Decrease In Chemotaxis Research Articles

Oxi-1 is required for chemotaxis to odorants sensed by AWA but not AWC neurons.

This study examines the role of the oxi-1 UBE3B gene in chemotaxis of C. elegans to volatile odorants. Compared to wild type worms, oxi-1 mutants showed no difference in chemotaxis to the AWC-specific odorant, isoamyl alcohol but a significant decrease in chemotaxis compared to odr-7 mutants. Both oxi-1 and odr-7 mutants exhibited significant decreases in chemotaxis to AWA-specific odorants, pyrazine and diacetyl. For thiazole, which is sensed by both AWA and AWC neurons, only odr-7 mutants showed significantly decreased chemotaxis. These data demonstrate oxi-1 is required for chemotaxis to AWA- but not AWC-specific odorants, the mechanisms of which should be investigated.

Behavior of Chemokine Receptor 6 (CXCR6) in Complex with CXCL16 Soluble form Chemokine by Molecular Dynamic Simulations: General Protein‒Ligand Interaction Model and 3D-QSAR Studies of Synthetic Antagonists.

The CXCR6‒CXCL16 axis is involved in several pathological processes, and its overexpression has been detected in different types of cancer, such as prostate, breast, ovary, and lung cancer, along with schwannomas, in which it promotes invasion and metastasis. Moreover, this axis is involved in atherosclerosis, type 1 diabetes, primary immune thrombocytopenia, vitiligo, and other autoimmune diseases, in which it is responsible for the infiltration of different immune system cells. The 3D structure of CXCR6 and CXCL16 has not been experimentally resolved; therefore, homology modeling and molecular dynamics simulations could be useful for the study of this signaling axis. In this work, a homology model of CXCR6 and a soluble form of CXCL16 (CXCR6‒CXCL16s) are reported to study the interactions between CXCR6 and CXCL16s through coarse-grained molecular dynamics (CG-MD) simulations. CG-MD simulations showed the two activation steps of CXCR6 through a decrease in the distance between the chemokine and the transmembrane region (TM) of CXCR6 and transmembrane rotational changes and polar interactions between transmembrane segments. The polar interactions between TM3, TM5, and TM6 are fundamental to functional conformation and the meta-active state of CXCR6. The interactions between D77-R280 and T243-TM7 could be related to the functional conformation of CXCR6; alternatively, the interaction between Q195-Q244 and N248 could be related to an inactive state due to the loss of this interaction, and an arginine cage broken in the presence of CXCL16s allows the meta-active state of CXCR6. A general protein‒ligand interaction supports the relevance of TM3‒TM5‒TM6 interactions, presenting three relevant pharmacophoric features: HAc (H-bond acceptor), HDn (H-bond donator), and Hph (hydrophobic), distributed around the space between extracellular loops (ECLs) and TMs. The HDn feature is close to TM3 and TM6; likewise, the HAc and Hph features are close to ECL1 and ECL2 and could block the rotation and interactions between TM3‒TM6 and the interactions of CXCL16s with the ECLs. Tridimensional quantitative structure-activity relationships (3D-QSAR) models show that the positive steric (VdW) and electrostatic fields coincide with the steric and positive electrostatic region of the exo-azabicyclo[3.3.1]nonane scaffold in the best pIC50 ligands. This substructure is close to the E274 residue and therefore relevant to the activity of CXCR6. These data could help with the design of new molecules that inhibit chemokine binding or antagonize the receptor based on the activation mechanism of CXCR6 and provoke a decrease in chemotaxis caused by the CXCR6‒CXCL16 axis.

The Chemokine Receptors CXCR1 and CXCR2 Couple to Distinct G Protein-Coupled Receptor Kinases To Mediate and Regulate Leukocyte Functions

