ABSTRACT Although there is ample evidence linking chronic inflammation with cancer, the cellular mechanisms involved in early events leading to tumor development remain unclear. Myeloid cells are an intricate part of inflammation. They consist of mature cells represented by macrophages, dendritic cells and granulocytes and a population of Immature Myeloid Cells (IMC), which in healthy individuals are cells in transition to mature cells. There is a substantial expansion of IMC in cancer and many other pathological conditions which is associated with pathologic activation of these cells. As a result, these cells acquire the ability to suppress immune responses and are termed Myeloid-derived Suppressor Cells (MDSCs). Although the role of MDSC in immune suppression in cancer and tumor progression is well established, their contribution to tumor development is still uncertain. The fact that cells with MDSC phenotype and function are observed in chronic inflammation raised the possibility that these cells can contribute to initial stages of tumor development. To address this question, we used an experimental system where the number of IMC was regulated by the expression of S100A9 protein. In this project, we used two different models of chronic inflammation in S100A9 transgenic (S100A9tg) and S100A9 knock-out (S100A9KO) mice. In the first model, we created the conditions for topical accumulation of these cells in the skin in the absence of infection or tissue damage using S100A9tg mice. Accumulation of IMC in the skin resulted in a dramatic increase in the formation of skin tumors during epidermal carcinogenesis. Conversely, lack of myeloid cell accumulation in S100A9KO mice substantially reduced the formation of skin papillomas. The effect of IMC was not associated with immune suppression but with the recruitment of CD4+ T cells mediated by CCL4 chemokine released by activated IMC. Elimination of CD4+ T cells or blockade of CCL4 abrogated the increase in tumor formation caused by myeloid cells. Thus, this study implicates the accumulation of IMC as an initial step in facilitating of tumor formation, which can mediate the recruitment of CD4+ T cells via the release of CCL4 chemokine. In the second model, we used inflammation-associated lung cancer caused by the chemical lung carcinogen urethane in combination with exposure to cigarette smoke referred to throughout as CS. Exposure of mice to CS alone resulted in a significant accumulation of cells with typical MDSC phenotype in different organs; however, these cells lacked immune suppressive activity and could not be defined as bona fide MDSC. When CS was combined with the single dose of urethane, it led to the accumulation of immune suppressive cells. The expansion of MDSC followed the onset of lung tumors development. This suggests that MDSC in this model is not the preceding factor but rather a consequence of tumor formation. Further studies are necessary to determine the relevance of targeting these cells for cancer treatment and prevention.

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No. of bitstreams: 2 Dissertação - Barbara Martins Vieira - 2014.pdf: 1451459 bytes, checksum: 595a3718c8c4e83f9334cf6513f85119 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-08-22 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The carbon tetrachloride (CCl4) is recognized as a classic hepatotoxin, being considered the best method to induce liver injury, commonly used as a model to test the hepatoprotective effect of drugs and natural substances and for investigation of hepatotoxicity, cytotoxicity and oxidative stress, in both in vivo and in vitro studies. This study aimed to standardize the lowest dose of CCl4 to cause acute liver injury by oxidative stress in Wistar rats. The in vivo experiment was carried out with 12 male Wistar rats (180-240g),which were maintained for 16 days under controlled environment and supplied with water and Purina® rodent ad libitum. The animals were separated into four groups: CG - control group; G0,5 - dose of 0.5 ml / kg body weight (bw); G0,75 - dose of 0.75 ml / kg bw and G1 - dose of 1 ml / kg bw. CCl4 was administered by intraperitoneal injection twice a week. 24h after the last CCl4 administration, all animals were anesthetized (xylazine: ketamine - 1: 1 v / v) for performing cardiac puncture, euthanasia and dissection of the liver for removal and analysis. Results obtained were subjected to normality test, and followed by the analysis of variance (ANOVA) and comparison of means (Tukey at 5% probability - post-hoc). It was observed that the mean weights of rats treated with CCl4 were higher than that of to the