The Myxococcus xanthus Dif and Frz chemosensory pathways play important roles in the regulation of gliding motility. The Dif system regulates the production of exopolysaccheride (EPS), which is essential for social motility and fruiting body formation. The Frz pathway controls reversal frequency, which is fundamental for directed movement by this surface-gliding bacterium. In addition, both pathways are involved in the chemotactic response towards several phosphatidylethanolamine (PE) species such that the Dif pathway is required for excitation while the Frz pathway is essential for adaptation. In this study we addressed three crucial questions regarding the signal processing of these two chemosensory pathways by focusing on DifA and FrzCD, the MCP homologs from their respective pathways. First, the receptor protein in the Dif pathway, DifA, lacks a perisplasmic domain, the typical signal-sensing structure. To examine whether DifA shares similar transmembrane signaling mechanism with typical transmembrane sensor proteins (MCPs and sensor kinases), we constructed a chimeric protein that is composed of the N-terminus of NarX (nitrate sensor kinase) and the C-terminus of DifA. This NarX-DifA chimera restores the DifA functionality (EPS production, agglutination, S-motility and development) to a "difA mutant in a nitrate-dependent manner, suggesting DifA shares a similar transmembrane signaling mechanism with typical MCPs and sensor kinases despite its unorthodox structure. Second, the M. xanthus chemotaxis is still controversial. It has been argued that the taxis-like response in this slowly gliding bacterium could result from physiological effects of certain chemicals. To study motility regulation by the Frz pathway, we constructed two chimeras between the N-terminus of NarX and C-terminus of FrzCD, which is the receptor protein of the Frz pathway. The two chimeras, NazDF and NazDR, are identical except that NazDR contains a G51R mutation in the otherwise wild-type NarX sensory module. This G51R mutation was shown to reverse the signaling output of a NarX-Tar chimera to nitrate. We discovered that nitrate specifically decreased the reversal frequency of NazDF-expressing cells and increased that of NazDR-expressing cells. These results show that directional motility in M. xanthus can be regulated independently of cellular metabolism and physiology. Surprisingly, the NazDR strain failed to adapt to nitrate in temporal assays, as did the wild type to known repellents. Therefore, the lack of temporal adaptation to negative stimuli is an intrinsic property in M. xanthus motility regulation. Third, the Dif and Frz pathways are both involved in the chemotactic response towards certain PE molecules such that the Dif pathway is required for excitation and while the Frz system is essential for adaptation. In addition, 12:0 PE, known to be sensed by DifA, results in increased FrzCD methylation. These findings suggested that in the regulation of PE response, two pathways communicate with each other to mediate adaptation. Here we provided evidence to indicate that DifA does not undergo methylation during EPS regulation and PE chemotaxis. On the other hand, using mutants expressing the NarX-DifA chimera, it was found that signal transduction through DifA, DifC (CheW-like) and DifE (CheA-like) modulates FrzCD methylation. Surprisingly, the attractant 12:0 PE can modulate FrzCD methylation in two ways distinguishable by the dependency on DifA, DifC and DifE. The DifACE-independent mechanism, which may result from specific sensing of 12:0 PE by FrzCD, increases FrzCD methylation as expected. Unexpectedly, 12:0 PE decreases FrzCD methylation with the DifACE-dependent mechanism. This "opposite" FrzCD methylation by DifACE-dependent signaling was supported by results from NafA-expressing mutants because nitrate, which acts as a repellent, increases FrzCD methylation. Based on these findings, we proposed a model for chemotaxis toward 12:0 PE (and 16:1 PE). In this model, DifA and FrzCD both sense the same signal and activate the pathways of excitation (Dif) and adaptation (Frz) independently. The two pathways communicate with each other via methylation crosstalk between DifACE and FrzCD in such a way that processes of excitation and adaptation can be coordinated. / Ph. D.