The chemokine receptors, CXCR1 and CXCR2, couple to Gαi to induce leukocyte recruitment and activation at sites of inflammation. Upon activation by CXCL8, these receptors become phosphorylated, desensitized, and internalized. In this study, we investigated the role of different G protein-coupled receptor kinases (GRKs) in CXCR1- and CXCR2-mediated cellular functions. To that end, short hairpin RNA was used to inhibit GRK2, 3, 5, and 6 in RBL-2H3 cells stably expressing CXCR1 or CXCR2, and CXCL8-mediated receptor activation and regulation were assessed. Inhibition of GRK2 and GRK6 increased CXCR1 and CXCR2 resistance to phosphorylation, desensitization, and internalization, respectively, and enhanced CXCL8-induced phosphoinositide hydrolysis and exocytosis in vitro. GRK2 depletion diminished CXCR1-induced ERK1/2 phosphorylation but had no effect on CXCR2-induced ERK1/2 phosphorylation. GRK6 depletion had no significant effect on CXCR1 function. However, peritoneal neutrophils from mice deficient in GRK6 (GRK6(-/-)) displayed an increase in CXCR2-mediated G protein activation but in vitro exhibited a decrease in chemotaxis, receptor desensitization, and internalization relative to wild-type (GRK6(+/+)) cells. In contrast, neutrophil recruitment in vivo in GRK6(-/-) mice was increased in response to delivery of CXCL1 through the air pouch model. In a wound-closure assay, GRK6(-/-) mice showed enhanced myeloperoxidase activity, suggesting enhanced neutrophil recruitment, and faster wound closure compared with GRK6(+/+) animals. Taken together, the results indicate that CXCR1 and CXCR2 couple to distinct GRK isoforms to mediate and regulate inflammatory responses. CXCR1 predominantly couples to GRK2, whereas CXCR2 interacts with GRK6 to negatively regulate receptor sensitization and trafficking, thus affecting cell signaling and angiogenesis.

Glycogen Synthase Kinase-3β regulates antigen/SCF-mediated cytokine production and SCF-mediated chemotaxis in human mast cells (93.6)

Abstract Glycogen Synthase Kinase-3β (GSK3β) is a serine threonine kinase downstream of PI3K, which has been implicated in cellular responses including tumorigenesis; cell growth, migration and survival; and cytokine generation. In this study we investigated the role of GSK3β in antigen/SCF-mediated mast cell responses. Short hairpin RNA (shRNA), introduced by lentiviral transfection, was used to knockdown GSK3β in human mast cells. Cytokine production and activation of transcription factors were measured by ELISA and immunoblot analysis. SCF-mediated mast cell chemotaxis was assessed with a transwell migration assay. GSK3β knockdown significantly decreased antigen/SCF-mediated GM-CSF and IL-8 production and SCF-mediated mast cell chemotaxis compared to control treated cells. The decrease in chemotaxis and cytokine production was associated with a pronounced reduction in antigen/SCF-mediated phosphorylation of JNK and the transcription factors c-jun and ATF2. These data are consistent with the conclusion that GSK3β is an essential component of the signalling cascade that regulates antigen/SCF-mediated transcriptional activity and chemotaxis in human mast cells. Thus, targeting GSK3β?may provide one means for down-regulation of these mast cell responses when associated with the allergic inflammatory reaction. Supported by the Division of Intramural Research, NIH, USA.

Omega-3 Fatty Acids for Maintenance of Remission of Crohn's Disease

Omega-3 Fatty Acids for Maintenance of Remission of Crohn's Disease

Decreased polymorphonuclear leucocyte chemotactic response to leukotriene B4 in cystic fibrosis.

Evidence that leukotriene B4 (LTB4) is a significant inflammatory mediator in chronic pseudomonal respiratory disease was sought in adolescents and young adults with cystic fibrosis. Specific chemotaxis of peripheral blood polymorphonuclear leucocytes (PMN) was used as an indirect measure of remote in vivo exposure to LTB4. PMN from 17 patients showed a significant decrease in chemotaxis to 10(-7)-10(-9) M LTB4, but normal responses to 10(-8) M n-formyl-methionyl-leucyl-phenylalanine and 4 mg/ml casein, when compared with 17 healthy age- and sex-matched controls. This result is consistent with chronic production of LTB4, and specific deactivation of circulating PMN receptors for LTB4 in patients with cystic fibrosis. Pharmacologic inhibition of LTB4 production in vivo may help elucidate its role in the pathogenesis of lung damage in cystic fibrosis.

Modulatory effect of ionizing radiation on food‐NaCl associative learning: the role of γ subunit of G protein inCaenorhabditis elegans

Ionizing radiation (IR) is known to impair learning by suppressing adult neurogenesis in the hippocampus. However, in a mature nervous system, IR-induced functional alterations that are independent of neurogenesis remain largely unknown. In the present study, we analyzed the effects of IR on a food-NaCl associative learning paradigm of adult Caenorhabditis elegans that does not undergo neurogenesis. We observed that a decrease in chemotaxis toward NaCl occurs only after combined starvation and exposure to NaCl. Exposure to IR induced an additional decrease in chemotaxis immediately after an acute dose in the transition stage of the food-NaCl associative learning. Strikingly, chronic irradiation induced negative chemotaxis in the exposed animals, i.e., the primary avoidance response. IR-induced additional decreases in chemotaxis after acute and chronic irradiation were significantly suppressed in the gpc-1 mutant, which was defective in GPC-1 (one of the two gamma subunits of the heterotrimeric G-protein). Chemotaxis to cAMP, but not to lysine and benzaldehyde, was influenced by IR during the food-NaCl associative learning. Our novel findings suggest that IR behaves as a modulator in the food-NaCl associative learning via C. elegans GPC-1 and a specific neuronal network and may shed light on the modulatory effect of IR on learning.

