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

Multiple carbohydrate structures on the outer-membrane lipopolysaccharide (LPS) of the gram-negative pathogen H. influenzae undergo high frequency, reversible loss, indicative of phase variation. Characterization of a genetic locus, lic-1, responsible for expression of two LPS epitopes displaying phase variation, showed it to comprise four genes. The first gene mediates phase variation. At its 5' end, within the open reading frame, are a variable number of tandem repeats of the tetramer CAAT. By shifting upstream initiation codons in or out of frame, these 4 bp units create a translational switch. The phenotype of organisms corresponds to the number of 4 bp units. Phase variation between three levels of expression ( +, +, and -) of lic-1-derived epitopes is caused by differences in the three phases of translation of the 5' terminus of this gene. Phase variation also allows for selection of organisms displaying certain LPS epitopes in vivo.
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PMID:The molecular mechanism of phase variation of H. influenzae lipopolysaccharide. 247 81

Outer membranes of Haemophilus influenzae type b were fractionated to yield Triton X-100-insoluble material and lipopolysaccharide and phospholipids. Liposomes reconstituted from lipopolysaccharide and phospholipids were impermeable to sucrose (Mr, 342) and to a high-molecular-weight dextran (average Mr, 6,600). When the Triton X-100-insoluble material was introduced into the reconstituted liposomes, the vesicles became permeable to sucrose, raffinose (Mr, 504), and stachyose (Mr, 666) and fully retained dextrans of Mr greater than 1,500. Inulin (average Mr, 1,400) was tested for its efflux from the reconstituted outer membrane vesicles; 62% of the added inulin was trapped. The molecular weight exclusion limit for the outer membrane of H. influenzae type b was therefore estimated at approximately 1,400. A protein responsible for the transmembrane diffusion of solutes was purified from H. influenzae type b by extraction of whole cells with cetyl trimethyl ammonium bromide. When this extract was passed over DEAE-Sepharose, three protein-containing peaks (I, II, and III) were eluted. Peaks I and II contained mixtures of proteins as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; when tested for their pore-forming properties, these proteins were unable to render liposomes of lipopolysaccharide and phospholipid permeable to sucrose. Peak III contained only one molecular species of protein of molecular weight 40,000; this protein acted as a porin in reconstituted vesicles. The molecular weight exclusion limit for 40,000-molecular-weight protein matched the estimate of approximately 1,400 which was determined for outer membranes. A series of homologous saccharides of increasing degree of polymerization was prepared from agarose by hydrolysis with beta-agarase and fractionation on gel filtration chromatography. These oligosaccharides of Mr, 936, 1,242, 1,548, and 1,854 were assayed for retention by the complete vesicles containing 40-kilodalton protein and lipopolysaccharide and phospholipids. All of these oligosaccharides were lost by efflux through the porin. Since the molecular conformation of the largest oligosaccharide is an elongated semirigid helix, it is suggested that the pore formed by the 40-kilodalton protein does not act as a barrier to the diffusion of this compound.
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PMID:Transmembrane permeability channels across the outer membrane of Haemophilus influenzae type b. 298 94

The mechanism(s) by which the lipopolysaccharide (LPS) of Haemophilus influenzae type b may contribute to the virulence of this organism is unclear. Purified LPS of Haemophilus influenzae type b or phosphate buffered saline was administered intranasally to infant rats prior to the intranasal instillation of approximately 2-20 x 10(6) cfu of Hib two or three times per day for three consecutive days. The preadministration of 2.0 micrograms Hib LPS resulted in a significantly greater incidence of bacteremia (P = 0.0006) than PBS 30 min after the completion of the intranasal inoculation. Four days following completion of intranasal Hib inoculation the incidence of bacteremia was greater (P = 0.017) in the animals pretreated with LPS at 2.0 micrograms compared to the PBS pretreated animals. Preadministration of 0.2 micrograms LPS had no effect on the incidence of bacteremia or meningitis. There were no differences in the histology of the nasal cavities or turbinates of infant rats inoculated intranasally only with LPS or PBS. There were no differences in the frequency or density of bacteremia following intranasal administration of LPS from either Hib or E. coli. Although the mechanism is unknown, our findings suggest that the LPS of Hib may contribute to the ability of H. influenzae type b to invade the nasal mucosa in this infant rat model.
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PMID:Contribution of Haemophilus influenzae type b lipopolysaccharide to pathogenesis of infection. 307 63