Biological signal processing can help us gain knowledge about biological complexity, as well as using this knowledge to engineer better systems. Three areas are identified as critical to understanding biology: 1) understanding DNA, 2) examining the overall biological function and 3) evaluating these systems in environmental (ie: turbulent) conditions. DNA is investigated for coding structure and redundancy, and a new tandem repeat region, an indicator of a neurodegenerative disease, is discovered. The linear algebraic framework can be used for further analysis and techniques. The work illustrates how signal processing is a tool to reverse engineer biological systems, and how our better understanding of biology can improve engineering designs. Then, the way a single-cell mobilizes in response to a chemical gradient, known as chemotaxis, is examined. Inspiration from receptor clustering in chemotaxis combined with a Hebbian learning method is shown to improve a gradient-source (chemical/thermal) localization algorithm. The algorithm is implemented, and its performance is evaluated in diffusive and turbulent environments. We then show that sensor cross-correlation can be used in solving chemical localization in difficult turbulent scenarios. This leads into future techniques which can be designed for gradient source tracking. These techniques pave the way for use of biologically-inspired sensor networks in chemical localization.

Résumé : Le sepsis sévère et le choc septique réfractaire sont d’incidences grandissantes dans la population et restent actuellement un défi de taille avec une mortalité variant de 30 à 70 % à 28 jours malgré l’amélioration du traitement de support. Les corticostéroïdes (CS) sont un traitement d’appoint controversé dans le sepsis sévère et le choc septique réfractaire. Ils modulent entre autre les fonctions des neutrophiles ou polymorphonuclear neutrophils cells (PMN) qui sont des acteurs de 1ère ligne dans la défense immédiate contre le sepsis et les défaillances organiques associées. Résultats : Le sepsis a pour effet de diminuer l’activité phagocytaire, ainsi que la production de radicaux libres oxygénés (ROS) des PMN au jour 1 (J1) et jour 3 (J3). Il augmente l’adhésion qui s’intensifie avec la sévérité de la maladie et persiste jusqu’à J3. Au niveau de la dégranulation, le sepsis augmente la production et la libération de la pentraxine 3 (PTX3). Le sepsis affecte le profil phénotypique des PMN en augmentant l’expression de CD66b et ST2 à J1 et J3. Il accentue également l’expression de CD64. Dans le groupe 2 (G2), ce niveau d’expression diminue à J3. Aucun effet significatif sur le chimiotactisme n’a été observé à J1, ni J3. L’usage de CS in vitro retarde l’apoptose à J1 et J3 dans le groupe 1 (G1), mais ne démontre aucune amélioration significative des fonctions des PMN ou au niveau de leur profil phénotypique. Conclusion : Le sepsis entraîne une immunoparalysie des PMN au niveau de leur migration et des fonctions effectrices. Non seulement cette paralysie augmente avec la sévérité de la maladie, mais elle persiste également après 3 jours suivant l’admission. De faibles doses de CS in vitro et in vivo dans le traitement du choc septique n’ont pas d’effet déterminant sur les PMN dans l’amélioration du pronostic des patients. Davantage de recherches seront nécessaires afin d’approfondir notre compréhension de l’impact d’un traitement aux CS sur les fonctions neutrophiliques dans un contexte septique. Ces derniers permettraient non seulement de mieux cibler leur utilisation dans le but d’arriver à un rapport bénéfique/risque avantageux dans le choc septique, mais aussi pour d’autres maladies inflammatoires. / Abstract : With the increasing rates of severe sepsis and refractory septic shock, there is still a challenge in mortality rates between 30 and 70 % at 28 days despite improved supportive care. Corticosteroids (CS) are a controversial supportive treatment in severe sepsis and refractory septic shock. They modulate the functions of PMN that are players in first line of immediate defence against the sepsis and associated to multiorgan failures. Results : Sepsis leads to a reduced phagocytic activity and ROS production at day 1 (D1) and day 3 (D3). It enhances the adhesion which increases with the severity of sepsis and persists until D3. In terms of degranulation, the sepsis increases the production and release of PTX3. Sepsis affects the phenotypic profile of PMN that increases the expressions of CD66b and ST2 at D1 and D3. It increases the CD64 expression but decreased at D3 in G2. No significant effect on chemotaxis was observed in D1, neither in D3. In G1, use of CS in vitro further delays apoptosis at D1 and D3, but is not showing any improvement in functions of PMN or in phenotypic profile. Conclusions : Sepsis induces an immune paralysis of PMN in their migration and effectives functions. Not only this paralysis increases with the severity of the sepsis, but it also persists after 3 days following the admission. Low doses of CS in vitro and in vivo in the treatment of septic shock have not determinant effect on PMN in improvement of the outcome of patients. More research is needed to learn more about the impacts of CS treatment on PMN functions in sepsis. This would contribute not only to ensure a better target ing of their use in order to achieve an advantageous benefits/risks ratio in septic shock, but also for others inflammatory diseases.

