lipid-a and Burns

Endotoxin as the inciting agent of cytokines and other mediators, whose high level expression correlates with the septic shock and MOF, has been the one of leading causes of death in ICU.. For treating sepsis and MOF caused by endotoxin, the anti-lipid A of LPS antibody was used, 19 burned patients whose TBSA varied from 50% to 100% were divided into anti-LPS treatment group and nontreated group.. The levels of serum endotoxin, IL-6, IL-8, TNF and soluble IL-2R were lower obviously in patients of anti-LPS group than those of nontreated group (P < 0.05).. Clinical study surggests that anti-lipid A of LPS antibody can act as an therapeutic agent against gram-negative bacterin infection in burned patients.

Topics: Adult; Antibodies; Antitoxins; Burns; Cytokines; Endotoxemia; Endotoxins; Female; Humans; Interleukin-6; Interleukin-8; Lipid A; Male; Tumor Necrosis Factor-alpha

Monophosphoryl lipid A (MPLA) is a TLR4 agonist that has potent immunomodulatory properties and modulates innate immune function to improve host resistance to infection with common nosocomial pathogens in mice. The goal of this study was to assess the safety and efficacy of MPLA in a sheep model of burn injury and Pseudomonas aeruginosa pneumonia. The sheep provides a favorable model for preclinical testing as their response to TLR4 agonists closely mimics that of humans.. Twelve chronically instrumented adult female Merino sheep received 20% total body surface area, third-degree cutaneous burn under anesthesia and analgesia. At 24 h after burn, sheep were randomly allocated to receive: MPLA (2.5 μg/kg i.v., n = 6), or vehicle (i.v., n = 6). At 24 h after MPLA or vehicle treatment, Pseudomonas aeruginosa pneumonia was induced. Sheep were mechanically ventilated, fluid resuscitated and cardiopulmonary variables were monitored for 24 h after induction of pneumonia. Cytokine production, vascular barrier function, and lung bacterial burden were also measured.. MPLA infusion induced small and transient alterations in core body temperature, heart rate, pulmonary artery pressure, and pulmonary vascular resistance. Pulmonary mechanics were not altered. Vehicle-treated sheep developed severe acute lung injury during Pseudomonas aeruginosa pneumonia, which was attenuated by MPLA as indicated by improved PaO2/FiO2 ratio, oxygenation index, and shunt fraction. Sheep treated with MPLA also exhibited less vascular leak, lower blood lactate levels, and lower modified organ injury score. MPLA treatment attenuated systemic cytokine production and decreased lung bacterial burden.. MPLA was well tolerated in burned sheep and attenuated development of acute lung injury, lactatemia, cytokinemia, vascular leak, and hemodynamic changes caused by Pseudomonas aeruginosa pneumonia.

Topics: Adjuvants, Immunologic; Animals; Burns; Disease Models, Animal; Female; Lipid A; Multiple Organ Failure; Pneumonia, Bacterial; Pseudomonas aeruginosa; Pseudomonas Infections; Sheep

Infection is the leading cause of death in severely burned patients that survive the acute phase of injury. Neutrophils are the first line of defense against infections, but hospitalized burn patients frequently cannot mount an appropriate innate response to infection. Thus, immune therapeutic approaches aimed at improving neutrophil functions after burn injury may be beneficial. Prophylactic treatment with the TLR4 agonist monophosphoryl lipid A is known to augment resistance to infection by enhancing neutrophil recruitment and facilitating bacterial clearance. This study aimed to define mechanisms by which monophosphoryl lipid A treatment improves bacterial clearance and survival in a model of burn-wound sepsis. Burn-injured mice were treated with monophosphoryl lipid A or vehicle, and neutrophil mobilization was evaluated in the presence or absence of Pseudomonas aeruginosa infection. Monophosphoryl lipid A treatment induced significant mobilization of neutrophils from the bone marrow into the blood and sites of infection. Neutrophil mobilization was associated with decreased bone marrow neutrophil CXCR4 expression and increased plasma G-CSF concentrations. Neutralization of G-CSF before monophosphoryl lipid A administration blocked monophosphoryl lipid A-induced expansion of bone marrow myeloid progenitors and mobilization of neutrophils into the blood and their recruitment to the site of infection. G-CSF neutralization ablated the enhanced bacterial clearance and survival benefit endowed by monophosphoryl lipid A in burn-wound-infected mice. Our findings provide convincing evidence that monophosphoryl lipid A-induced G-CSF facilitates early expansion, mobilization, and recruitment of neutrophils to the site of infection after burn injury, allowing for a robust immune response to infection.

