Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P43026 (lipopolysaccharide)
 62,215 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Bacterial endotoxin (lipopolysaccharide, LPS) induces coagulation of horseshoe crab hemolymph. Our previous studies had demonstrated that a hemolymph factor, designated factor B, was associated with the LPS-mediated activation of the Limulus clotting system [Ohki et al. (1980) FEBS Lett. 120, 318-321]. On further purification of factor B we found that an additional component, designated factor C, was required to generate factor B activity in the presence of LPS in order to activate the proclotting enzyme. To elucidate the role of factor C in the LPS-mediated reaction, factor C was isolated and characterized from the hemocyte lysate under sterile conditions. The preparation exhibited a single band on sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) in the absence of 2-mercaptoethanol, while two protein bands on SDS-PAGE were observed after reduction. Thus, factor C had a Mr of 123 000 consisting of a heavy chain of Mr = 80 000 and a light chain of Mr = 43 000. Factor C was converted to an activated form in the presence of LPS with a Mr = 123 000, designated factor C. Upon activation, cleavage of the light chain occurred resulting in the accumulation of two new fragments of Mr = 34000 and 8500 on reduced SDS-PAGE. A diisopropylfluorophosphate-sensitive active site was localized in the light chain (Mr = 34000) of factor C. The reconstitution experiments, using factor C, factor B, proclotting enzyme and LPS, demonstrated that all of these proteins are essential for the endotoxin-mediated coagulation system. On the basis of these results we propose that a cascade pathway of LPS-induced activation of the Limulus clotting system consists of three sequential activations of hemolymph serine protease zymogens.
 ...
PMID:Lipopolysaccharide-sensitive serine-protease zymogen (factor C) found in Limulus hemocytes. Isolation and characterization. 351 66

An intracellular clotting factor, factor B, which is closely associated with the hemolymph coagulation system of horseshoe crab (Tachypleus tridentatus), was purified and characterized. The purified preparation gave a single band (Mr = 64,000) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the absence of 2-mercaptoethanol, while three bands (Mr = 64,000, 40,000, and 25,000) were detected on SDS-PAGE after reduction. This preparation was converted by limulus clotting factor C to an activated form, factor B, with Mr = 56,000 consisting of a heavy chain (Mr = 32,000) and a light chain (Mr = 25,000) bridged by disulfide linkage(s). The factor B, which was produced separately by treating the partially purified factor B with factor C, was also purified. It gave a single band on unreduced SDS-PAGE and two bands on reduced SDS-PAGE. The purified factor B had Mr of 56,000 consisting of a heavy chain (Mr = 32,000) and a light chain (Mr = 25,000). These results indicated that the purified factor B zymogen is a mixture of single-chain and two-chain forms, both of which have the same molecular weight of 64,000, and that these two forms are converted to factor B by factor C. The diisopropyl phosphorofluoridate-sensitive site of factor B was found in the heavy chain. The reconstitution studies using purified factor C, factor B, proclotting enzyme and coagulogen in the presence of lipopolysaccharide indicated that factor B is an essential component to complete sequential activation of the limulus clotting system, and that it specifically activates proclotting enzyme to the active clotting enzyme.
 ...
PMID:Purification and properties of intracellular clotting factor, factor B, from horseshoe crab (Tachypleus tridentatus) hemocytes. 351 94