The engraftment of haemopoietic stem/progenitor cells (HSPCs) from umbilical cord blood (UCB) into adult recipients, although advantageous in terms of sourcing units, the decreased need to match donor and recipient and reduced risk of graft versus host disease (GvHD), is delayed compared to grafts using HSPCs from mobilised peripheral blood (MPB) or bone marrow (BM). One reason for this is the limited number of HSPCs (CD34+/CD133+ cells) in a unit of UCB compared to MPB or BM. The CXCR4-CXCL12 axis is widely recognised as a key player in the bone marrow homing, retention, and engraftment of HSPCs. The aim of this thesis was to investigate whether the engraftment of HSPCs from UCB into the bone marrow could be improved. Firstly, a novel in vitro 3D time-lapse chemotaxis assay to assess the homing capacity of human UCB CD133+ HSPCs, towards the chemokine CXCL12 was developed. One advantage of this assay was that it distinguished cell chemotaxis from chemokinesis and allowed these parameters to be quantified. Human UCB CD133+ HSPC chemotaxis towards CXCL12 was inhibited by the CXCR4 antagonist, AMD3100. Importantly, the presence of CXCL12 or AMD3100 had no affect on cell chemokinesis. To complement the in vitro chemotaxis assay, a short term in vivo homing assay in NSG mice was successfully established. The effect of siRNA silencing of the CXCR4 co-receptor, CD164, which is also expressed on CD133+ HSPCs, on cell migratory and homing ability was investigated. CD164 knock-down using siRNA in human UCB CD133+ HSPCs did not demonstrate an effect on homing to NSG bone marrow in vivo or chemotaxis to CXCL12 in vitro. However, homing to NSG mouse spleen was significantly reduced in cells silenced for CD164. Following this, an 8 day HSPC expansion system using nanofibre scaffolds (Nanex) and differing cytokines was investigated. These serum and feeder free conditions yielded a significant expansion of cells that retained CD133+CD34+ expression and their in vitro chemotactic ability to CXCL12. Time constraints did not permit the engrafting ability of these cells to be analysed in an in vivo HSC reconstitution assay that was initiated. However these studies will provide the basis to support future related research in this laboratory.

Chemotaxis, the phenomenon through which cells respond to external chemical signals, is one of the most important and universally observable in nature. It has been the object of considerable modelling effort in the last decades. The models for chemotaxis available in the literature cannot reconcile the dynamics of external chemical signals and the intracellular signalling pathways leading to the response of the cells. The reason is that models used for cells do not contain the distinction between the extracellular and intracellular domains. The work presented in this dissertation intends to resolve this issue. We set up a numerical hybrid simulation framework containing such description and enabling the coupling of models for phenomena occurring at extracellular and intracellular levels. Mathematically, this is achieved by the use of the fictitious domain method for finite elements, allowing the simulation of partial differential equations on evolving domains. In order to make the modelling of the membrane binding of chemical signals possible, we derive a suitable fictitious domain method for Robin boundary elliptic problems. We also display ways to minimise the computational cost of such simulation by deriving a suitable preconditioner for the linear systems resulting from the Robin fictitious domain method, as well as an efficient algorithm to compute fictitious domain specific linear operators. Lastly, we discuss the use of a simpler cell model from the literature and match it with our own model. Our numerical experiments show the relevance of the matching, as well as the stability and accuracy of the numerical scheme presented in the thesis.

