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Query: UNIPROT:P43026 (lipopolysaccharide)
 62,215 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Extracts of specific granules and azurophil granules from human neutrophils were tested for their bactericidal activity against various lipopolysaccharide mutants of Salmonella typhimurium LT-2. Three purified granule populations, one specific and two azurophil, were obtained by isopycnic centrifugation of homogenized neutrophils. Each was extracted with 0.2 M acetate buffer (pH 4), and the extracts were dialyzed against phosphate-buffered saline (pH 7) to remove acetate. These extracts contained >/=84% of the lysozyme, lactoferrin, or myeloperoxidase initially present in the whole granules. The S. typhimurium mutants possessed Ra, Rc, Rd(1), Rd(2), or Re lipopolysaccharide. As the carbohydrate content of the lipopolysaccharide decreased, the bacteria became increasingly more susceptible to the bactericidal activity of all granule extracts. Bactericidal activity of the extracts was in the order: mixed (azurophil + specific) >/= azurophil >> specific. Specific granules were bacteriostatic for S through Rd(2) bacteria. They were bactericidal only for the Re mutant. Both azurophil granule populations were equally bactericidal. Extracts boiled for 30 min retained none of their bactericidal activity for any of the bacteria; however, they remained bacteriostatic for the deep rough (Rd(2), Re) mutants. Bactericidal activity was dependent upon pH, in that mixed and azurophil granule contents killed the smooth parent and Ra mutant best at pH 5, the Rc and Rd(1) mutants to the same degree at pH 5 to 8, and the deep rough mutants (Rd(2) and Re) best at pH 8. Specific granule contents were most bacteriostatic for S through Rd(2) bacteria at pH 5 and killed the Re mutant only at pH 8. Thus, as the S. typhimurium lipopolysaccharide content decreased, the bactericidal pH optimum increased. Killing by all extracts was dependent upon incubation temperature, with almost no bactericidal or bacteriostatic activity observed when bacteria and granule fractions were incubated on ice (2 degrees C) and plated immediately. Intermediate killing was observed at 22 degrees C. If bacteria were incubated with granule extracts at 2 degrees C, washed free of extract, suspended in medium without extract, and reincubated at 37 degrees C, killing was observed. This suggested that a component(s) of the extracts was sticking to the bacteria at 2 degrees C but killing only at 37 degrees C.
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PMID:Bactericidal activity of specific and azurophil granules from human neutrophils: studies with outer-membrane mutants of Salmonella typhimurium LT-2. 2

Adaptations of the Farr technique have resulted in a specific and reproducible radioactive antigen-binding assay for antibodies directed against the lipopolysaccharide (LPS) of Neisseria meningitidis. LPS was intrinsically labeled with 14C acetate during 16-hr growth in a modified Frantz media, extracted by hot phenol-H2O, and purified by dialysis, ultracentrifugation, and ethanol precipitation. LPS, which aggregates in aqueous solutions, was maintained in a monomeric form in 3% sodium deoxycholate (NaD) as determined by gel filtration on Sephadex G-75. Since NaD is insoluble in (NH4)2SO4, polyethylene glycol, 20%, was used to precipitate immunoglobulins of all three major classes.
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PMID:Response to antigenic determinants of Neisseria meningitidis lipopolysaccharide investigated with a new radioactive antigen-binding assay. 5 1

The sugar composition of lipopolysaccharide (LPS) isolated from whole cells of Alteromonas haloplanktis 214 (previously referred to as marine pseudomonas B-16, ATCC 19855), variant 3, of the lipid A, core, and side-chain fractions derived from it, and of the LPS fractions (LPS I, II, and III) obtained by subjecting it to preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis has been determined. Conditions optimum for the release of constituent monosaccharides by hydrolysis were established. Sugars were quantitated by gas-liquid chromatography of their alditol acetate derivatives. Lipid A was detected by gel electrophoresis and by the spectral shift obtained with a carbocyanin dye. A comparison of the molar ratios of the various fractions suggest that LPS III is an LPS molecule lacking an O-antigenic side chain, whereas LPS I and II are LPS molecules differing in side-chain composition. LPS I may be a mixture of two LPS species. In double immunodiffusion experiments using anti-whole-cell serum, LPS I and II showed a homologous cross-reaction with isolated whole-cell LPS. LPS III as well as lipid A, core, and side-chain fractions failed to give rise to precipitin lines.
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PMID:Composition of the fractions separated by polyacrylamide gel electrophoresis of the lipopolysaccharide of a marine bacterium. 10 10