Topics: Animals; Burns; Granulocyte Colony-Stimulating Factor; Lipid A; Mice; Mice, Inbred BALB C; Neutrophil Infiltration; Neutrophils; Pseudomonas aeruginosa; Pseudomonas Infections; Receptors, CXCR4

Monophosphoryl lipid A (MPLA) is a Toll-like receptor 4 (TLR4) agonist that is currently used as a vaccine adjuvant in humans. In this study, we evaluated the effect of MPLA treatment on the innate immune response to systemic bacterial infections in mice. Mice treated with MPLA after burn injury showed improved survival and less local and systemic dissemination of bacteria in a model of Pseudomonas aeruginosa burn wound infection. Prophylactic treatment with MPLA significantly enhanced bacterial clearance at the site of infection and reduced systemic dissemination of bacteria despite causing attenuation of proinflammatory cytokine production during acute intra-abdominal infection caused by cecal ligation and puncture. Administration of MPLA at 1 h after CLP also improved bacterial clearance but did not alter cytokine production. MPLA treatment increased the numbers of granulocytes, double-positive myeloid cells, and macrophages at sites of infection and increased the percentage and total numbers of myeloid cells mediating phagocytosis of bacteria. Depletion of Ly6G(+) neutrophils, but not macrophages, eliminated the ability of MPLA treatment to improve bacterial clearance. The immunomodulatory effects of MPLA were absent in TLR4-deficient mice. In conclusion, these studies show that MPLA treatment significantly augments the innate immune response to bacterial infection by enhancing bacterial clearance despite the attenuation of proinflammatory cytokine production. The enhanced bacterial clearance is mediated, in part, by increased numbers of myeloid cells with effective phagocytic functions at sites of infection and is TLR4 dependent.

Topics: Animals; Burns; Cytokines; Female; Immunity, Innate; Lipid A; Lymphocyte Count; Macrophages; Male; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Neutrophils; Pseudomonas aeruginosa; Pseudomonas Infections; Signal Transduction; Toll-Like Receptor 4; Wound Infection

C57BL/6 mice were subjected to a full thickness scald thermal injury covering 25% of their total body surface area, and thioglycollate elicited peritoneal macrophages (Mphi were isolated 4 days later. Mphi from injured mice produced significantly greater amounts of reactive nitrogen intermediates and tumor necrosis factor-alpha in response to lipopolysaccharide and lipid A. Pertussis toxin (PTX) treatment of Mphi dose-dependently inhibited reactive nitrogen intermediate production in Mphi from sham-treated mice; however, Mphi from injured mice were insensitive to PTX-mediated inhibition. Conversely, tumor necrosis factor-alpha production was enhanced by PTX treatment, with Mphi from injured mice being more sensitive than Mphi from sham-treated mice to this effect of PTX. These results indicate that thermal injury increases Mphi sensitivity to lipopolysaccharide by a mechanism that is both PTX sensitive and PTX insensitive, thereby suggesting a role for G proteins in the modulation of Mphi activity after thermal injury.

Topics: Animals; Burns; Kinetics; Lipid A; Lipopolysaccharides; Macrophage Activation; Macrophages, Peritoneal; Mice; Mice, Inbred C57BL; Nitrogen Oxides; Pertussis Toxin; Reference Values; Thioglycolates; Tumor Necrosis Factor-alpha; Virulence Factors, Bordetella

A lipid A monoclonal antibody(mAb) was prepared and it was used to study its protective effect against burn sepsis. Wistar rats were inflicted with 30% TBSA third degress burn, and they were given LPS to mimic early sepsis after burn. The rats were divided randomly into burn with LPS, monoclonal antibody treatment, and control groups. The levels of endotoxin, tumor necrosis factor, light and electron microscopic studies of the morphological changes in the liver were studied. The results showed that the anti-lipid A monoclonal antibody demonstrated capacity to cross-react with several Gram-negative bacteria and their endotoxins. The mAb improved the survival rate of rats and decreased the levels of endotoxin and TNF as well as the liver damage significantly.

Topics: Animals; Antibodies, Monoclonal; Burns; Female; Lipid A; Male; Mice; Mice, Inbred BALB C; Random Allocation; Rats; Rats, Wistar; Sepsis

The role of lipopolysaccharide (LPS) in the virulence of Pseudomonas aeruginosa was studied. The virulence of several P. aeruginosa strains for burned mice was found to be directly related to the dispersion of LPS into either the phenol or the water phase after extraction. Virulence decreased as the proportion of LPS recovered from the phenol phase increased. No similar correlation was observed when several other strain characteristics were investigated. This phenomenon was studied in greater detail by using the "smooth"-specific phage E79 to select mutants altered in LPS structure. One such mutant, PA220-R2, was extensively characterized. LPS isolated from PA220-R2 was found to be completely deficient in high-molecular-weight polysaccharide material. This alteration rendered the strain serum sensitive and dramatically changed the reaction with O-specific typing sera and sensitivity to typing phages. However, motility, toxin A and elastase production, and 22 metabolic functions remained unchanged. PA220-R2 was found to be comparatively nonvirulent, with a 50% lethal dose more than 1,000-fold higher than that of its parent for burned mice. This was due to the inability of PA220-R2 to establish an infection in burned skin.

Topics: Animals; Burns; Endopeptidases; Galactosamine; Hexosamines; Lipid A; Lipopolysaccharides; Mice; Mutation; Pancreatic Elastase; Pseudomonas aeruginosa; Pseudomonas Infections; Sepsis; Serine Endopeptidases; Serotyping; Virulence