The discovery of the prokaryotic cytoskeleton has revolutionized our thinking about spatial organisation in prokaryotes. However, the roles different bacterial cytoskeletal proteins play in the localisations of diverse biomolecules are controversial. Bacterial chemotaxis depends on signalling through large protein clusters and each cell must inherit a cluster on cytokinesis. In Escherichia coli the membrane chemosensory clusters are polar and new static clusters form at pre-cytokinetic sites, ensuring positioning at new poles after cytokinesis and suggesting a role for the bacterial FtsZ and MreB cytoskeletons. Rhodobacter sphaeroides has both polar, membrane-associated and cytoplasmic, chromosome-associated chemosensory clusters. This study sought to investigate the roles of FtsZ and MreB in the partitioning of the two chemosensory clusters in R. sphaeroides. The relative positioning between the two chemosensory systems, FtsZ and MreB in R. sphaeroides cells during the cell cycle was monitored using fluorescence microscopy. FtsZ forms polar spots after cytokinesis, which redistribute to the midcell forming nodes from which gradients of FtsZ extend circumferentially to form the Z-ring. The proposed node-precursor model might represent a common mechanism for the formation of cytokinetic rings. The MreB cytoskeleton continuously reorganizes between patchy and filamentous structures, and colocalises with FtsZ at midcell. Membrane chemosensory proteins form individual dynamic unit-clusters with mature clusters containing about 1000 CheW₃ proteins. These unit-clusters diffuse randomly within the membrane but have a higher propensity for curved regions like cell poles. Membrane clusters do not colocalise with FtsZ and MreB and appear excluded from the Z-ring vicinity. The bipolar localisation of membrane clusters is established after cell division via random diffusion and polar trapping of clusters. The cytoplasmic chemosensory clusters colocalise with FtsZ at midcell in new-born cells. Before cytokinesis one cluster moves to a daughter cell, followed by the second moving to the other cell. FtsZ and MreB do not participate in the positioning of cytoplasmic clusters. Therefore the two homologous chemosensory clusters use different mechanisms to ensure partitioning, and neither system utilizes FtsZ or MreB for positioning.

Neutrophil activation is the most important initial immune defense against invading microbes in newborn infants. The reduced neutrophil migration and uncontrolled regulation of reactive oxygen species (ROS) production observed in neonates, could result in a diminished infectious response or in tissue damage. The aims were to study neutrophil chemotactic response towards IL-8 and fMLP in term neonates; to examine neutrophil receptor expression involved in adhesion, migration, phagocytosis and complement after stimulation with IL-8 and fMLP in term neonates; and to investigate neutrophil production of ROS, induced by PMA and E.coli, after preincubation with IL-8 and fMLP in term and preterm newborn infants. Comparisons were made to neutrophils from healthy adults. Chemotaxis was distinguished from randomly migrating neutrophils, and the neutrophil migration distance and the number of migrating neutrophils per distance was evaluated. Neutrophils were labeled with antibodies to cell surface antigens (CD11b, CD18, CD65, CD15S, CD162, CD44, CD35, CD88, CD181, CD182 and CD64) after stimulation with IL-8 and fMLP. After preincubation of neutrophils with fMLP or IL-8 and stimulation with PMA or E.coli, respiratory burst was detected. The same analyses were also made in preterm infants (median 25+3weeks GA; range 23+0–29+2) within 3 days postnatal age. Neutrophils from neonates exhibited different migratory and receptor responses to IL-8 and fMLP, with a diminished response towards IL-8 in term newborn infants in terms of reduced chemotaxis and modulation of receptors involved in adhesion, chemotaxis, complement and phagocytosis as compared to adults. fMLP reduced PMA- and E.coli-induced respiratory burst in neutrophils from term neonates and adults. The reduced respiratory burst by fMLP may be a mechanism for reducing the detrimental effects of uncontrolled inflammation. Although a similar burst reduction was observed in preterm infants born >25 weeks GA with fMLP, a diminished neutrophil respiratory burst modulation in very preterm infants cannot be excluded and requires further studies at different gestational and postnatal ages.