Twenty-one parasite-naive dogs were infected with 60,000 protoscolices of Echinococcus granulosus. Transformation of peripheral lymphocytes was investigated before and 29 days after the infection, immunoglobulin concentration and anti-hydatid fluid protein (HFP) titers in serum and feces before and at 35 days of infection, skin reactivity to HFP at 36 days, and characteristics of the parasites at 40 days. The infection caused a significant depression of the spontaneous, lipopolysaccharide-stimulated, and purified protein derivative-stimulated blastogenesis. Responses to phytohemagglutinin were unchanged and reactivity to concanavalin A was enhanced with the infection. Only the concentrations of IgG and IgA in the serum and IgA in the feces increased significantly after infection. Fifteen (71%) dogs produced significant serum titers of anti-HFP hemagglutinins but copro-antibodies were detectable in only 3 dogs at minimum titers. Titers were abolished by treatment with 2-mercaptoethanol. The serum of 11 (52%) dogs transferred passive cutaneous anaphylaxis to guinea pigs but none transferred skin reactivity to pups or rabbits. Five and 1 (but not 0.2) micrograms of HFP caused skin reactivity in 4 parasite-naive dogs. Nineteen (90.5%) infected dogs reacted significantly to skin inoculation of 0.2 microgram of HFP at 0.5 hours and 13 (62%) at 6 hours. The 7 dogs with the highest anti-HFP serum titers or the greatest skin reactivity at 6 hours had significantly less mature or fewer tissue parasites, respectively, than the 7 dogs with the smallest responses. Since there was evidence that the specific immunity was still developing at the time of the study, these results indicate that immunological diagnosis of, and artificial immunization against, canine echinococcosis are feasible.
 ...
PMID:Humoral immunity in the prepatent primary infection of dogs with Echinococcus granulosus. 352 Oct 67

The effects of 3-acetyldeoxynivalenol (3-AcDON) on mitogen-induced lymphocyte proliferation and antibody production were studied in male CD-1 mice exposed to 0, 2.5, 5 or 10 ppm 3-AcDON in the diet for 35 days. Mitogen-induced lymphocyte proliferation and T-cell-independent antibody responses to dinitrophenyl-ficoll or Escherichia coli were not altered by dietary exposure to 3-AcDON. The T-cell-dependent antibody response to sheep red blood cells was increased in the group fed 10 ppm 3-AcDON. In vitro, 3-AcDON inhibited lymphocyte proliferation in a dose-dependent manner. Inhibition was observed when the toxin was present during the first 8 hr in phytohaemagglutinin-stimulated cultures and during the first 24 hr in lipopolysaccharide-stimulated cultures. This suggests that 3-AcDON blocks an early step in lymphocyte activation. This inhibition was not restored by thiol reducing agents (dithiothreitol, L-cysteine or 2-mercaptoethanol). Similarly, the addition of lymphokines, including interleukin-1 or interleukin-2, did not alter the inhibitory effects of 3-AcDON. These results suggest that the in vitro effects of 3-AcDON may not reflect its in vivo immunotoxicity. However, 3-AcDON may serve as a chemical probe for examining the activation process of lymphocyte proliferation.
 ...
PMID:Immunological responsiveness of mouse spleen cells after in vivo or in vitro exposure to 3-acetyldeoxynivalenol. 360 79

The availability of use of mouse peritoneal lymphocytes as target cells for analyzing sister chromatid exchanges (SCE) upon exposure to a genotoxic drug, cyclophosphamide, was investigated using female ICR mice. Use of these cells overcame the difficulty in use of mouse lymphocyte cultures, recovering sufficient metaphase cells. The greatest advantage of use of peritoneal lymphocytes was that about 1-2 X 10(6) lymphocytes/mouse could easily be recovered from the peritoneal cavity in high purity. Their mitogenic responses were good when Escherichia coli lipopolysaccharide, in combination with 2-mercaptoethanol, was used as mitogens, but they were less when purified phytohemagglutinin was used. In the presence of lipopolysaccharide (60 micrograms/ml) and 2-mercaptoethanol (22-88 microM), the maximum incidence of second division metaphases (greater than 50%) and the highest mitotic index (greater than 4%) were observed 36-40 h after stimulation. Under these conditions, the base-line SCE showed the constant level. The range of intrastrain variations in the base-line SCE was 0.24-0.36/chromosome. The distribution histograms of SCE/chromosome did not fit a single Poisson model, suggesting that these cells are heterogeneous with respect to the base-line SCE. Single s.c. injections 1 h before harvest of doses of 0.75-3.0 mg of cyclophosphamide per kg evoked positive responses, and injections of over 0.375 mg/kg had linear dose-dependent effects. On harvest of cells for up to 192 h after the injection, the maximal induction of SCE attained 1 h after exposure was found to return time dependently to the control level at 192 h. After the initial rapid reduction in the cell number, cellular recovery, measured as the mitotic index and the number of peritoneal exudate cells recovered, returned to the control level within 48 h, without a significant increase thereafter. After maintaining cells under the liquid-holding experiment for various times in vitro following a single exposure to cyclophosphamide for 1 h in vivo, the reduction of their SCE and recovery of their mitotic index were more rapid than those of cells in the time-course experiment. These findings suggest that the association of the recruitment of less- and/or nondamaged cells from their precursors with reduction of the SCE is slight. Repair(s) and, to a lesser extent, selective loss of more damaged cells may be the main factors contributing to the early reduction response of the SCE frequency. The relations of these factors are discussed.
 ...
PMID:Mouse peritoneal lymphocytes, a new target for analyzing induction of sister chromatid exchanges on in vivo exposure to a genotoxic agent. 370 68