GC and the liver protein content (in g / 100 g) was lower in the treated groups, with no statistical difference between the test groups. Histopathological analysis showed changes in the structure, increased numbers of macrophages with intracellular lipid accumulation and increased cellular infiltration in groups treated with CCl4. By quantifying the enzymes alanine aminotransferase (ALT), aspartate and alanine aminotransferase (AST), it was found that the CCl4-treated groups showed significantly higher levels than CG, being higher in G1 compared to the other groups tested. All tested concentrations of CCl4 induced liver injury in vivo, in different degrees, and the concentration of 0.5 mL / kg bw, administered twice weekly, was the lowest dose tested that could cause changes in all the parameters evaluated. Therefore, this is the ideal dose for induction of acute liver injury, aiming to test the modulatory role of dietary and nutritional factors. / O Tetracloreto de Carbono (CCl4) é reconhecido como hepatotoxina clássica, sendo considerada a melhor substância para induzir lesão hepática, comumente utilizado como modelo para testar o efeito hepatoprotetor de drogas e substâncias naturais e para investigação de hepatotoxicidade, citotoxicidade e estresse oxidativo, tanto para estudos in vivo quanto in vitro. O presente estudo visou padronizar a menor dose de CCl4 capaz de provocar lesão hepática aguda por estresse oxidativo em ratos Wistar. O experimento in vivo foi realizado com 12 ratos Wistar adultos, machos, mantidos em condições de ambiente controladas e fornecimento de água e ração distribuídos ad libitum. O ensaio teve duração de 16 dias. Os animais foram separados em quatro grupos, sendo: GC – grupo controle; G0,5 – dose de 0,5 mL/kg de peso corporal (pc); G0,75 – dose de 0,75 mL/kg de pc e G1 – dose de 1 mL/kg de pc. O CCI4 foi administrado via injeção intraperitoneal, duas vezes por semana. Após 24h da última administração de CCI4, todos os animais foram anestesiados para realização de punção cardíaca, eutanásia e dissecação para retirada do fígado e análises. Os resultados obtidos foram submetidos ao teste de normalidade e, posteriormente, à análise de variância e comparação de médias. Observou-se que o peso do fígado dos ratos tratados com CCl4 foi maior em todos os grupos tratados em comparação ao GC e o teor protéico do fígado (em g/100g) foi menor nos grupos tratados, sem diferença estatística entre os grupos teste. As análises histopatológicas mostraram alteração na estrutura do tecido hepático, aumento do número de macrófagos com acúmulo intracelular de lipídeos e aumento do infiltrado celular nos grupos tratados com CCl4. Por meio da quantificação das enzimas alanina aminotransferase (ALT) e alanina aspartato aminotransferase (AST), verificou-se que os grupos tratados com CCl4 apresentaram concentrações significativamente mais elevadas em relação ao GC, sendo maiores no G1 em comparação aos demais grupos testados. Todas as concentrações de CCl4 testadas induziram lesão hepática, em diferentes graus, sendo a concentração de 0,5 mL/kg de pc, administrada duas vezes por semana, a menor dose testada que conseguiu provocar alterações em todos os parâmentos avaliados. Portanto, esta é a dose mais indicada para indução de lesão hepática aguda, objetivando testar o papel modulador de fatores alimentares e nutricionais.

Glyoxalase-I (Glo-I) and glyoxalase-II (Glo-II) comprise the glyoxalase system and are responsible for the detoxification of methylglyoxal (MGO). MGO is formed non-enzymatically as a by-product, mainly in glycolysis, and leads to the formation of advanced glycation endproducts (AGEs). AGEs bind to their receptor, RAGE, and activate intracellular transcription factors, resulting in the production of pro-inflammatory cytokines, oxidative stress, and inflammation. This review will focus on the implication of the Glo-I/AGE/RAGE system in liver injury and hepatocellular carcinoma (HCC). AGEs and RAGE are upregulated in liver fibrosis, and the silencing of RAGE reduced collagen deposition and the tumor growth of HCC. Nevertheless, data relating to Glo-I in fibrosis and cirrhosis are preliminary. Glo-I expression was found to be reduced in early and advanced cirrhosis with a subsequent increase of MGO-levels. On the other hand, pharmacological modulation of Glo-I resulted in the reduced activation of hepatic stellate cells and therefore reduced fibrosis in the CCl4-model of cirrhosis. Thus, current research highlighted the Glo-I/AGE/RAGE system as an interesting therapeutic target in chronic liver diseases. These findings need further elucidation in preclinical and clinical studies.