From the developing embryo to the evacuation of football stadiums, the migration and movement of populations of individuals is a vital part of human life. Such movement often occurs in crowded conditions, where the space occupied by each individual impacts on the freedom of others. This thesis aims to analyse and understand the effects of occupied volume (volume exclusion) on the movement of the individual and the population. We consider, as a motivating system, the rearrangement of individuals required to turn a clump of cells into a functioning embryo. Specifically, we consider the migration of cranial neural crest cells in the developing chick embryo. Working closely with experimental collaborators we construct a hybrid model of the system, consisting of a continuum chemoattractant and individual-based cell description and find that multiple cell phenotypes are required for successful migration. In the crowded environment of the migratory system, volume exclusion is highly important and significantly enhances the speed of cell migration in our model, whilst reducing the numbers of individuals that can enter the domain. The developed model is used to make experimental predictions, that are tested in vivo, using cycles of modelling and experimental work to give greater insight into the biological system. Our formulated model is computational, and is thus difficult to analyse whilst considering different parameter regimes. The second part of the thesis is driven by the wish to systematically analyse our model. As such, it concentrates on developing new techniques to derive continuum equations from diffusive and chemotactic individual-based and hybrid models in one and two spatial dimensions with the incorporation of volume exclusion. We demonstrate the accuracy of our techniques under different parameter regimes and using different mechanisms of movement. In particular, we show that our derived continuum equations almost always compare better to data averaged over multiple simulations than the equivalent equations without volume exclusion. Thus we establish that volume exclusion has a substantial effect on the evolution of a migrating population.

Serpentes do gênero Bothrops são responsáveis por 90% dos acidentes ofídicos no Brasil. Seus venenos provocam efeitos locais em humanos e animais, como hemorragia, edema, dor e necrose, caracterizando uma resposta inflamatória, cujo mecanismo não está bem definido. Esses efeitos estão relacionados com a ação combinada de proteases, substâncias que induzem hemorragia e fosfolipases, bem como a liberação de mediadores endógenos gerados pelos venenos. Considerando que a ativação do sistema complemento (SC) e de funções celulares, como quimiotaxia, ativação, proliferação e citotoxicidade podem desempenhar papel importante nos processos inflamatórios e de lesão tecidual subsequentes ao envenenamento, o estudo propõe: a) investigar a capacidade dos venenos brutos de serpentes Bothrops jararacussu e Bothrops pirajai e das toxinas purificadas, serinoprotease de B. jararacussu (SPBj) e L-aminoácido oxidase de B. pirajai (LAAOBp), em modular a atividade do SC; b) avaliar a contribuição do efeito sobre o SC no recrutamento de leucócitos polimorfonucleares humanos (PMN); c) avaliar o potencial citotóxico direto dos venenos e toxinas sobre células mononucleares do sangue periférico humano (PBMC); d) analisar o efeito dos venenos sobre a modulação da expressão dos marcadores de ativação CD69, CD25 e HLA-DR em células T, B e natural killer (NK). Os resultados do ensaio de citotoxicidade mostraram que o veneno bruto de B. jararacussu foi citotóxico para PBMC apenas nas concentrações maiores, de 50 e 100g/mL, não apresentando citotoxicidade nas outras concentrações testadas. A serinoprotease apresentou baixa citotoxicidade para essas células, o que sugere a necessidade de maiores investigações quanto aos mecanismos que levam a essa morte celular. O aumento da viabilidade celular encontrado nas amostras incubadas com veneno bruto e LAAO de B. pirajai sugere possível indução de proliferação celular, que necessita de maiores estudos. Os resultados obtidos sugerem que os venenos brutos de B. jararacussu e B. pirajai são capazes de ativar