In the course of using the infant rat model to determine the ability of various rabbit antisera to protect against challenge by Haemophilus influenzae type b we made two unexpected observations. In these experiments 4-day-old rats were inoculated s.c. on the dorsum with either rabbit serum or physiological buffers (sham serum) and then were challenged the next day with H. influenzae type b injected i.p. Bacteremia, as a marker for disease, was measured 24 h later on day 6. We observed the following. (i) Pre-immune, i.e., normal rabbit serum, containing minimal levels of antibodies to outer membrane proteins and depleted of antibodies to capsule and lipopolysaccharide, nevertheless significantly (P less than 0.01) protected the rats from challenge with H. influenzae type b when compared to a sham inoculation of buffer; (ii) In the absence of a serum inoculation on day 4 (a buffer was used as a sham serum inoculation), the levels of bacteremia obtained after inoculation with bacteria on day 5 depended upon the composition of the buffer in which the H. influenzae inoculum was suspended. Use of phosphate buffered saline (PBS) resulted in higher levels of bacteremia than PBS containing 0.5% bovine serum albumin (PBS-BSA) (P less than 0.001), i.e. the BSA apparently acted to protect the rats from H. influenzae infection. In fact the use of PBS-BSA as an inoculum buffer masked the protective effect noted above of the absorbed normal rabbit serum.
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PMID:Unexpected effects of absorbed normal rabbit serum and bovine serum albumin on survival of Haemophilus influenzae type b in the infant rat. 326 78

Ribosomal preparations from Klebsiella pneumoniae, Haemophilus influenzae, Streptococcus pyogenes, and Streptococcus pneumoniae were investigated with respect to their activating capacity towards murine lymphoid cells. The proliferation of BALB/c spleen cells was induced in a dose-dependent fashion (from 1 to 100 micrograms/ml) by ribosomes of K. pneumoniae, H. influenzae, and S. pyogenes with a peak activity at 48 or 72 hr of culture. The majority of the blast cells induced by these ribosomal preparations were positive for surface-immunoglobulin (S-Ig) and negative for Thy 1.2. Furthermore, K. pneumoniae, H. influenzae, and S. pyogenes ribosomes induced the synthesis of IgM and some IgA. Cell proliferation and induction of IgM production were also demonstrated with the 3 ribosomal preparations using spleen cells from athymic nude (nu+/nu+) mice, Lyb-5-defective CBA/N spleen cells, B cell-enriched and T cell-depleted BALB/c spleen cell suspensions, as well as spleen cells from the Ips gene-deficient C3H/HeJ strain. Cell culture supernatants contained specific anti-ribosome IgM antibodies. Antibodies of other specificities (anti-sheep erythrocytes) were also demonstrated in supernatants from K. pneumoniae-stimulated cultures. Evidence against a possible role of contamination of K. pneumoniae and H. influenzae ribosomes by lipopolysaccharide- or lipid A-associated proteins in this effect is discussed. Ribosomes from S. pneumoniae did not induce 3H-thymidine incorporation nor Ig production. None of the 4 ribosomal preparations was found to stimulate T cell blastogenesis or to induce interleukin-2 production by naive BALB/c spleen cells. Finally, ribosomes from H. influenzae, S. pyogenes, S. pneumoniae but not those of K. pneumoniae stimulated interleukin-1 production by adherent spleen cells, from BALB/c mice.
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PMID:Induction of murine B cell proliferation and immunoglobulin synthesis by some bacterial ribosomes. 326 81

The chinchilla experimental model of otitis media was used to examine the importance of serum antibodies in protection against disease caused by nontypable Haemophilus influenzae. An immune serum pool was prepared by immunizing chinchillas with killed bacterial cells of nontypable H. influenzae 3245. Pooled preimmune or immune serum from these immunized animals was administered intravenously to a group of nonimmune chinchillas 1 day before intrabullar challenge with strain 3245. Of 5 animals receiving preimmune serum, 5 developed otitis media compared with 0 of 10 animals receiving immune serum (P = 0.008). The immune serum pool contained antibodies directed against both surface-exposed outer membrane proteins and lipopolysaccharide (LPS). The 39-kilodalton major outer membrane protein was the immunodominant surface protein. Anti-LPS antibodies were removed from the immune serum pool by affinity chromatography, and affinity-purified anti-LPS antibodies were recovered. Immune serum, immune serum absorbed of LPS antibodies, or affinity-purified LPS antibodies were then administered to another group of experimental animals 1 day before bacterial challenge. Of four animals that received the affinity-purified LPS antibodies, four developed otitis compared with zero of four animals that received the immune serum or zero of four animals that received the LPS-absorbed immune serum (P = 0.028). These studies indicate that passive immunization with immune serum is protective in experimental nontypable H. influenzae otitis media and that bacterial outer membrane proteins may be the principal targets of protective antibody.
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PMID:Protection by serum antibodies in experimental nontypable Haemophilus influenzae otitis media. 348 58

