Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UMLS:C0043167 (pertussis)
 19,595 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Nitric oxide (NO) induction was studied in the peritoneal macrophages and spleen cells of female NIH mice immunised with whole cell Bordetella pertussis vaccines of moderate and high potency, respectively. Compared with controls receiving diluent only, the macrophages and spleen cells of the vaccinated mice developed high levels of reactive nitrogen intermediates from the third day after injection. The nitrite concentrations achieved maximum values at the 10th day, but significant levels persisted until the 25th day. Heat-killed B. pertussis cells were the most effective inducer of NO synthesis, followed by lipopolysaccharide and agglutinogens Fim 2 and 3. Pertussis toxoid, filamentous haemagglutinin and pertactin were poor inducers of NO synthesis. The specific nitric oxide synthase inhibitor, aminoguanidine, and anti-IFN-gamma antibody blocked formation of nitrite by the macrophages and spleen cells. The production of NO in response to in vitro culture with bacterial antigen was clearly associated with protective immunity in vivo as determined by i.c. challenge. These results suggest that reactive nitrogen intermediates play a role in the immune response induced by whole cell pertussis vaccines.
 ...
PMID:Nitric oxide induction in murine macrophages and spleen cells by whole-cell Bordetella pertussis vaccine. 960 4

In disadvantaged populations, recurrent infections lead to a loss of body nitrogen and worsen nutritional status. The resulting malnutrition, in its turn, produces a greater susceptibility to infection. This study aimed to examine the ability of a new minimally invasive tracer protocol to measure leucine oxidation, and then to use it to quantify the effect of vaccination on leucine kinetics and oxidation. Undernourished men (n = 5; body mass index 16.3 +/- 0.9 kg/m(2)) and children (n = 9; age 4.1 +/- 0.6 y; weight-for-age Z-score -2.3 +/- 0.7) underwent metabolic studies 6 d before and 1 d after vaccination with diphtheria, pertussis and tetanus (DPT). The tracer protocol was performed in the fed state and involved two 3-h sequential periods of frequent (20 min) oral doses of NaH(13)CO(3) or [1-(13)C] leucine. Frequent breath samples and urine collections were made. Blood samples were obtained from the men and used for the determination of the isotopic enrichment of alpha-ketoisocaproic acid. The prevaccination oxidation of leucine (percentage of dose +/- SD) was 18.1 +/- 2.3 (men) and 16.7 +/- 3.8 (children). One day after vaccination, these values had risen to 19. 9 +/- 1.9 (P < 0.05) in the men and to 19.5 +/- 4.6 (P < 0.01) in the children. In the adults, vaccination was associated with a rise in whole-body protein breakdown [mg protein/(kg.h)] from 200 +/- 40 to 240 +/- 10 (P < 0.05). A minor simulated infection increases leucine catabolism in undernourished humans and this new, minimally invasive protocol is sufficiently sensitive to measure these changes.
 ...
PMID:A minimally invasive tracer protocol is effective for assessing the response of leucine kinetics and oxidation to vaccination in chronically energy-deficient adult males and children. 1041 87

In current Bordetella pertussis media ammonium accumulates because of an imbalance in the nitrogen:carbon ratio of the substrates used, which is one of the factors limiting cell density in fed-batch cultures. The aim of this study was to map B. pertussis catabolic and anabolic capabilities, in order to design a medium that avoids ammonium accumulation, while substrates are metabolised completely. Besides the known dysfunctional glycolysis, B. pertussis also possessed a partially dysfunctional citric-acid cycle. Although ammonium accumulation was avoided by adding various carbon sources to medium with glutamate, nuclear magnetic resonance (NMR) showed excretion of acetate, acetoacetate and beta-hydroxy-butyrate, thereby reducing the biomass yield. Acetoacetate and beta-hydroxy-butyrate were also formed in Verwey, B2 and modified Stainer-Scholte medium. Electron microscopy in combination with NMR showed that cells early on in these cultures contained poly-hydroxy-butyrate (PHB) globules, which disappeared later during the culture, coinciding with the appearance of beta-hydroxy-butyrate and/or acetoacetate. No globules nor metabolite excretion was detected when lactate in combination with glutamate were used as substrates. Thus, metabolite excretion and ammonium accumulation were avoided, while the yield of 8.8 g C-mol-1 compared favourably with literature values, averaging 6.5 g C-mol-1. Optimisation of this medium for pertussis toxin production will be reported in a separate article.
 ...
PMID:Rational medium design for Bordetella pertussis: basic metabolism. 1055 54