Bexarotene blunts malignant T‐cell chemotaxis in Sezary syndrome: Reduction of chemokine receptor 4‐positive lymphocytes and decreased chemotaxis to thymus and activation‐regulated chemokine

The malignant cells in Sezary syndrome express the skin trafficking molecules' cutaneous lymphocyte associated antigen (CLA) and chemokine receptor 4 (CCR4). High levels of the CCR4 ligand, thymus, and activation-regulated chemokine (TARC), have been reported in the blood and skin of patients. The rexinoid X-receptor specific retinoid, bexarotene, has contributed to the resolution of cutaneous disease among patients. To evaluate the effects of bexarotene on skin trafficking molecule expression and chemotaxis, peripheral blood mononuclear cells from Sezary syndrome patients and healthy controls were treated with bexarotene in vitro. CCR4 and CLA expression levels and chemotaxis in response to TARC (6.25 ng/ml) were evaluated among lymphocytes before and after treatment with bexarotene (10 microM). Flow cytometric analysis was performed to evaluate CD4, CD26, CLA, and CCR4 cell surface expression. Transwell migration assays were performed to evaluate chemotaxis to TARC. Prior to treatment, malignant cells exhibited higher CCR4 expression (45-90%) and greater than four times more chemotaxis to TARC compared with healthy controls. After treatment with bexarotene for 36-96 hr, a 28% reduction in CCR4 expression was noted (P < 0.05) among the malignant population with an associated 9% decrease in chemotaxis to TARC (P < 0.05). Our results show that bexarotene may inhibit malignant cell trafficking to the skin through an ability to suppress CCR4 expression among Sezary syndrome lymphocytes.

Effects of .GAMMA.-Ray Irradiation on Olfactory Adaptation to Benzaldehyde in Caenorhabditis elegans

Learning impairment following ionizing irradiation is an important potential risk for astronauts. Using Caenorhabditis elegans as a model organism for studying the nervous system, we previously showed that ionizing radiation affected salt chemotaxis learning behavior only at the transition stage of learning conditioning, which induced additional decreases in chemotaxis to NaCl immediately after irradiation. In the present study, we investigated the effects of γ-ray irradiation on olfactory adaptation to benzaldehyde, in which animals show a decrease in chemotaxis to benzaldehyde. Irradiation during the conditioning for adaptation to benzaldehyde did not induce an additional decrease in chemotaxis immediately after irradiation. On the other hand, at 1 h after irradiation, the progress of adaptation to benzaldehyde was interrupted by irradiation with the highest dose. These results indicate that impaired olfactory adaptation to benzaldehyde following irradiation is different from salt chemotaxis learning in C. elegans. Our findings suggest that the profile of radiation-induced response depends on the function of the nervous system in C. elegans.

Ascites from cirrhotic patients induces angiogenesis through the phosphoinositide 3-kinase/Akt signaling pathway

Ascites in patients with cirrhosis is associated with worsening of systemic hemodynamics. Thus, the aim of this study was to investigate the biological activity of ascites on endothelial cells. Human umbilical vein endothelial cells (HUVECs) were used to investigate the angiogenic activity of ascites obtained from cirrhotic patients. Ascites-induced Akt activation, cell migration and tube formation in HUVECs. The pretreatment of HUVECs with the phosphatidylinositide 3-kinase (PI3-kinase) inhibitor LY294002, resulted in a decrease in chemotaxis and cell tube formation induced by ascites. Moreover, the inhibition of Akt activity in HUVECs by transduction of an inactive phosphorylation Akt mutant (AA-Akt), blocked tube formation. These angiogenic effects of ascites were also operative in vivo showing a PI3-kinase activation dependence in the angiogenesis induced by ascites. In addition, the preincubation of ascites with anti-fibronectin antibody led to a significant decrease in HUVECs migration, cell tube formation and in vivo angiogenesis. These results confirm the novel concept that ascites is a bioactive fluid which can modify vascular properties through the activation of the PI3-kinase/Akt pathway in endothelial cells. Furthermore, our results demonstrated that this ascites-induced mechanism is mediated, at least in part, by fibronectin.