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of lipopolysaccharide (LPS) was performed to assess the usefulness of this technique for the epidemiologic analysis of Haemophilus influenzae type b isolates. LPS samples were prepared from isolates which had been passaged either in vitro or in infant rats. Preparations from paired isolates from a number of epidemiologically related clinical situations also were examined. The gel patterns of LPS prepared on different occasions from an individual isolate were stable. However, the LPS gel patterns changed in 5 of 14 (36%) of the passaged isolates, and differences in gel patterns also were observed among epidemiologically related isolates. The variability in LPS electrophoretic patterns of individual isolates indicated that this technique is not useful for the epidemiologic analysis of H. influenzae type b disease.
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PMID:Lipopolysaccharide gel profiles of Haemophilus influenzae type b are not stable epidemiologic markers. 348 5

Protein a (46,000 molecular weight [46K]) was purified from outer membranes of Haemophilus influenzae type b by a relatively simple procedure. Spontaneously shed outer membranes from a 24-h, 12-liter culture of an unencapsulated variant of strain Eag were combined with outer membranes released from the cells by Tris buffer and extracted with the nonionic detergent octylpolyoxyethylene. The extract was then subjected to open column chromatography on Sephacryl S-200 and Trisacryl-carboxymethyl to yield 7.5 mg of protein a from 180 mg of outer membrane protein. Approximately 99% of the protein in this preparation was protein a; in addition, the preparation contained 1.25% (wt/wt) lipopolysaccharide and had a residual detergent/protein ratio of 1.6:1 (wt/wt). Antibodies to the preparation were induced in rabbits by using alum as an adjuvant. As determined by immunoblotting, the great preponderance of antibodies induced were specific for protein a. However, very low levels of antibodies to several other outer membrane components, which were not apparent on gels of the pure preparation of protein a, were also induced. Preimmune and postimmune sera, after depletion of antibodies to capsular polysaccharide and lipopolysaccharide, were tested for biological activity against H. influenzae type b. Compared with preimmune serum, postimmune serum was bactericidal in vitro against strain Eag (the only strain tested) and offered significant protection (P less than 0.01) to infant rats against infection by all four strains tested, two of which had a protein a that was larger (47K) than the 46K protein a in the preparation. These results indicate that protein a should be considered as a vaccine to prevent H. influenzae type b disease.
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PMID:Protection of infant rats from Haemophilus influenzae type b infection by antiserum to purified outer membrane protein a. 349 97

Strains of Haemophilus influenzae isolated in The Netherlands between 1975 and 1984 from patients with meningitis were analysed in order to determine whether older patients are infected with particular types or subtypes of the organism. Of 1154 patients with H. influenzae meningitis 73 (6.3%) were more than 6 years of age. Thirty-one strains (42%) were of serotype b, one strain was of serotyped, one strain was of serotype f and 40 strains (55%) were non-typable. Twenty-eight type b strains were available for subtyping by analysis of the major outer-membrane proteins by sodium dodecylsulphate (SDS)-polyacrylamide gel electrophoresis (PAGE), by serotyping of their lipopolysaccharides and by biotyping. Twenty-one strains were outer-membrane protein subtype 1,24-lipopolysaccharide serotype 1 and 24 biotype I. Seventeen strains (61%) combined these characteristics. This percentage did not differ significantly from the percentage found for strains isolated from patients of all age groups (80%). The 32 non-typable H. influenzae strains analysed had different outer-membrane protein patterns as seen by SDS-PAGE. Five biotypes were found, among which biotype II was predominant (21/32). The results indicated that (i) patients more than 6 years of age were infected by subtypes of H. influenzae b strains which were not significantly different from the strains isolated from younger patients, (ii) non-typable strains of H. influenzae were much more common (55%) in the older age group than in the younger (1.2%) and (iii) that these non-typable strains were not of a particular subtype.
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PMID:Types and subtypes of 73 strains of Haemophilus influenzae isolated from patients more than 6 years of age with meningitis in The Netherlands. 349 69

The techniques of capsular serotype, biotype determination and sodium dodecylsulphate polyacrylamide gel electrophoresis of sarcosinate-insoluble outer membrane protein (OMP) and of proteinase K lipopolysaccharide (LPS) preparations were applied to 41 genital and neonatal Haemophilus influenzae isolates. Twelve percent were capsulated (4b, 1a). Distribution of strains between biotypes was similar to that of isolates of other non-systemic pathogenic origin; only one isolate was biotype IV. The OMP profiles showed great variability with 4 group of proteins: a 16-Kd major peptide which was observed in all strains; 27-30-Kd major OMP including one constant 30-Kd peptide present in all strains except one; 32- to 50-Kd major OMP; and 49- to 54-Kd minor OMP. The rough LPS profiles also revealed heterogeneity. In view of the variability observed among H. influenzae strains, it is not possible to establish a relationship between pathogenicity and a macromolecular marker.
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PMID:[Heterogeneity of strains of Haemophilus influenzae of genital and neonatal origin: analysis of capsular serotypes, biotypes and electrophoretic profiles of external membrane proteins and LPS]. 350 Jul 32


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