Os compostos fenólicos são substâncias amplamente distribuídas no reino vegetal, em particular nas frutas e em outros vegetais. Estes compostos, destacando-se flavonoides e os ácidos fenólicos, devido à estrutura molecular, podem apresentar a capacidade de inibir processos oxidativos. Além do mais, estão relacionados com a redução de risco de doenças crônicas não transmissíveis tais como: cardiovasculares, câncer, aterosclerose, entre outras. Considerando a elevada produção de caju em território brasileiro e a possibilidade da existência de compostos com potencial antioxidante no pedúnculo de caju , este trabalho teve por objetivo avaliar quantitativa e qualitativamente os compostos fenólicos, particularmente os ácidos fenólicos, e identificar a participação destes em processos metabólicos do organismo animal. Foram caracterizados quimicamente três clones distintos de pedúnculos de caju (CCP-76, CCP-09, BRS-189 e CCP-76 tratado) e na análise química, apresentaram um elevado teor de ácidos graxos monoinsaturados, predominando o ácido oléico, e de fenólicos totais. Os ácidos fenólicos identificados foram: gálico, protocatecuíco, p-cumárico, ferúlico, caféico e salicílico. Foram obtidos extratos aquoso (EAq) e alcoólico (EAlc) e frações de ácidos fenólicos a partir dos pedúnculos e, avaliados em sistemas modelo β-caroteno/ácidolinoléico e em Rancimat. As frações de ácidos fenólicos exibiram expressiva atividade antioxidante no primeiro sistema e os extratos, no segundo, demonstraram fatores de proteção superior ao antioxidante sintético BHT. Pôde-se também verificar a capacidade antioxidante dos extratos e frações do clone CCP-76 no sistema de varredura do radical DPPH. Em ensaio experimental com ratos, em condição normal, foi administrado EAq (80 e 240 mg/kg, v.o.) ou fração de ácidos fenólicos livres (40 e 120 mg/kg, v.o.) obtidos do pedúnculo de caju CCP-76. Neste estudo, não se observou potencialização de todos os antioxidantes enzimáticos (superóxido dismutase, catalase e glutationas peroxidase e redutase), contudo pôde-se verificar a redução dos níveis de lipoperoxidação no tecido cerebral dando indícios de aumento do estado antioxidante nos animais. Também foi avaliado o potencial antioxidante do EAq e da fração de ácidos fenólicos livres sobre o dano hepático em ratos tratados com tetracloreto (CCl4) de carbono. A administração deste teve seu efeito corroborado pela avaliação dos parâmetros bioquímicos, ou seja, aumento exacerbado das enzimas hepáticas no plasma: alanina transaminase (ALT) e aspartato transaminase (AST); decréscimo da atividades da enzimas antioxidantes no fígado e elevação da produção de peróxidos lipídicos no tecido hepático. Nos ratos que receberam EAq (480 mg/kg, v.o.) não se observou alteração comparando-os aos animais tratados apenas com CCl4 . No entanto, a administração de fração de ácidos fenólicos livres, nas duas doses (40 e 120 mg/kg, v.o.), evidenciou pronunciado efeito contra a lesão hepática, com níveis reduzidos de AL T e AST plasmáticas, aumento da atividade das enzimas antioxidantes no fígado e prevenindo a lipoperoxidação hepática mediada pelo radical CCl3• a partir do CCl4. Estudos histológicos do tecido hepático confirmaram as avaliações bioquímicas exibindo preservação tecidual, supressão de degeneração vacuolar macro e microgoticular e de sinais necróticos nos ratos tratados com a fração de ácidos fenólicos livres do pedúnculo de caju. / Phenolic compounds are widely distributed in the plant kingdom, particularly fruits and vegetables. Due to their chemical structure, these compounds, in particular flavonoids and phenolic acids, are able to inhibit oxidative processes. Furthermore, can be used to reduce the risk of non-transmissible chronic diseases such as cardiovascular diseases, cancer and atherosclerosis. Taking into consideration the large production of cashew in Brazil and the possible existence of potentially antioxidant compounds present in the cashew apples, the aim of this study was to quantitatively and qualitatively evaluate the presence of phenolic compounds in cashew apple, particularly phenolic acids, and identify their role in metabolic processes in animals. The cashew apples of three distinct clones (CCP-76, CCP-09, BRS-189 and CCP-76 (processed)) were studied. The determination of fatty acids yielded a high concentration of monounsaturated fatty acids, mainly oleic acid, and of total phenolic compound. The phenolic acids found were: gallic, proteocatechuic, p-cumaric, ferulic, caffeic and salicylic acids. Both aqueous (EAq) and ethanolic (EAlc) extracts and phenolic acid fractions were obtained from the cashew apples and were evaluated