Activating and inhibitory Ly49 receptors modulate NK cell chemotaxis to CXC chemokine ligand (CXCL) 10 and CXCL12.

NK cells can migrate into sites of inflammatory responses or malignancies in response to chemokines. Target killing by rodent NK cells is restricted by opposing signals from inhibitory and activating Ly49 receptors. The rat NK leukemic cell line RNK16 constitutively expresses functional receptors for the inflammatory chemokine CXC chemokine ligand (CXCL)10 (CXCR3) and the homeostatic chemokine CXCL12 (CXCR4). RNK-16 cells transfected with either the activating Ly49D receptor or the inhibitory Ly49A receptor were used to examine the effects of NK receptor ligation on CXCL10- and CXCL12-mediated chemotaxis. Ligation of Ly49A, either with Abs or its MHC class I ligand H2-D(d), led to a decrease in chemotactic responses to either CXCL10 or CXCL12. In contrast, Ly49D ligation with Abs or H2-D(d) led to an increase in migration toward CXCL10, but a decrease in chemotaxis toward CXCL12. Ly49-dependent effects on RNK-16 chemotaxis were not the result of surface modulation of CXCR3 or CXCR4 as demonstrated by flow cytometry. A mutation of the Src homology phosphatase-1 binding motif in Ly49A completely abrogated Ly49-dependent effects on both CXCL10 and CXCL12 chemotaxis, suggesting a role for Src homology phosphatase-1 in Ly49A/chemokine receptor cross-talk. Ly49D-transfected cells were pretreated with the Syk kinase inhibitor Piceatannol before ligation, which abrogated the previously observed changes in migration toward CXCL10 and CXCL12. Piceatannol also abrogated Ly49A-dependent inhibition of chemotaxis toward CXCL10, but not CXCL12. Collectively, these data suggest that Ly49 receptors can influence NK cell chemotaxis within sites of inflammation or tumor growth upon interaction with target cells.

Regulation of macrophage function by the antioxidant N-acetylcysteine in mouse-oxidative stress by endotoxin

Changes in several functions of peritoneal macrophages from mice with oxidative stress caused by intraperitoneal injection of endotoxin ( Escherichia coli lipopolysaccharide, LPS) (100 mg/kg), and associated with a high production of reactive oxygen species (ROS), have been observed in our previous studies. Antioxidants such as N-acetylcysteine (NAC) are free radical scavengers that improve and modulate the immune response, especially in oxidative stress situations. Therefore, in the present work, we have studied the effects of the administration of NAC (150 mg/kg i.p.) on different functions of peritoneal macrophages from Swiss mice suffering that oxidative stress, caused by LPS (100 mg/kg). NAC was injected 30 min after LPS injection, and the peritoneal macrophages were obtained at 2, 4, 12, and 24 h after endotoxin injection. The following functions, key stages of the phagocytic process, were studied: adherence to substrate, chemotaxis, ingestion of particles, and production of ROS (reactive oxygen species), as well as tumor necrosis factor (TNFα) release. The decrease in chemotaxis and the increase in adherence, ingestion, superoxide anion production, and TNFα release shown by macrophages from animals with oxidative stress were counteracted by NAC injection. These data suggest that NAC administration may be useful for the treatment of oxidative stress-linked endotoxic shock, modulating the function of macrophages, specifically in decreasing the production of ROS and of inflammatory cytokines such as TNFα.

DIRECT CHEMOTACTIC EFFECT OF BRADYKININ AND RELATED PEPTIDES—SIGNIFICANCE OF AMINO- AND CARBOXYTERMINAL CHARACTER OF OLIGOPEPTIDES IN CHEMOTAXIS OF TETRAHYMENA PYRIFORMIS

The chemotactic character of the nonapeptide bradykinin (BK1–9) and its derivatives was studied in the eukaryotic ciliated model Tetrahymena pyriformis. The results demonstrate that BK1–9 has a direct and ligand-specific chemoattractant effect (maximal at 10−11m) without any intermediate substance as is essential in some mammalian test systems. Evaluation of the chemotactic effect elicited by derivatives showed that the presence of N- and C-terminal arginines can influence chemotactic potency of the molecule via expression of pyrrolidine and aromatic ring structures of terminal amino acid residues. Removal of the N-terminal Arg (expression of Pro) results in a significant decrease in chemotaxis (BK2–9), while further truncation of the C-terminal, causing expression of the aromatic ring of Phe (BK2–8), results in a highly chemoattractant variant. A single pyrrolidine ring on the C-terminus BK1–7 also has a positive effect on the chemotactic character, however further truncation (BK1–6, BK1–5) causes the chemoattractant character to become chemorepellent. Study of chemotactic selection with BK derivatives supports our previous findings that only phylogenetically selected ligands or their close derivatives are able to induce long-term selection with chemotaxis.