The in vivo mitogenic responses to lipopolysaccharide or concanavalin A by spleen cells of mice exposed to 20 ppm nitrogen dioxide (NO2) for 96 hr, were evaluated. [3H]Thymidine incorporation after addition of either mitogen, was significantly lower in spleen cells from acutely NO2-exposed mice (NO2 SC) than from control mice, although cell viability was not affected. T- and B-cell mitogenic responses were inhibited to the same extent by NO2 exposure. NO2 SC responses were protected by the thiol compounds 2-mercaptoethanol, L-cysteine, and selenomethionine. No restoration of mitogenic response was observed after treatment with reduced glutathione. Mechanisms accounting for this in vivo NO2 immune toxicity, are discussed in terms of oxidative injury.
 ...
PMID:In vivo nitrogen dioxide exposure depresses spleen cell in vitro mitogenic responses: effects of sulfur compounds. 380 31

The ability of lipopolysaccharide (LPS) to influence lymphocyte mitogen responsiveness to 2-mercaptoethanol (2-ME) was investigated. 2-ME was mitogenic for splenocytes only when fetal bovine serum lots containing elevated endotoxin levels were used to supplement (5%) the culture medium. 2-ME did not stimulate mitosis of nylon-wool-fractionated splenocytes (NWSC); however, 2-ME induced significant proliferation in the NWSC cell preparations when endotoxin-deficient culture medium was supplemented with doses of LPS which alone did not stimulate significant proliferation. The sIg+ and Thy-1-/Lyt-1- fractions of NWSC, which represented less than 5% of the cells in the 'T-cell preparations', were almost completely dependent on 2-ME for proliferation in response to LPS stimulation. These data indicate that nylon-wool-fractionated preparations contain splenic B cells that incorporate significant amounts of [3H]thymidine when stimulated with LPS and 2-ME.
 ...
PMID:2-Mercaptoethanol-dependent lipopolysaccharide-responsive B cells in nylon-wool-fractionated spleen cell preparations. 387 36

The complement-requiring passive hemolysis test with Salmonella typhimurium lipopolysaccharide-coated sheep erythrocytes is more sensitive for antibodies directed against the lipopolysaccharide than is the passive hemagglutination test. The hemagglutinating and hemolyzing antibodies produced in Swiss mice by hyperimmunization, either with or without Freund's adjuvant, were distributed in both the light and heavy fractions isolated by sucrose density gradient fractionation and gel filtration. IgM fractions, whether tested by hemagglutination or hemolysis, were sensitive to 2-mercaptoethanol (0.15 m). On the other hand, IgG hemolytic antibodies were more sensitive to 2-mercaptoethanol than were IgG hemagglutinating antibodies. The resistance of IgG hemagglutinating activity amounted to about 72 to 95% of the total IgG recovered, whereas the resistant portion of the IgG hemolytic activity was approximately 40 to 53%. It is suggested that, although mercaptoethanol sensitivity is not a definitive test for IgM antibody, its use in connection with the hemagglutination test gives at least an approximation of the IgG antibody, whereas the hemolysis test gives a better approximation of maximal measurable antibody against Salmonella lipopolysaccharides.
 ...
PMID:Heterogeneity of antibody response to Salmonella lipopolysaccharide measured by passive hemagglutination and hemolysis in mice. 417 45