The intracerebral mouse protection test (Kendrick test) for the potency assay of pertussis vaccines is a complex and time consuming in vivo test which has a significant intra- and interlaboratory variation. Thus, there is a pressing need to develop a replacement for the Kendrick test. There is now convincing evidence to suggest that Bordetella pertussis can be taken up and survive within macrophages in the lungs and that cell-mediated immunity plays a role in protection. It was hypothesised that murine macrophages could be activated by immunisation with whole cell pertussis vaccines and therefore induce NO production. An alternative in vitro assay based on the determination of reactive nitrogen intermediates produced as a result of macrophage activation has been examined as a possible replacement for the current intracerebral (i.c.) mouse protection test. NO induction was studied in the peritoneal macrophages of female NIH mice immunised with normal and denatured whole cell B. pertussis vaccines respectively. Compared with controls receiving diluent only, macrophages and spleen cells from mice immunised with whole cell pertussis vaccine responded in vitro to selected pertussis antigens by NO synthesis. The production of NO in response to in vitro culture with bacterial antigen was immunisation dose dependent and was correlated with protective immunity in vivo as determined by i.c. challenge. The results suggest that NO production may serve as a marker of macrophage activation in mice immunised with whole cell vaccine, and could form the basis of a potential replacement potency assay.
 ...
PMID:Development of a nitric oxide induction assay as a potential replacement for the intracerebral mouse protection test for potency assay of pertussis whole cell vaccines. 1056 81

A single dose of lithium was injected intravenously or intraperitoneally in rats. Lithium levels in serum and tissues 5 or 24 hours later were elevated when the rats were pretreated with pertussis vaccine (PV). The vaccine was effective whether given locally (subcutaneous) or systemically (intravenous). Tests of heated (inactivated) PV suggested that pertussis toxin might be responsible for the effects of PV. Injection of purified pertussis toxin (PT) confirmed this suggestion. Elevation of serum urea nitrogen suggested that lithium levels were increased because the combination of PV or PT with lithium reduced renal excretory function which could cause retention of lithium. Inasmuch as PV and PT are known to inactivate the inhibitory G-proteins, these data suggest G-protein involvement in the elevation of lithium levels by PV and PT.
 ...
PMID:Pertussis vaccine and pertussis toxin increase lithium levels in rats: possible role of G-proteins. 1065 86