o SC como observado nos ensaios cinéticos da VCVL e VA e de quimiotaxia de neutrófilos, onde ficou evidenciado que a migração celular foi devida a liberação dos fatores quimiotáticos do SC, C3a e C5a. e que suas respectivas toxinas, serinoprotease e LAAO apresentam efeitos moduladores sobre o SC humano, e estimulam investigações mais aprofundadas com a finalidade de se esclarecer os mecanismos de ação e identificar os componentes responsáveis pelos efeitos observados. Houve expressão aumentada de CD69, CD25 e HLA-DR nas células T CD4+ e CD8+, especialmente quando incubadas com veneno bruto de B. jararacussu e LAAO de B. pirajai, o que reflete ativação da resposta imune celular, e pode sugerir que este tipo de resposta desempenhe papel relevante na indução e/ou controle dos processos imunopatológicos decorrentes de envenenamentos por B. jararacussu e B. pirajai. Esta investigação visa fornecer subsídios para a possível utilização das toxinas para fins terapêuticos e como ferramentas para investigação dos mecanismos envolvidos nos processos fisiopatológicos que ocorrem em decorrência de picadas e também em outras doenças de caráter inflamatório. / Snakes of the genus Bothrops are responsible for 90% of snakebites in Brazil. Their venoms cause local effects in humans and animals, such as hemorrhage, edema, pain and necrosis, characteristic of an inflammatory response. The mechanism is not well defined. These effects are related to the combined action of proteases, substances that induce bleeding and phospholipases, as well as release of endogenous mediators generated by the venoms. Considering that activation of the complement system (CS) and cellular functions such as chemotaxis, activation, proliferation and cytotoxicity, may play a role in inflammatory processes and tissue injury following envenomation, the study proposes: a) to investigate the ability of crude venom of B. jararacussu and B. pirajai and the purified toxins, serineprotease of B. jararacussu and L-amino acid oxidase (LAAO) of B pirajai in modulating the activity of the CS, b) to assess the contribution of the effect on CS in the recruitment of human polymorphonuclear leukocytes (PMN), c) to assess the direct cytotoxic potential of venoms and toxins on human peripheral blood mononuclear cells (PBMC), d) to analyse the effect of venoms on the modulation of the expression of activation markers CD69, CD25 and HLA-DR on T, B and natural killer (NK) cells. The results of cytotoxicity assay showed that the crude venom of B. jararacussu was cytotoxic to PBMC only at higher concentrations, 50 and 100g/mL, showing no cytotoxicity in the other concentrations. The serineprotease showed low cytotoxicity to the cells, suggesting the need for further investigations about the mechanisms that lead to this cell death. The increase in cell viability found in samples incubated with crude venom of B. pirajai and LAAO suggests the possibility of induction of cell proliferation, which needs further study. The results suggest that the crude venom of B. jararacussu and B. pirajai are capable of activating the CS as observed in kinetic assays of classical pathwaylectin pathway and alternative pathway and neutrophil chemotaxis assay, where it was shown that cell migration was due to release of CS chemotactic factors, C3a and C5a, and that their respective toxins, serineprotease and LAAO have modulatory effects on human CS, and stimulate further research in order to clarify the mechanisms of action and identify the components responsible for the observed effects. There was increased expression of CD69, CD25 and HLA-DR on CD4+ and CD8+, especially when incubated with crude venom of B. jararacussu and LAAO of B. pirajai. It reflects activation of cellular immune response and may suggest that this type of response play an important role in the induction and/or control of immunopathological processes arising from envenomation by B. jararacussu and B. pirajai. This research aims to provide subsidies to the possible use of the toxin for therapeutic purposes and as tools for investigating mechanisms involved in pathophysiological processes that occur as a result of snakebites and also in other diseases of inflammatory nature.