in a β-carotene/linoleate model system and Rancimat test. The phenolic acid fractions presented an expressive antioxidant activity in the β-carotene/linoleate model system and the extracts, by the Rancimat test presented a protection factor higher than that of antioxidant additive, BHT. We also observed the antioxidant capacity of the extracts and fractions of the CCP-76 clone in the DPPH radical scavenging assay. In an experimental assay with rats, the EAq (80 and 240 mg/kg) or the free phenolic acid fraction (40 and 120 mg/kg) obtained from the cashew apple of CCP-76 clone was administered via the oral route. In this study, the enhancement of enzymatic antioxidants (superoxide dismutase, catalase, glutathione peroxidase and reductase) was not observed, nevertheless, a decrease in the amount of lipoperoxidation in the brain tissue was observed, suggesting that the ingestion of cashew might increase the antioxidative state in animals. Also, the antioxidant activity of EAq and of the free phenolic acid fraction from the cashew apple of CCP-76 clone was verified on the liver damage induced by carbon tetrachloride. The liver damage caused by the administration of carbon tetrachloride was detected by biochemical parameters, namely, the increase in the serum concentrations of alanine transaminase (ALT) and aspartate transaminase (AST) as well as a decrease in the activities of antioxidant enzymes and an increase in peroxidation in the liver. Rats who received EAq (480 mg/kg, p.o.) did not present alterations in any of the parameters evaluated, compared to the animals treated with carbon tetrachloride. On the other hand, the administration of free the phenolic acid fraction in doses of 40 and 120 mg/kg, p.o., had a pronounced effect in protecting against hepatic lesion, which was evidenced by the decrease in plasma ALT and AST, enhancing the activity of antioxidant enzymes and preventing lipoperoxidation mediated by the CCl3• radical generated by carbon tetrachloride. Histological studies were able to confirm the biochemical alterations observed in that the liver tissue obtained from rats treated with phenolic acid fractions extracted from cashew apple of CCP-76 clone presented a preserved tissue structure and suppression of macro and microgoticular vacuolar degeneration as well as of signs of necrosis.

Os compostos fenólicos são substâncias amplamente distribuídas no reino vegetal, em particular nas frutas e em outros vegetais. Estes compostos, destacando-se flavonoides e os ácidos fenólicos, devido à estrutura molecular, podem apresentar a capacidade de inibir processos oxidativos. Além do mais, estão relacionados com a redução de risco de doenças crônicas não transmissíveis tais como: cardiovasculares, câncer, aterosclerose, entre outras. Considerando a elevada produção de caju em território brasileiro e a possibilidade da existência de compostos com potencial antioxidante no pedúnculo de caju , este trabalho teve por objetivo avaliar quantitativa e qualitativamente os compostos fenólicos, particularmente os ácidos fenólicos, e identificar a participação destes em processos metabólicos do organismo animal. Foram caracterizados quimicamente três clones distintos de pedúnculos de caju (CCP-76, CCP-09, BRS-189 e CCP-76 tratado) e na análise química, apresentaram um elevado teor de ácidos graxos monoinsaturados, predominando o ácido oléico, e de fenólicos totais. Os ácidos fenólicos identificados foram: gálico, protocatecuíco, p-cumárico, ferúlico, caféico e salicílico. Foram obtidos extratos aquoso (EAq) e alcoólico (EAlc) e frações de ácidos fenólicos a partir dos pedúnculos e, avaliados em sistemas modelo β-caroteno/ácidolinoléico e em Rancimat. As frações de ácidos fenólicos exibiram expressiva atividade antioxidante no primeiro sistema e os extratos, no segundo, demonstraram fatores de proteção superior ao antioxidante sintético BHT. Pôde-se também verificar a capacidade antioxidante dos extratos e frações do clone CCP-76 no sistema de varredura do radical DPPH. Em ensaio experimental com ratos, em condição normal, foi administrado EAq (80 e 240 mg/kg, v.o.) ou fração de ácidos fenólicos livres (40 e 120 mg/kg, v.o.) obtidos do pedúnculo de caju CCP-76. Neste estudo, não se observou potencialização de todos os antioxidantes enzimáticos (superóxido dismutase, catalase e glutationas peroxidase e redutase), contudo pôde-se verificar a redução dos níveis de lipoperoxidação no tecido cerebral dando indícios de aumento do estado antioxidante nos animais. Também foi avaliado o potencial antioxidante do EAq e da fração de ácidos fenólicos livres sobre o dano hepático em ratos tratados com tetracloreto (CCl4) de carbono. A administração deste teve seu efeito corroborado pela avaliação dos parâmetros bioquímicos, ou seja, aumento exacerbado das enzimas hepáticas no plasma: alanina transaminase (ALT) e aspartato transaminase (AST); decréscimo da atividades da enzimas antioxidantes no fígado e elevação da produção de peróxidos lipídicos no tecido hepático. Nos ratos que receberam EAq (480 mg/kg, v.o.) não se observou alteração comparando-os aos animais tratados apenas com CCl4 . No entanto, a administração de fração de ácidos fenólicos livres, nas duas doses (40 e 120 mg/kg, v.o.), evidenciou pronunciado efeito contra a lesão hepática, com níveis reduzidos de AL T e AST plasmáticas, aumento da atividade das enzimas antioxidantes no fígado e prevenindo a lipoperoxidação hepática mediada pelo radical CCl3• a partir do CCl4. Estudos histológicos do tecido hepático confirmaram as avaliações bioquímicas exibindo preservação tecidual, supressão de degeneração vacuolar macro e microgoticular e de sinais necróticos nos ratos tratados com a fração de ácidos fenólicos livres do pedúnculo de caju. / Phenolic compounds are widely distributed in the plant kingdom, particularly fruits and vegetables. Due to their chemical structure, these compounds, in particular flavonoids and phenolic acids, are able to inhibit oxidative processes. Furthermore, can be used to reduce the risk of non-transmissible chronic diseases such as cardiovascular diseases, cancer and atherosclerosis. Taking into consideration the large production of cashew in Brazil and the possible existence of potentially antioxidant compounds present in the cashew apples, the aim of this study was to quantitatively and qualitatively evaluate the presence of phenolic compounds in cashew apple, particularly phenolic acids, and identify their role in metabolic processes in animals. The cashew apples of three distinct clones (CCP-76, CCP-09, BRS-189 and CCP-76 (processed)) were studied. The determination of fatty acids yielded a high concentration of monounsaturated fatty acids, mainly oleic acid, and of total phenolic compound. The phenolic acids found were: gallic, proteocatechuic, p-cumaric, ferulic, caffeic and salicylic acids. Both aqueous (EAq) and ethanolic (EAlc) extracts and phenolic acid fractions were obtained from the cashew apples and were evaluated in a β-carotene/linoleate model system and Rancimat test. The phenolic acid fractions presented an expressive antioxidant activity in the β-carotene/linoleate model system and the extracts, by the Rancimat test presented a protection factor higher than that of antioxidant additive, BHT. We also observed the antioxidant capacity of the extracts and fractions of the CCP-76 clone in the DPPH radical scavenging assay. In an experimental assay with rats, the EAq (80 and 240 mg/kg) or the free phenolic acid fraction (40 and 120 mg/kg) obtained from the cashew apple of CCP-76 clone was administered via the oral route. In this study, the enhancement of enzymatic antioxidants (superoxide dismutase, catalase, glutathione peroxidase and reductase) was not observed, nevertheless, a decrease in the amount of lipoperoxidation in the brain tissue was observed, suggesting that the ingestion of cashew might increase the antioxidative state in animals. Also, the antioxidant activity of EAq and of the free phenolic acid fraction from the cashew apple of CCP-76 clone was verified on the liver damage induced by carbon tetrachloride. The liver damage caused by the administration of carbon tetrachloride was detected by biochemical parameters, namely, the increase in the serum concentrations of alanine transaminase (ALT) and aspartate transaminase (AST) as well as a decrease in the activities of antioxidant enzymes and an increase in peroxidation in the liver. Rats who received EAq (480 mg/kg, p.o.) did not present alterations in any of the parameters evaluated, compared to the animals treated with carbon tetrachloride. On the other hand, the administration of free the phenolic acid fraction in doses of 40 and 120 mg/kg, p.o., had a pronounced effect in protecting against hepatic lesion, which was evidenced by the decrease in plasma ALT and AST, enhancing the activity of antioxidant enzymes and preventing lipoperoxidation mediated by the CCl3• radical generated by carbon tetrachloride. Histological studies were able to confirm the biochemical alterations observed in that the liver tissue obtained from rats treated with phenolic acid fractions extracted from cashew apple of CCP-76 clone presented a preserved tissue structure and suppression of macro and microgoticular vacuolar degeneration as well as of signs of necrosis.