Inorganic polyphosphate (poly P) is a chain of tens or many hundreds of phosphate (Pi) residues linked by high-energy phosphoanhydride bonds. Despite inorganic polyphosphate's ubiquity--found in every cell in nature and likely conserved from prebiotic times--this polymer has been given scant attention. Among the reasons for this neglect of poly P have been the lack of sensitive, definitive, and facile analytical methods to assess its concentration in biological sources and the consequent lack of demonstrably important physiological functions. This review focuses on recent advances made possible by the introduction of novel, enzymatically based assays. The isolation and ready availability of Escherichia coli polyphosphate kinase (PPK) that can convert poly P and ADP to ATP and of a yeast exopolyphosphatase that can hydrolyze poly P to Pi, provide highly specific, sensitive, and facile assays adaptable to a high-throughput format. Beyond the reagents afforded by the use of these enzymes, their genes, when identified, mutated, and overexpressed, have offered insights into the physiological functions of poly P. Most notably, studies in E. coli reveal large accumulations of poly P in cellular responses to deficiencies in an amino acid, Pi, or nitrogen or to the stresses of a nutrient downshift or high salt. The ppk mutant, lacking PPK and thus severely deficient in poly P, also fails to express RpoS (a sigma factor for RNA polymerase), the regulatory protein that governs > or = 50 genes responsible for stationary-phase adaptations to resist starvation, heat and oxidant stresses, UV irradiation, etc. Most dramatically, ppk mutants die after only a few days in stationary phase. The high degree of homology of the PPK sequence in many bacteria, including some of the major pathogenic species (e.g. Mycobacterium tuberculosis, Neisseria meningitidis, Helicobacter pylori, Vibrio cholerae, Salmonella typhimurium, Shigella flexneri, Pseudomonas aeruginosa, Bordetella pertussis, and Yersinia pestis), has prompted the knockout of their ppk gene to determine the dependence of virulence on poly P and the potential of PPK as a target for antimicrobial drugs. In yeast and mammalian cells, exo- and endopolyphosphatases have been identified and isolated, but little is known about the synthesis of poly P or its physiologic functions. Whether microbe or human, all species depend on adaptations in the stationary phase, which is truly a dynamic phase of life. Most research is focused on the early and reproductive phases of organisms, which are rather brief intervals of rapid growth. More attention needs to be given to the extensive period of maturity. Survival of microbial species depends on being able to manage in the stationary phase. In view of the universality and complexity of basic biochemical mechanisms, it would be surprising if some of the variety of poly P functions observed in microorganisms did not apply to aspects of human growth and development, to aging, and to the aberrations of disease. Of theoretical interest regarding poly P is its antiquity in prebiotic evolution, which along with its high energy and phosphate content, make it a plausible precursor to RNA, DNA, and proteins. Practical interest in poly P includes many industrial applications, among which is the microbial removal of Pi in aquatic environments.
 ...
PMID:Inorganic polyphosphate: a molecule of many functions. 1087 45

The regulatory control of pertussis vaccines, as for other biological products, requires that they conform to specified standards of safety and efficacy. The current potency test for whole cell vaccines, the intracerebral mouse protection test (AMPT) is still the only such assay that has shown a correlation with protection in children. An alternative in vivo assay based on non-lethal aerosol challenge of mice has been assessed as a replacement for the current AMPT. An in vitro assay based on determination of reactive nitrogen/oxygen intermediates produced as a result of macrophage activation has also been investigated as a potential replacement for the in vivo challenge test. On the other hand, for safety testing, an enzymatic-HPLC coupled assay using a fluorescein-labelled G alpha(i3)C20 peptide to measure the enzymatic ribosylation activity of active pertussis toxin was evaluated for its suitability as a replacement for the current histamine sensitisation test (HIST). An assay for adenylate cyclase toxin (ACT)-related toxicity, based on measuring the ACT-induced oxidative burst in macrophage-like cell cultures has also been investigated. Although some questions still need to be answered in relation to the development of suitable replacements for in vivo tests of pertussis vaccines, the prospects for further improvements are promising.
 ...
PMID:Developments in reduction and replacement of in vivo toxicity and potency tests for pertussis vaccines. 1267 25

DPT, a combination vaccine against diphtheria, tetanus and pertussis is available since many years and still continued in the national immunisation schedule of many countries. Although highly potent, reactions to DPT vaccine are well known, mainly attributed to the factors like Pertussis component, aluminum adjuvant and lower purity of tetanus and diphtheria toxoids. The latter most important aspect has become a matter of concern, specially for the preparation of next generation combination vaccines with more number of antigens in combination with DPT. Purity of toxoid is expressed as Lf (Limes flocculation) per mg of protein nitrogen. The Kjeldahl method (KM) of protein nitrogen estimation suggested by WHO and British Pharmacopoeia is time consuming and less specific. Need has been felt to explore an alternative method which is quick and more specific for toxoid protein determination. DC (detergent compatible) protein assay, an improved Lowry's method, has been found to be much more advantageous than Kjeldahl method.
 ...
PMID:Comparative quantitation for the protein content of diphtheria and tetanus toxoids by DC protein assay and Kjeldahl method. 1293 7