INTRODUÇÃO: Emulsões lipídicas parenterais (EL) contendo óleo de peixe podem modular favoravelmente a resposta inflamatória e manter ou promover a resposta imunológica, mas há dados insuficientes sobre seu impacto em funções de células da imunidade inata. OBJETIVO: Verificar o efeito da administração endovenosa de EL composta por mistura de triglicérides de cadeia média e óleos de soja, oliva e peixe sobre a migração e fagocitose de leucócitos de ratos, em comparação à EL composta por mistura física de triglicérides de cadeia média e cadeia longa - TCM/TCL, suplementada ou não com óleo de peixe (OP). MÉTODOS: Ratos (Lewis) isogênicos (n=40) foram submetidos à cateterização da veia jugular externa para acesso parenteral. Os animais foram randomizados em quatro grupos, de acordo com sua infusão endovenosa: grupo SMOF: EL contendo 30% de óleo de soja (TCL), 30% de TCM, 25% de óleo de oliva e 15% de OP; grupo TCM/TCL: EL contendo TCM e TCL (1:1 v/v); grupo TCM/TCL/OP: EL composta por TCM/TCL com adição de OP (8:2 v/v); grupo SF: solução fisiológica. Um grupo de animais sem cateterismo venoso também foi desenvolvido (CO-NC). No quinto dia de experimento e após injeção de carvão coloidal pela veia caudal, amostras de sangue e tecido (fígado, pulmão e baço) foram coletadas para análise quimiotática de neutrófilos (câmara de Boyden adaptada) e quantificação do número de macrófagos fagocitantes do carvão coloidal (imunohistoquímica). Os dados foram analisados por ANOVA e pós-teste de Tukey. RESULTADOS: SMOF não alterou a quimiotaxia e fagocitose nos leucócitos estudados. TCM/TCL e TCM/TCL/OP aumentaram o número de macrófagos fagocitantes do fígado e pulmão e somente TCM/TCL/OP apresentou aumento no número de macrófagos fagocitantes no baço (p<0,05). CONCLUSÕES: 1) Emulsões lipídicas, independente de sua composição, não influenciaram a quimiotaxia de neutrófilos; 2) Emulsão lipídica composta por mistura de triglicérides de cadeia média e óleos de soja, oliva e peixe apresentou efeito neutro sobre a quimiotaxia, migração espontânea de neutrófilos e recrutamento de monócitos no fígado, pulmão e baço; 3) Emulsão lipídica de mistura física de triglicérides de cadeia média e cadeia longa estimulou o recrutamento de monócitos, com aumento do número de macrófagos fagocitantes no fígado e pulmão; 4) Emulsão lipídica de mistura física de triglicérides de cadeia média e cadeia longa enriquecida com emulsão lipídica de óleo de peixe, estimulou o recrutamento de monócitos, com aumento do número de macrófagos fagocitantes no fígado, pulmão e baço. / RATIONALE: Parenteral lipid emulsions (LE) with fish oil could modulate inflammatory response and promote or maintain immunologic response, but there are insufficient data about the impact on innate immunity cells functions. AIM: To evaluate the effects of endovenous infusion of LE containing mixture of medium-chain triglycerides and soybean, olive and fish oils on leukocytes migration and phagocytosis in rats, compared to a physical mixture of medium and long-chain triglycerides - MCT/LCT LE supplemented or not with fish oil (FO). METHOD: Isogenic Lewis rats (n=40) were submitted to jugular vein catheterization for parenteral access. The animals were randomized in four groups, according to their infusion: group SMOF: LE containing 30% of soybean oil (LCT), 30% MCT, 25% olive oil and 15% fish oil; group MCT/LCT: LE containing MCT and LCT (1:1 v/v); group MCT/LCT/FO: MCT/LCT LE enriched with fish oil based LE (8:2 v/v); group SS: saline. A non-surgical control (CO-NS) was also performed. In the 5th experimental day and after colloidal carbon injection in tail vein, blood and tissue (liver, lung and spleen) samples were collected for chemotaxis assay (adapted Boyden chamber) and colloidal carbon phagocyting-macrophages quantification (immunohistochemistry). ANOVA and Tukey post test were performed. RESULTS: SMOF LE didn?t influence leukocytes chemotaxis and phagocytosis. MCT/LCT and MCT/LCT/FO LE increased liver and lung resident phagocyting-macrophages number (p<0.05) and only in MCT/LCT/FO group, spleen resident phagocyting-macrophages number was increased (p<0.05). CONCLUSION: 1) Lipid emulsion, independently of composition, has no influence on neutrophils chemotaxis; 2) Lipid emulsion with a mixture of medium-chain triglycerides, soybean, olive and fish oils has neutral effect on neutrophil chemotaxis, random migration and monocyte recruitment to the liver, lung and spleen; 3) Lipid emulsion with a physical mixture of medium and long-chain triglycerides has stimulatory effect on monocyte recruitment, with increase of phagocyting-macrophages number in liver and lung; 4) Lipid emulsion with a physical mixture of medium and longchain triglycerides supplemented with fish oil, has stimulatory effect on monocyte recruitment, with increase of phagocyting-macrophages number in liver, lung and spleen.