No description available.

Nef is an accessory protein encoded by human and simian immunodeficiency viruses (HIV and SIV), and is critical for viral pathogenicity in vivo.The structure of Nef has been resolved and the major cellular activities of Nef are generally described as down-regulation of cell surface molecules, enhancement of virus infectivity and regulation of cell signaling and activation. Macrophages represent a key target of HIV-1 infection and may contribute significantly to viral pathogenesis by facilitating viral propagation, maintaining a viral reservoir and regulating viral replication. During HIV-1 infection, various cytokines and chemokines are induced for viral advantages more than for host defense. We have previously demonstrated that HIV-1 Nef regulates the release of chemokines, MIP-1α and MIP-1ß, from infected macrophages and have proposed that this may enhance conditions for viral replication by promoting recruitment of substrate lymphocytes to sites of infection (1). However, the molecular basis for this Nef activity remains to be defined. The main goals of this thesis are to identify the functional motif in Nef that is responsible for chemokine induction in macrophages and to elucidate the relevance of this motif to other Nef functions. Using a mutagenesis approach, we have eventually identified a novel motif (KEK) that regulates chemokine production in infected macrophages after we excluded several previously described Nef motifs. This motif is conserved in both HIV-1 and SIV Nef proteins. Mutations in this domain abrogated MIP-1ß induction as well as the Nef-dependent release of other secretory factors by macrophages. However, disruption of this motif did not affect other Nef-ascribed activities such as CD4 and MHC-I down-regulation. In addition, we have determined the involvement of viral Env proteins in Nef-induced chemokine production. Distinct signaling pathways that regulate chemokine release in macrophage will also be described. Finally, several possible roles of the KEK motif are proposed and some preliminary results of co-immunoprecipitation experiments will be presented which aim to characterize cellular proteins involved in chemokine regulation by Nef. Collectively, our studies reveal a specific determinant within Nef that is critical for chemokine release by Nef. Identification of this motif paves the way for future studies to explore the molecular machanisms of Nef-regulated cell signaling pathways. Such knowledge may point to new therapeutic strategies that interrupt Nef function and limit the course of HIV-1 infection.

La réponse immunitaire est régulée spatialement et temporellement. Les cellules immunitaires font partie d’une plus grande communauté de populations cellulaires interconnectées qui coordonnent leurs actions par la signalisation intercellulaire. Suivant une blessure hépatique, la distribution et la composition du compartiment immunitaire évoluent rapidement au fil du temps. Par conséquent, l’information sur la position des cellules immunitaires dans le tissu hépatique est essentielle à la bonne compréhension de leurs fonctions dans la santé et la maladie. Cependant, l’organisation spatiale des cellules immunitaires en réponse à une atteinte hépatique aiguë, ainsi que les conséquences fonctionnelles de leur distribution topographique spécifique, restent mal comprises. Les macrophages hépatiques sont des cellules effectrices clés pendant l’homéostasie et en réponse à des blessures, et sont impliqués dans la pathogenèse de plusieurs maladies du foie. L’hétérogénéité et plasticité des macrophages dans le foie a été exposée avec l’émergence du séquençage de l’ARN, la cytométrie en flux et la cytométrie de masse. Ces techniques ont sensiblement contribué à la compréhension de l’origine, et fonctions des macrophages dans le foie. Cependant, ces technologies impliquent la destruction du tissu pour la préparation de suspension cellulaires ce qui entraîne une perte d’information spatiale et de contexte tissulaire. Par conséquent, la caractérisation spatiale et temporelle des macrophages dans le tissu hépatique pendant l’homéostasie tissulaire, et en réponse à une blessure, fournit une nouvelle information sur la façon dont les macrophages se rapportent aux cellules voisines et leur comportement pendant les réponses immunitaires. Dans la première partie de cette étude, nous avons conçu une stratégie pour le phénotypage spatial des cellules immunitaires hépatiques dans des échantillons de tissus. Cette stratégie combine techniques d'imagerie et l’alignement numérique des images pour surmonter les limitations actuelles du nombre de marqueurs pouvant être visualisés simultanément. En outre, nous avons généré des protocoles pour la quantification automatisée des cellules d’intérêt dans des sections de tissus pour réduire la subjectivité associée à la quantification par inspection visuelle, et pour augmenter la surface et la vitesse de l’analyse. Par conséquent, un plus grand nombre de populations de cellules immunitaires ont été