Plasticity of chemotaxis revealed by paired presentation of a chemoattractant and starvation in the nematode Caenorhabditis elegans.

While the basic functioning of the nervous system of Caenorhabditis elegans has been extensively studied, its behavioural plasticities have not been fully explored because of the limited availability of assay systems. We report here a simple form of chemotaxis plasticity in this organism: when worms are starved on plates that contain NaCl, their chemotaxis towards NaCl falls dramatically. This conditioning requires both the presence of NaCl and the absence of a bacterial food source, indicating that it is not merely adaptation or habituation, but that it is likely to be a form of associative learning. While chemotaxis towards volatile chemoattractants does not change significantly after conditioning with NaCl, chemotaxis towards other water-soluble attractants does decrease. This suggests that an altered response of a cell or a group of cells specifically involved in chemotaxis towards water-soluble chemoattractants is responsible for the behavioural alteration. The decrease in chemotaxis occurred slowly over 3-4 h of conditioning and returned quickly to the original level when either of the conditioning stimuli, NaCl or starvation, was removed. The application of serotonin partially blocked this reduction in chemotaxis, consistent with the proposed function of this neurotransmitter in food signalling. Using this assay, we have isolated three mutants with reduced plasticity. This assay system expands the opportunities for studying the molecular and cellular mechanisms of behavioural plasticity in C. elegans.

Leukocyte adhesion deficiency II-from A to almost Z.

Leukocyte adhesion deficiency (LAD) type II is the second human disorder identified which involves the adhesion cascade. While in LAD I the integrin family is defective, in LAD II the selectin system is involved. The syndrome has been described in only five patients and is transmitted as an autosomal recessive trait. The infectious episodes and the severity are much milder than those observed in LAD I, and the only persistent clinical symptom is chronic severe periodontitis. Delay separation of the umbilical cord, which is a hallmark for LAD I, was not observed in any of the LAD II patients. The exact defect in the system is absence of the SLeX, which is an important ligand for the selectin on the leukocyte lead ing to a profound defect in leukocyte rolling, the first step in the adhesion cascade. This causes a marked decrease in chemotaxis accompanied by pronounced neutrophilia. Apart from the leukocyte defect, these patients suffer from severe growth and mental retardation and exhibit the rare Bombay blood group type. The primary defect in the syndrome is in fucose metabolism, with the absence of all fucosylated glycans on cell surface membranes. Recently, it is was found that the defect is in a specific transporter of GDP fucose into the Golgi apparatus, and thus no fucosylation process takes place, and no surface expression can be detected. The exact genetic defect in the transporter is still unknown. Four of the patients were of Arabic origin while the fifth was of Turkish origin. It seems that the primary defect is somewhat different and, therefore, fucose administration was effective in the Turkish child, but did not show any beneficial results in the patients of Arabic origin.

Comparative study of peritoneal macrophage functions in mice receiving lethal and non-lethal doses of LPS

In previous studies, we have observed changes in several functions of peritoneal macrophages from female BALB/c mice with lethal endotoxic shock caused by intraperitoneal injection of Escherichia coli O55:B5 lipopolysaccharide (LPS; 100 mg/kg), which were associated with a high production of superoxide anion and tumor necrosis factor alpha (TNF-alpha). In the present work, both a lethal dose (250 mg/kg) and a non-lethal dose (100 mg/kg) of LPS were used in female Swiss mice. In peritoneal macrophages, the following functions were studied at 2, 4, 12 and 24 h after LPS injection: adherence to substrate, chemotaxis, ingestion of particles, and superoxide anion and TNF-alpha production. In both groups, the results showed a stimulation of adherence, ingestion and superoxide production as well as a decrease of chemotaxis, whereas TNF-alpha could not be detected in either of the two groups. These effects were more evident with the 250 mg/kg dose, especially as regards superoxide anion production, which was higher in the animals treated with a lethal dose of LPS.

