Isolation and partial characterization of a lipopolysaccharide from phase II <i>Coxiella burnetii</i>

Baca, O G; Martinez, Irene; Aragon, A S; Klassen, D A

doi:10.1139/m80-141

Cited by 15 publications

(11 citation statements)

References 26 publications

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“…The C. burnetii LPSs described here were found to contain a KDO-like, thiobarbituric acid reactive substance and heptose in amounts comparable to previously reported phase I and II LPS composition (2). Also as previously reported for phase I and II LPSs (2,16), all three C. burnetii LPSs gelled LALs at subnanogram levels. The C. burnetii LPSs have been considered endotoxic (3,34).…”

Section: Discussionsupporting

confidence: 87%

Lipopolysaccharide variation in Coxiella burnetti: intrastrain heterogeneity in structure and antigenicity

Hackstadt¹,

Peacock²,

Hitchcock³

et al. 1985

Infect Immun

169

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We isolated lipopolysaccharides (LPSs) from phase variants of Coxiella burnetii Nine Mile and compared the isolated LPS and C. burnetii cells by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The LPSs were found to be the predominant component which varied structurally and antigenically between virulent phase I and avirulent phase II. A comparison of techniques historically used to extract the phase I antigenic component revealed that the aqueous phase of phenol-water, trichloroacetic acid, and dimethyl sulfoxide extractions of phase I C. burnetii cells all contained phase I LPS, although the efficiency and specificity of extraction varied. Our studies provide additional evidence that phase variation in C. burnetii is analogous to the smooth-to-rough LPS variation of gram-negative enteric bacteria, with phase I LPS being equivalent to smooth LPS and phase II being equivalent to rough LPS. In addition, we identified a variant with a third LPS chemotype with appears to have a structural complexity intermediate to phase I and II LPSs. All three C. burnetii LPSs contain a 2-keto-3-deoxyoctulosonic acid-like substance, heptose, and gel Limulus amoebocyte lysates in subnanogram amounts. The C. burnetii LPSs were nontoxic to chicken embryos at doses of over 80 ,ug per embryo, in contrast to Salmonella typhimurium smooth-and rough-type LPSs, which were toxic in nanogram amounts. Coxiella burnetii, the etiologic agent of Q fever, is an obligately intracellular bacterium that multiplies within the

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Section: Discussionsupporting

confidence: 87%

Lipopolysaccharide variation in Coxiella burnetti: intrastrain heterogeneity in structure and antigenicity

Hackstadt¹,

Peacock²,

Hitchcock³

et al. 1985

Infect Immun

169

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“…Recent studies (3, 29) of the structures of LPS I and phase II LPS (LPS II) of C. burnetii produced results which were somewhat contradictory. The results of Baca et al (3) suggest that LPS I and LPS II have similar sugar compositions and serological activities. However, the more recent study by Schramek and Meyer (31) confirms previous indications (29) of LPS sugar components but reveals distinctly different sugar compositions.…”

mentioning

confidence: 94%

Chemical and immunological characterization of lipopolysaccharides from phase I and phase II Coxiella burnetii