visualisées, quantifiées et cartographiées, et leurs relations spatiales ont été déterminées. Dans la deuxième partie de l’étude, nous avons déterminé la cinétique et la dynamique spatiale des cellules de Kupffer (KCs) et des macrophages dérivés de monocytes (MoMFs) en réponse à une atteinte hépatique aiguë au CCl4, afin de mieux comprendre leurs rôles fonctionnels, et la répartition du travail entre eux. Nous avons constaté que les KC et les MoMFs présentent des différences au niveau de la distribution tissulaire, la morphologie, et la cinétique. En plus, seulement les KCs ont proliféré pour repeupler la population de macrophages résidents pendant la réparation tissulaire. Finalement, nous avons montré que le degré de colocalization de KCs et des MoMFs avec les cellules stellaires est différent. En plus, cette colocalisation varie avec la progression de la réponse immunitaire. Dans l’ensemble, nous avons montré que les KCs et les MoMFs ont des profils spatiaux et temporels différents en réponse à une atteinte hépatique aiguë. Dans l’ensemble, les observations faites dans cette étude suggèrent que le comportement spatial et temporel d’une sous-population donnée de cellules immunitaires est distinct et sous-tend sa capacité à remplir ses fonctions spécifiques pendant la réponse immunitaire. / The immune response is spatially and temporally regulated. Immune cells are part of a larger community of interconnected immune and non-immune cell populations that coordinate their actions mostly through cell-cell intercellular signaling. In the liver, the distribution pattern, and the composition of the immune compartment evolve during an immune response to injury influencing disease pathology, progression, and response to treatment. Hence, information on the location and interacting partners of immune cells in the hepatic tissue is critical for the proper understanding of their functions in health and disease. However, the spatial organization of hepatic resident and infiltrating immune cells in response to acute injury, and the functional consequences of their specific topographical distribution, remain poorly defined. Hepatic macrophages are key effector cells during homeostasis and in response to injury and are involved in the pathogenesis of several liver diseases. The heterogeneity and plasticity of the macrophage compartment in the liver have only recently started to be appreciated with the emergence of RNA sequencing, flow cytometry, and mass cytometry. Detailed transcriptomic and phenotypic profiling have deeply expanded our understanding of macrophage biology. However, these technologies involve tissue disruption with loss of spatial information and tissue context. Therefore, the spatial and temporal profiling of liver macrophages in tissue samples during the steady state, and in response to injury, provide novel information on how the macrophages relate to neighboring cells and their behavior during immune responses. In the first part of this study, we designed a strategy for the spatial phenotyping of hepatic immune cells in tissue samples. This strategy combined serial and sequential labeling, and digital tissue alignment to overcome current limitations in the number of markers that can be simultaneously visualized. In addition, we generated protocols for automated quantification of cells of interest in whole tissue sections which removed the subjectivity associated with quantification by visual inspection and greatly increased the area and the speed of the analysis. As a result, a larger number of immune cell populations were visualized, quantified, and mapped, and their spatial relations were determined in an unbiased manner. In the second part of this study, we monitored the kinetics, and spatial dynamics of resident Kupffer cells (KCs) and infiltrating monocyte-derived macrophages (MoMFs) in response to acute liver injury with CCl4, to gain insight into their functional roles, and the distribution of labor between them. KCs and MoMFs exhibited different tissue distribution patterns and cell morphology, different kinetics, and occupied neighboring but unique microanatomical tissue locations. KCs and MoMFs displayed a different capacity to replenish the macrophage pool upon acute injury, and were differentially related to hepatic stellate cells. Different kinetics and spatial profiles revealed that KCs and MoMFs have distinct spatial signatures and suggest that they perform distinct functions during the wound-healing response to acute liver injury. In summary, we optimized techniques and put together a strategy for the spatial profiling of hepatic immune cells. Then, we used this methodology to profile resident and infiltrating macrophage subpopulations to gain insight into their biology and distinct contribution to healing in response to acute liver injury. Overall, the observations made in this study suggest that the spatial and temporal behavior of a given subpopulation of immune cells underlie its ability to perform its specific functions during the immune response.