Pathogen Recognition and Innate Immunity

Living in an oxygenated environment has required the evolution of effective cellular strategies to detect and detoxify metabolites of molecular oxygen known as reactive oxygen species. Here we review evidence that the appropriate and inappropriate production of oxidants, together with the ability of organisms to respond to oxidative stress, is intricately connected to ageing and life span.

Oxidants, oxidative stress and the biology of ageing.

Background: Pulmonary endothelial injury is a critical process in the pathogenesis of acute lung injury (ALI) during sepsis Heat shock protein A12B (HSPA12B) is

/pdf/heat-shock-protein-a12b-protects-vascular-endothelial-cells-3qj3tex105.pdf

Heat Shock Protein A12B Protects Vascular Endothelial Cells Against Sepsis-Induced Acute Lung Injury in Mice

Diabetes-specific microvascular disease is a leading cause of blindness, renal failure and nerve damage, and diabetes-accelerated atherosclerosis leads to increased risk of myocardial infarction, stroke and limb amputation. Four main molecular mechanisms have been implicated in glucose-mediated vascular damage. All seem to reflect a single hyperglycaemia-induced process of overproduction of superoxide by the mitochondrial electron-transport chain. This integrating paradigm provides a new conceptual framework for future research and drug discovery.

http://www.chem.uwec.edu/Chem454/BiochemDiabetes.pdf

Biochemistry and molecular cell biology of diabetic complications

▪ Abstract The innate immune system is a universal and ancient form of host defense against infection. Innate immune recognition relies on a limited number of germline-encoded receptors. These receptors evolved to recognize conserved products of microbial metabolism produced by microbial pathogens, but not by the host. Recognition of these molecular structures allows the immune system to distinguish infectious nonself from noninfectious self. Toll-like receptors play a major role in pathogen recognition and initiation of inflammatory and immune responses. Stimulation of Toll-like receptors by microbial products leads to the activation of signaling pathways that result in the induction of antimicrobial genes and inflammatory cytokines. In addition, stimulation of Toll-like receptors triggers dendritic cell maturation and results in the induction of costimulatory molecules and increased antigen-presenting capacity. Thus, microbial recognition by Toll-like receptors helps to direct adaptive immune responses ...

/pdf/innate-immune-recognition-3eedeqyq83.pdf

Innate Immune Recognition

Lipopolysaccharides were isolated from dehydratedCampylobacter jejuni by combination of the phenol-chloroform-petroleum ether and phenol-water extraction techniques. Biochemical characterizations of lipopolysaccharide were performed on the two fractions of highest purity. Neutral sugar analyses detected galactose, glucose, trace amounts of mannose, and an unidentified deoxy-hexose. The primary amino sugars were galactosamine, glucosamine, and glucosamine-phosphate. Chemical analyses of other lipopolysaccharide components included phosphate, 3-deoxy-d-manno-octulosonic acid (KDO), and fatty acids. The predominant fatty acids were 3-hydroxytetradecanoic and hexadecanoic acids with lesser amounts of tetradecanoic acid. 3-Hydroxytetradecanoic acids were bound to lipid A by both amide and ester linkages.

Characterization of campylobacter jejuni lipopolysaccharide

Summary Lipopolysaccharide (LPS) preparations from Salmonella S-form bacteria contain, in addition to S-form LPS, variable amounts of R-form material, lacking the O-specific polysaccharide. In the present study, we investigated the R-form LPS present in 22 LPS preparations derived from different Salmonella S-form strains belonging to 8 serological groups. The R-form part of the LPSs was separated by SDS-polyacrylamide gel electrophoresis and its antigen specificity analyzed by subsequent immunobloting using Salmonella R-antisera (anti-Ra, -Rbl, -Rb2, -Rc). The results show that different R-determinants were present in one and the same LPS preparation. The pattern of reaction varied considerably among individual strains. This variation was also present among strains of the same serogroup. Approximately one half of the R-form LPSs exhibited a weak or no reaction with Ra antiserum. In contrast, almost all R-form LPSs showed a significant reaction with Rbl antiserum. The electrophoretic mobility of the R-form material was very similar, exhibiting only minor differences among the different LPS preparations. In most cases, migration was similar to that of authentic Salmonella Ra-LPS; in some cases, the migration was somewhat faster resembling that of Rb1-LPS.

Immunoblot analysis of the R-form lipopolysaccharide from Salmonella S forms.

The present invention relates to a method for producing self-renewing, non-transformed macrophages comprising: culturing a cell preparation obtained from an organ obtained from a mammal in culture medium to which granulocyte macrophage colony-stimulating factor (GM- CSF) has been added; thereby differentiating the cell population into self-renewing, non- transformed macrophages. The present invention further relates to macrophages obtainable by the method of the invention as well as their use in medicine or medical/pharmaceutical research. Furthermore, the present invention relates to a method of identifying a compound having a pharmacological, differentiating, proliferative or anti-proliferative, cytotoxic or transforming effect on the self-renewing, non-transformed macrophages of the invention as well as a method of identifying a compound capable of treating a disease selected from the group consisting of an inflammatory disease, infections, asthma, contact allergies as well as the side effects associated with virus vector gene-therapy.

Method for the generation of non-transformed macrophage cell lines

Thirty years ago in 1954, we summarized our knowledge then of the chemistry and biology of endotoxins (73). At that time, the phenol/water extraction procedure had been established (74), and the extracts from a number of bacterial strains studied chemically and biologically. These investigations had led to the identification of the extracted products as the 0 antigens and endotoxins of these bacteria, resembling products isolated earlier by Boivin and Mesrobeanu (2), Morgan et al. (44), Goebel et al. (15), Shear et al. (62), and others. It had been recognized that endotoxins are composed of a polysaccharide component linked covalently to a lipid component, and thus were chemically lipopolysaccharides. The lipid had been termed lipid A (73) and found to contain D - glucosamine, phosphate, and long-chain fatty acids in ester and amide linkages. It had been shown that the linkage of the polysaccharide to lipid A could be cleaved by hydrolysis with dilute acetic or hydrochloric acid, leading to free water-soluble polysaccharide and free water-insoluble lipid A. Neither free polysaccharide nor free insoluble lipid A expressed endotoxic activities. Nevertheless, it has been anticipated that lipid A was the biologically active part and that activity would result from the binding of lipid A to the polysaccharide component, the latter functioning as a hydrophilic carrier for lipid A.

Lipid A: Relationships of Chemical Structure and Biological Activity

Lipopolysaccharides (LPS) extracted from the supersusceptible strain Pseudomonas aeruginosa Z61 were compared with LPS from other strains with varying antimicrobial susceptibilities. The presence of 4-amino-4-deoxy-arabinose (4-AraN) in P. aeruginosa Z61 LPS was confirmed by gas-liquid chromatography/mass spectrometry (GLC-MS) and quantitated by high-performance liquid chromatography (HPLC). Z61 LPS (compared with wild-type strain PAO1) has reduced amounts of rhamnose and higher concentrations of hydroxy fatty acids, 4-AraN, and phosphates. 31P Nuclear magnetic resonance revealed that Z61 LPS phosphates are configured in monophosphates, phosphodiesters, pyrophosphomonoesters, and glycosidic pyrophosphodiester groups. The presence of 4-AraN in P. aeruginosa LPS did not correlate with antimicrobial resistance.

The occurrence of 4-amino-4-deoxy-arabinose in LPS of supersusceptible strain Pseudomonas aeruginosa Z61: noncorrelation with polymyxin resistance.

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