Amano¹,

Williams²

1984

J Bacteriol

100

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Lipopolysaccharides (LPSs) isolated from phase I and phase II Coxiella burnetii (LPS I and LPS II, respectively) were analyzed for chemical compositions, molecular heterogeneity by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, and immunological properties. The yields of crude phenol-water extracts from phase I cells were roughly three to six times higher than those from phase II cells. Purification of LPSs by ultracentrifugation gave similar yields for both LPS I and LPS II. Purified LPS I and LPS II contained roughly 0.8 and 0.6% protein, respectively. The fatty acid constituents of the LPSs were different in composition and content, with branched-chain fatty acids representing about 15% of the total. I8-Hydroxymyristic acid was not detected in either LPS I or LPS II. A thiobarbituric acid-periodate-positive compound was evident in the LPSs; however, this component was not identified as 3-deoxy-D-mannooctulosonic acid by gas and paper chromatographies. LPS II contained D-mannose, D-glucose, D-glyceromannoheptose, glucosamine, ethanolamine, 3deoxy-D-mannooctulosonic acid-like material, phosphate, and fatty acids. LPS I contained the unique disaccharide galactosaminuronyl glucosamine and nine unidentified components in addition to the components of LPS II. The hydrophobic, putative lipid A fraction of LPS I and LPS II contained the above constituents, but the hydrophilic fraction was devoid of ethanolamine. The LPS I disaccharide galactosaminuronyl glucosamine was found in both fractions of the acetic acid hydrolysates. Analysis of LPSs by sodium dodecyl sulfatepolyacrylamide gel electrophoresis followed by silver staining indicated that LPS II was composed of only one band, whereas LPS I consisted of six or more bands with irregular spacing. Ouchterlony immunodiffusion tests demonstrated that LPS I reacted with phase I but not with phase II whole-cell hyperimmune antibody, and LPS II reacted neither with phase I nor phase II hyperimmune antibody. From these results, it was concluded that the chemical structures of LPSs from C. burnetii were different from those of the LPSs of gram-negative bacteria; however, the LPS structural variation in C. burnetii may be similar to the smooth-to-rough mutational variation of saccharide chain length in gram-negative bacteria.

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“…Differences in surface properties between the virulent and avirulent phases (Fiset & Ormsbee, 1968;Krauss et al, 1977) and in the carbohydrate composition of phase 1 and phase 2 antigens (Schramek & Brezina, 1979;Baca et al, 1980) have led investigators to compare this phase variation with the smooth-to-rough transition of various enterobacteria. Demonstration of quantitative and qualitative differences in the sugar compositions of lipopolysaccharides (LPS) isolated from the two phases supports this comparison (Schramek & Mayer, 1982).…”

Section: Introductionmentioning

confidence: 99%

Stability of Plasmid Sequences in an Acute Q-Fever Strain of Coxiella burnetii

Samuel

Frazier²,

Mallavia³

1988

Microbiology

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1795The rickettsia1 pathogen Coxiella burnetii undergoes a variation in which virulent isolates (phase 1) become avirulent (phase 2) after repeated passage in a non-immunologically competent host. Biochemically, this variation is associated with a lipopolysaccharide modification and possibly other factors. Genetically, the regions of DNA responsible for phase variation have not been identified. We have sought to determine whether the plasmid identified in acute disease isolates, QpH 1, which represents approximately 5 % of the coding capacity of this organism is involved in phase variation. Plasmids from phase 1 and phase 2 variants (designated QpHl and QpH2, respectively) were compared by restriction endonuclease digestion and Southern blot hybridization to determine whether sequence changes in the phase 2 plasmid might account for changes in the virulence of phase 2 organisms compared with that of phase 1 cells. Using over 20 different restriction enzymes, no changes in DNA restriction fragment patterns were detected regardless of whether the phase change occurred during egg or tissue culture passage. The plasmid-specific mRNAs produced from metabolically active, purified cells were identical for each phase type. Using QpHl or QpH2 DNA as a template, the mRNA produced by an E. coli extract was also identical. Finally, the proteins encoded by either plasmid in an in vitro transcription/translation reaction were identical. These data indicate that within the limits of our analysis, the plasmid DNA from C. burnetii phase variants is structurally and functionally the same and is therefore unlikely to be involved in phase variation.

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Isolation and partial characterization of a lipopolysaccharide from phase II Coxiella burnetii

Cited by 15 publications

References 26 publications

Lipopolysaccharide variation in Coxiella burnetti: intrastrain heterogeneity in structure and antigenicity

Lipopolysaccharide variation in Coxiella burnetti: intrastrain heterogeneity in structure and antigenicity

Chemical and immunological characterization of lipopolysaccharides from phase I and phase II Coxiella burnetii

Stability of Plasmid Sequences in an Acute Q-Fever Strain of Coxiella burnetii

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