Membrane vesicles were prepared from Azotobacter vinelandii spheroplasts by lysis in either potassium phosphate (pH 7.0) or Tris1-acetate (pH 7.8) buffers. These 2 types of preparations differ considerably in their properties: 1) Examination by scanning electron microscopy reveals that the Pi vesicles consist primarily of closed structures 0.6-0.8 micrometer in diameter with a rough or particulate surface similar to that of spheroplasts. The Tris vesicles are significantly smaller, 0.1-0.3 micrometer in diameter, and have a much smoother surface structure. 2) Antisera from rabbits immunized with A. vinelandii lipopolysaccharide antigen will agglutinate Pi vesicles but not Tris vesicles. 3) Tris vesicles have a fourfold higher specific activity of latent H+-ATPase than Pi vesicles. After exposure to Triton X-100 similar ATPase activities are observed for both types of vesicles. 4) Pi vesicles transport calcium in the presence of ATP or lactate at less than 30% of the rats observed for Tris vesicles. 5) Tris vesicles have less than 22% of the transport capacity of Pi vesicles for accumulation of labeled sucrose and less than 3% of the capacity for valinomycin-induced uptake of rubidium observed during respiration. 6) Quinacrine fluorescence intensity is reduced by 30% during lactate oxidation and 20% during ATP hydrolysis by Tris vesicles. Under similar conditions, fluorescence in Pi vesicles is quenched by only 7% and less than 2%, respectively. These findings suggest that Pi vesicles have the normal orientation of the intact cell whereas Tris vesicles have an inverted topology.
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PMID:Isolation of membrane vesicles with inverted topology by osmotic lysis of Azotobacter vinelandii spheroplasts. 14 14

12-O-Tetradecanoylphorbol-13-acetate, a potent promoter of carcinogenesis in mouse skin, enhanced differentiation of cultured mouse myeloid leukemia cells (M1) induced by human urinary protein or by lipopolysaccharide from Salmonella typhosa. 12-O-Tetradecanoylphorbol-13-acetate enhanced differentiation of all the markers tested, such as phagocytosis, Fc rosette formation, lysozyme activity, and morphological change. Other potent tumor-promoting macrocyclic plant diterpenes also enhanced the induction of differentiation, but no-tumor-promoting diterpenes did not. These findings were in marked contrast with generally accepted findings on the inhibitory effect of 12-O-tetradecanoylphorbol-13-acetate on terminal differentiation observed in other cell culture systems but consistent with the observations with some kinds of leukemia cells.
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PMID:Enhancing effect of phorbol esters on induction of differentiation of mouse myeloid leukemia cells by human urinary protein and lipopolysaccharide. 29 79

CBA mice recieved a single intraperitoneal injection of hydrocortisone acetate (OHC) in a dose of 125 mg/kg body weight. At various times therafter, electrophoretic mobility (EPM), surface immunoglobulin (SIG) and in vitro DNA synthetic reactivity to concanavalin A (ConA), phytohemagglutinin (PHA), lipopolysaccharide (LPS) and tuberculin (PPD) were investigated on splenic lymphocytes. OHC was found to deplete rapidly the spleen to a minimum of 18% of control cellularity by day 4 posttreatment. At this time, the proportions of low mobility (LM) and SIG-bearing lymphocytes (B cells) were reduced respectively to 28% (control 54%) and 20% (control 45%). The proportion of high mobility (HM) lymphocytes (T cells) was increased to 72% (control 45%). While the mean EPM of LM cells (0.71) was only slightly and transiently reduced, that of HM cells was significantly augmented (1.24) over control value (1.16). This latter finding was interpreted as indicating the selective removal by OHC of a T cell subpopulation with a mean EPM around 1.10. Changes in mitogenic responsiveness were consistent with these alterations of B and T cell compartments. Despite a marked drop in spontaneous 3H-thymidine uptake, the absolute response to T cell mitogens ConA and PHA remained relatively unchanged. By contrast, the reactivity to B cell mitogens LPS and PPD was strongly depressed. Starting by day 12, regeneration and normalizaiton of lymphocyte populations proceeded slowly and were not achieved before day 26-34.
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PMID:Electrokinetic properties and mitogen responsiveness of mouse splenic B and T lymphocytes following hydrocortisone treatment. 30 Jul 13