Effect of Induced Molting on Heterophil Function in White Leghorn Hens

This study was undertaken to determine the effects of induced molt on basal functional activities of heterophils from aging hens. For this purpose, heterophils from both molted and unmolted hens were examined by in vitro bioassays for functional responsiveness and efficiency. We evaluated the ability of the heterophils to migrate to chemotactic stimuli, phagocytize opsonized and nonopsonized Salmonella-enteritidis (SE), and generate an oxidative burst in response to inflammatory agonists. A significant (P < 0.001) heterophilia was found in the molted hens within 2 days after feed withdrawal and remained throughout the length of the experimental feed withdrawal period. No significant differences were found in the random migration of heterophils from either group. The chemotactic movement of heterophils from molted hens was not affected until 8 days after feed withdrawal when compared with heterophil chemotaxis from unmolted hens. A significant decrease in chemotaxis by the heterophils from molted hens was observed days 8-12 after feed withdrawal (P < 0.05). Significantly (P < 0.05) fewer heterophils from molted hens were able to phagocytize opsonized (59% vs. 38%) and nonopsonized (26% vs. 15%) SE within 2 days after feed withdrawal. Likewise, significantly (P < 0.05) fewer bacteria were phagocytized per heterophil from the molted hens when compared with the number of bacteria per heterophil from the unmolted hens. The oxidative burst of heterophils stimulated by either opsonized zymosan A or phorbol myristate acetate of heterophils from molted hens was significantly (P < 0.05) reduced when compared with that generated by heterophils from the unmolted hens. These results indicate that feed withdrawal to induce molt alters the number and function of peripheral blood heterophils. This decreased efficiency of heterophil functional activity appears to play a role in the increased susceptibility of molting hens to SE infections.

Measurement of the Respiratory Burst and Chemotaxis in Polymorphonuclear Leukocs from Anti-Che Insecticides-Exposed Workers

Neutrophil function in 32 workers occupationally exposed to anti-ChE insecticides, as measured by chemotaxis through the leading front method and nitroblue tetrazolium dye reduction, was investigated and compared to those of age-and sex-matched controls. The cholinesterase (ChE) activity was normal in all the workers studied, although decrease of chemotaxis and of nitroblue tetrazolium reduction was observed in the exposed population. These results suggest that the identified functional changes in polymorphonuclear neutrophils might be an early indicator of anti-ChE insecticides toxicity, even in those individuals with no impairment in the ChE activity.

Effects of lead on neutrophil functions in occupationally exposed workers

Chemotactic and intracellular killing activity of neutrophils were examined in 25 male lead-exposed workers from storage-battery plants and compared to 25 healthy males with no history of lead exposure. Lead exposure was assessed using blood lead levels measured by atomic absorption spectrophotometry and zinc protoporphyrin levels assayed by hematofluorometry. Chemotaxis was carried out in Boyden chambers using zymosan activated serum as chemotactic stimulus. Intracellular killing activity of neutrophils was measured using nitroblue tetrazolium reduction test, measured of 515 nm in spectrophotometry. In lead-exposed workers a significant decrease in chemotaxis and random migration of neutrophils ( p<0.001) was observed compared to controls. Intracellular killing activity of polymorphonuclear leukocytes have also seemed to be slightly but not significantly reduced. These results suggest that human chronic exposure to lead may diminish neutrophil function in man.

Effects of endotoxic shock in several functions of murine peritoneal macrophages.

Gram negative sepsis and septic shock continue to be a major medical problem, with a complex physiopathology and it is associated with high mortality. Although secretion of cytokines such as tumor necrosis factor-alpha by macrophages is the principal host mediator of septic shock, other characteristic functions of macrophages implicated in their phagocytic capacity have not been studied in the process of endotoxic shock. In the present study we have used an intraperitoneal injection of E. coli lipopolysaccharide (LPS) (100 mg/kg) in order to obtain an endotoxic shock model in adult female BALB/c mice. Peritoneal cell suspensions were obtained at several times (2, 4, 12 or 24 h) after injection and the following functions were studied on the peritoneal macrophages: adherence to substrate, mobility (spontaneous and directed or chemotaxis), ingestion of particles and superoxide anion production. The results showed a stimulation of adherence, ingestion and superoxide production as well as a decrease of chemotaxis in the animals injected with LPS. These effects changed with time after LPS injection. Thus, the increase of adherence and the decrease of mobility were higher during the first hours, whereas the increase in ingestion and superoxide production turned larger with time.