It has been proposed that two distinct signals are required for the triggering of the precursors of antibody-forming bone marrow-derived cells (B cells): (a) the binding of antigen or of a mitogen to the corresponding receptor sites on B-cell membranes and (b) the interaction of activated C3 with the C3 receptor of B lymphocytes. There is growing evidence that B-cell mitogens and T (thymus-derived cell)-independent antigens are capable of activating the alternate pathway of the complement system (bypass). Therefore, the effect of another potent bypass inducer was investigated with regard to B-cell activation and the role of C3. Purified, pyrogen-free cobra venom factor was mitogenic for both T and B lymphocytes (cortisone-resistant mouse thymus cells and lymph node lymphocytes from congenitally athymic mice). Venom factor could substitute for T cells by restoring the potential of antibody formation to sheep red blood cells in mouse B-cell cultures supplemented with macrophages or 2-mercaptoethanol. Venom factor may be capable of conferring activated C3 to the C3 receptor of B lymphocytes: preincubation of lymphoid cells with homologous serum or plasma, 10 mM EDTA, and sepharose-coupled venom factor converted with serum to an enzyme active against C3, inhibited their capacity to subsequently form rosettes with sheep erythrocytes sensitized with amboceptor and C5-deficient mouse complement. In the absence of EDTA, preincubation of freshly prepared B-cell suspensions with C3-sufficient homologous serum also blocked their subsequent interaction with complement-sensitized erythrocytes and at the same time rendered them reactive to an otherwise T-cell-specific mitogen. Moreover, mitogen induced B-cell proliferation in lymph node (but not in spleen) cell cultures, appeared to depend on the availability of exogenous C3: zymosan-absorbed fetal bovine serum (only 8.3% site-forming units remaining) supported T-cell activation by phytohemagglutinin, concanavalin A, and venom factor, but failed to sustain B-cell stimulation by pokeweed mitogen, lipopolysaccharide, and venom factor. T-cell-dependent antibody formation in composite cultures containing T cells or T-cell-substituting B-cell mitogens, B cells, and macrophages, always required the presence of C3-sufficient serum.
 ...
PMID:Complement-dependent B-cell activation by cobra venom factor and other mitogens? 458 89

Colicin D-CA23, obtained by sonic treatment of mitomycin C-induced cells of Escherichia coli K-12 W1485 (colD), was purified by ammonium sulfate precipitation, gel filtration on Sephadex G200, ion-exchange chromatography on diethylaminoethyl cellulose, and isoelectrofocusing. Polyacrylamide-gel electrophoresis, sedimentation velocity analysis, and antigenic analysis indicated that the preparation was homogeneous. Colicin D is composed entirely of amino acids and hence is a simple protein uncomplexed with lipid or lipopolysaccharide. It contains six residues of cysteine per molecule. The molecular weight of colicin D is approximately 92,000, as determined by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis and gel filtration on Sephadex G200. Its sedimentation coefficient is 4.41S. The behavior of colicin D in solutions of sodium dodecyl sulfate and 2-mercaptoethanol indicates that it does not consist of subunits and exists as a single polypeptide chain. Its high molecular weight and presence of six cysteine residues per molecule distinguish colicin D from all colicins previously described. Although colicins D and E3 have similar modes of action, their gross molecular properties are entirely different.
 ...
PMID:Purification and characterization of colicin D. 462 24


<< Previous 1 2 3 4 5 6 7 8 Next >>