There is growing interest in the use of mesenchymal stem cells (MSC) for immune therapy. Clinical trials that use MSC for treatment of therapy resistant graft versus host disease, Crohn's disease and organ transplantation have initiated. Nevertheless, the immunomodulatory effects of MSC are only partly understood. Clinical trials that are supported by basic research will lead to better understanding of the potential of MSC for immunomodulatory applications and to optimization of such therapies. In this manuscript we review some recent literature on the mechanisms of immunomodulation by MSC in vitro and animal models, present new data on the secretion of pro-inflammatory and anti-inflammatory cytokines, chemokines and prostaglandins by MSC under resting and inflammatory conditions and discuss the hopes and expectations of MSC-based immune therapy.
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Immunosuppression associée à l’enzyme interleukine-4 induced gene 1 (IL4I1) : régulation de l’expression dans les cellules humaines et rôle dans l’échappement tumoral à la réponse immune dans un modèle murin / Immunosuppression induced by Interleukin-4 Induced gene 1 (IL4I1) : Regulation of expression in human cells and role in tumor escape from the immune response in a murine modelLasoudris, Fanette 25 March 2011 (has links)
La protéine IL4I1 est une enzyme sécrétée dont l’activité L-amino acide oxydase vis-à-visde la phénylalanine inhibe la prolifération des lymphocytes T in vitro (Boulland et al, Blood 2007).Comme d’autres enzymes immunosuppressives, elle est exprimée dans les tumeurs au niveau descellules myéloïdes et/ou des cellules tumorales (Carbonnelle-Puscian et al, Leukemia 2009). Le but decette thèse a été de caractériser les conditions d’expression d’IL4I1 et de comprendre son rôle dans lecancer.Nous avons montré que les macrophages et les cellules dendritiques représentent la principalesource d’IL4I1 in vitro et dans des lésions inflammatoires chroniques. L’expression d’IL4I1 dans lesphagocytes mononucléés est induite par les interférons ou les ligands de TLR, activant respectivementSTAT1 et NF-kB, tandis que les lymphocytes B expriment des niveaux nettement plus faibles d’IL4I1sous le contrôle de la voie IL-4/STAT6 et de la voie CD40/NFkB. L’expression d’IL4I1 par des cellulesmonocytaires inhibe la production de cytokines Th1 et pourrait donc contribuer à la régulation del’inflammation Th1 in vivo.Dans un modèle murin de cancer, l’expression d’IL4I1 facilite le développement tumoral endiminuant la réponse T cytotoxique spécifique de la tumeur. Ceci est observé à des niveaux d’activitéIL4I1 proches de ceux mesurés dans des tumeurs humaines, suggérant qu’IL4I1 puisse contribuer àl’échappement des tumeurs au système immunitaire chez l’homme. Nous avons développé plusieursmutants d’IL4I1, afin d’évaluer l’impact de l’activité enzymatique versus celui de l’éventuelle liaison del’enzyme à un récepteur, dans l’effet protumoral observé. Un de ces mutants est actuellementdisponible pour une étude chez la souris.Nos résultats installent définitivement IL4I1 dans le panorama des enzymesimmunosuppressives associées au cancer et ouvrent la voie au développement d’inhibiteursspécifiques comme outils thérapeutiques / The IL4I1 protein is a secreted L-amino acid oxidase, which inhibits T cell proliferationthrough phenylalanine degradation in vitro (Boulland et al, Blood 2007). Similar to previously describedimmunosuppressive enzymes, IL4I1 is expressed in cancer by myeloid cells and/or tumor cells(Carbonnelle-Puscian et al, Leukemia 2009). The aim of this work was to characterize the cells andstimuli associated with IL4I1 expression and to decipher its role in cancer.We showed that macrophages and dendritic cells are the main source of IL4I1 in vitro and inchronic inflammatory lesions. IL4I1 expression in mononuclear phagocytes is induced by interferons orTLR ligands, which act through STAT1 and NFkB respectively. Conversely, B cells express dramaticallylower levels of IL4I1 under the control of IL-4/STAT6 and CD40/NFkB. IL4I1 expression by monocyticcells inhibits the production of Th1 cytokines and may thus contribute to Th1 inflammation control invivo.In a murine model of cancer, IL4I1 expression facilitates tumor development by depressing thetumor specific cytotoxic T cell response. This is observed for IL4I1 activity levels in the range of thosemeasured in human tumors, suggesting that IL4I1 may contribute to tumor immune escape in humans.We developed several IL4I1 mutants to discriminate the role of the enzymatic activity versus the bindingto a putative cell surface receptor in the protumor effect observed. One of these mutants is currentlyavailable for in vivo testing.Our results definitively establish IL4I1 in the family of immunosuppressive enzymes associatedwith cancer and pave the way for the development of specific inhibitors as therapeutic tools
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Le dévelopement et la modulation des réponses immunes par la bactérie intracellulaire Brucella / The induction and modulation of immune responses by the intracellular pathogen Brucella spp.Martirosyan, Anna 17 September 2012 (has links)
Les différents agents pathogènes ont développé de multiples stratégies pour contourner ou modifier les mécanismes de défense de l'hôte. La bactérie intracellulaire Brucella n'est pas une exception à la règle, car elle a développé des mécanismes qui lui permettent d'échapper à la surveillance immunitaire, persister pendant de longues périodes dans l'hôte et établir une infection chronique. En effet, Brucella responsable de la brucellose ou fièvre de Malte. La brucellose est une zoonose en réémergence ; dans cette maladie l'homme infecté représente une impasse épidémiologique. Il est indispensable de mieux connaître l'immunité développée contre Brucella qui est un excellent modèle d'étude d'autres maladies chroniques bactériennes. Le projet de thèse a été centré sur le développement et la modulation des réponses immunes par Brucella et les molécules bactériennes. Dans la première partie de cette thèse, nous avons analysé les réponses immunitaires innées et adaptatives lors de l'infection avec Brucella. D'une part, nous avons étudié l'influence des neutrophiles dans la réponse immunitaire lors de l'infection, en étudiant le cours de brucellose dans les modèles de souris neutropéniques. D'autre part, nous avons identifié et caractérisé in vivo une population des cellules CD4+ cytotoxiques au cours des infections avec Brucella. Dans la deuxième partie, nous nous sommes intéressés dans les modèles murins et humains aux propriétés immunomodulatrices des différentes molécules bactériennes telles que les glucans cycliques et des lipopolysaccharides (LPS). / Various successful pathogens have evolved multiple mechanisms to overcome or alter many normally very effective host defense mechanisms, including both innate and acquired immunity. The intracellular pathogen Brucella is not an exception to the rule as it displays mechanisms that allow it to evade immune surveillance and that are required to establish persistent infections in mammals. In this work, we studied the induction and modulation of immune responses by the intracellular bacteria Brucella and bacterial components. In the first part of this thesis, we have performed a systemic analysis of the innate and adaptive immune responses upon Brucella infection. On the one hand, we investigated the influence of polymorphonuclear neutrophils (PMNs) in the immune response during Brucella infection by exploring the course of brucellosis in antibody neutropenic mouse models. On the other hand, we identified and characterized in vivo a cytotoxic CD4+ T cell population upon Brucella abortus and Salmonella thyphimurium infections. In the second part, we focused on the immunomodulatory properties of various bacterial components such as cyclic glucans and lipopolysaccharides (LPS) both in mouse and human models.
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