Peripheral lymphocytes responsive to stimulation by phytohaemagglutinin (PHA) and bacterial lipopolysaccharide (LPS) in culture have been quantitated following treatment of mice with anti-thymocyte serum (ATS), total body irradiation or corticosteroids. The ATS reduced the number of PHA-responsive cells in both blood and spleen, and induced splenomegaly, but it had little deleterious effect on spleen-borne LPS-responsive cells. In contrast, the spleens of mice treated with hydrocortisone acetate were atrophied and the remaining cells had a reduced LPS response and an enhanced PHA response. Total body irradiation impaired both PHA and LPS responsiveness in the spleen. Recovery of PHA responsiveness after either irradiation or ATS treatment was prolonged and was dependent on the presence of an intact thymus; recovery of LPS responsiveness after corticosteroid treatment was more rapid and was thymus-independent.
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PMID:Effects of irradiation, anti-thymocyte serum and corticosteroids on PHA and LPS responsive cells of the mouse. 32 22

The effects of a variety of agents on the mouse antibody response to sheep red cells (SRC) and Escherichia coli lipopolysaccharide (LPS) were investigated. Agents formed two broad groups according to the time-dependence of their effects. Class I agents inhibited responses if given at any time over a broad period from before administration of antigen to 1-2 days afterwards. They included ionizing radiation, cortisone acetate, cholera toxin and several alkylating agents. Class II agents inhibited responses substantially only if given 1-2 days after the antigen. These agents were all either antimetabolites or vinca alkaloids. Class II agents had only moderate selectivity. When given at the time of maximal effectiveness they usually suppressed the response to SRC more than that to LPS. Some class I agents had striking selectivity. Radiation, cortisone acetate, and cholera toxin markedly suppressed the SRC response but had much less effect on the LPS response. In contrast, mitoclomine and its analogues, which are lipid-soluble alkylating agents, suppressed the response to LPS far more than that to SRC. Possible reasons for these differences in time-dependence and selectivity are discussed.
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PMID:Time-dependence and selectivity of immunosuppressive agents. 37 50

Proteins from human polymorphonuclear leukocyte granules were extracted with 0.2 M acetate, pH 4.0, and fractionated by Sephadex G-100 column chromatography. The fractions demonstrated selective bactericidal action against a deep rough cell wall mutant of Escherichia coli O111:B4 with rough lipopolysacharide and cell wall mutants of Salmonella typhimurium LT-2 with lipoplysacharide of Ra, Rc, Rd1, Rd2, and Re types. Smooth parent strains were most resistant to the bactericidal action. Fractions with greatest activity for the mutants were from valley regions (regions of low protein concentration) between three high protein peaks comprising myeloperoxidase, protease, and lysozyme, respectively. Susceptibility of the mutants to bactericidal action increased as sugar residues decreased in lipopolysaccharide. Gram-positive bacteria were susceptible to different fractions than were the gram-negative bacteria.
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PMID:Bactericidal activity of fractionated granule contents from human polymorphonuclear leukocytes. 37 30

Granule extracts from human polymorphonuclear leukocytes (PMN) were prepared with 0.2 M (pH 4.0) acetate. A fraction (valley AB) with distinctive bactericidal activity against cell wall mutants of Salmonella typhimurium LT-2 was obtained after fractionation of the granule extracts by Sephadex G-100 column chromatography. The smooth parent LT-2 strain was less sensitive to the bactericidal action. Susceptibility of the rough mutants to bactericidal action increased as sugar residues decreased in the lipopolysaccharide (LPS) (Re greater than Rd2 greater than Rd1 greater than Rc greater than Ra). Cationic protein(s) responsible for bactericidal activity could be selectively removed from the fraction by absorption with whole LT-2 cells or purified LPS. Loss of cationic protein species was confirmed by cationic polyacrylamide gel electrophoresis. Purified LPS from LT-2 or the deep rough mutant TA2168 inhibited the antimicrobial activity of the killing fraction in in vitro assays. A minor protein species (vAB1) from the valley AB fraction had an apparent molecular weight of 36,000 to 37,000 and represented a major bactericidal activity of the fraction. Small amounts of the isolated vAB1 protein were bactericidal for the smooth parent LT-2 strain.
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PMID:Bactericidal activity of fractionated granule contents from human polymorphonuclear leukocytes: antagonism of granule cationic proteins by lipopolysaccharide. 38 1


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