A protocol for isolation of neutrophil plasma membranes utilizing a plasma membrane marker antibody, anti-CD15, attached to superparamagnetic beads was developed. Cells were initially disrupted by nitrogen cavitation and then incubated with anti-CD15 antibody-conjugated superparamagnetic beads. The beads were then washed to remove unbound cellular debris and cytosol. Recovered plasma membranes were quantified by immunodetection of G(beta2) in Western blots. This membrane marker-based separation yielded highly pure plasma membranes. This protocol has advantages over standard density sedimentation protocols for isolating plasma membrane in that it is faster and easily accommodates cell numbers as low as 10(6). These methods were coupled with immunodetection methods and an adenosine 5(')-diphosphate-ribosylation assay to measure the amount of membrane-associated G(ialpha) proteins available for receptor coupling in neutrophils either stimulated with N-formyl peptides or treated to differing degrees with pertussis toxin. As expected, pertussis toxin treatment decreased the amount of membrane G protein available for signaling although total membrane G protein was not affected. In addition, activation of neutrophils with N-formyl peptides resulted in an approximately 50% decrease in G protein associated with the plasma membrane.
 ...
PMID:Magnetic bead isolation of neutrophil plasma membranes and quantification of membrane-associated guanine nucleotide binding proteins. 1475 Dec 52

Whooping cough is a highly contagious respiratory disease caused by Bordetella pertussis Despite widespread vaccination, its incidence has been rising alarmingly, and yet, the physiology of B. pertussis remains poorly understood. We combined genome-scale metabolic reconstruction, a novel optimization algorithm, and experimental data to probe the full metabolic potential of this pathogen, using B. pertussis strain Tohama I as a reference. Experimental validation showed that B. pertussis secretes a significant proportion of nitrogen as arginine and purine nucleosides, which may contribute to modulation of the host response. We also found that B. pertussis can be unexpectedly versatile, being able to metabolize many compounds while displaying minimal nutrient requirements. It can grow without cysteine, using inorganic sulfur sources, such as thiosulfate, and it can grow on organic acids, such as citrate or lactate, as sole carbon sources, providing in vivo demonstration that its tricarboxylic acid (TCA) cycle is functional. Although the metabolic reconstruction of eight additional strains indicates that the structural genes underlying this metabolic flexibility are widespread, experimental validation suggests a role of strain-specific regulatory mechanisms in shaping metabolic capabilities. Among five alternative strains tested, three strains were shown to grow on substrate combinations requiring a functional TCA cycle, but only one strain could use thiosulfate. Finally, the metabolic model was used to rationally design growth media with >2-fold improvements in pertussis toxin production. This study thus provides novel insights into B. pertussis physiology and highlights the potential, but also the limitations, of models based solely on metabolic gene content.IMPORTANCE The metabolic capabilities of Bordetella pertussis, the causative agent of whooping cough, were investigated from a systems-level perspective. We constructed a comprehensive genome-scale metabolic model for B. pertussis and challenged its predictions experimentally. This systems approach shed light on new potential host-microbe interactions and allowed us to rationally design novel growth media with >2-fold improvements in pertussis toxin production. Most importantly, we also uncovered the potential for metabolic flexibility of B. pertussis (significantly larger range of substrates than previously alleged; novel active pathways allowing growth in minimal, nearly mineral nutrient combinations where only the carbon source must be organic), although our results also highlight the importance of strain-specific regulatory determinants in shaping metabolic capabilities. Deciphering the underlying regulatory mechanisms appears to be crucial for a comprehensive understanding of B. pertussis's lifestyle and the epidemiology of whooping cough. The contribution of metabolic models in this context will require the extension of the genome-scale metabolic model to integrate this regulatory dimension.
 ...
PMID:Probing the Genome-Scale Metabolic Landscape of Bordetella pertussis, the Causative Agent of Whooping Cough. 2884 44


<< Previous 1 2 3 4 Next >>