Biological activities of fragments derived from Bordetella pertussis endotoxin: isolation of a nontoxic, Shwartzman-negative lipid A possessing high adjuvant properties

Aymé, Gabriel; Caroff, Martine; Chaby, Richard; Haeffner‐Cavaillon, Nicole; Dur, Annick Le; Moreau, Monique; Muset, M; Mynard, M. C.; Roumiantzeff, M; Schulz, Dominique; Szabó, Ladislas

doi:10.1128/iai.27.3.739-745.1980

Cited by 46 publications

(14 citation statements)

References 25 publications

(19 reference statements)

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“…The efficiency of two different procedures aiming at the removal of SDS from LPS contaminated with the detergent was evaluated using radioactive SDS. Pertussis endotoxin (20 mg) was kept for 5 min at 1000C in 1% [3S]SDS (2 ml, 3.08 x 1.07 cpm). Molar sodium phosphate (pH 6.4, 2.25 ml) was added to a portion (250 pl) and the mixture was then dialyzed, for 24 h, against distilled water (15 ml) which was replaced 6 times during the first 6 h. One hundred percent of the endotoxin but only 1.5% of the radioactivity was recovered in the nondiffusible material at the end of the dialysis procedure.…”

Section: Resultsmentioning

confidence: 99%

Isolation of two protein-free and chemically different lipopolysaccharides from Bordetella pertussis phenol-extracted endotoxin

Dur¹,

Chaby²,

Szabó³

1980

J Bacteriol

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Endotoxin prepared from several Bordetella pertussis strains in both immunological phases I and IV gave two lipopolysaccharide peaks (LPS-I and LPS-II) when analyzed on hydroxylapatite columns in a phosphate buffer containing 0.1% sodium dodecyl sulfate; these lipopolysaccharides, present in the ratio of 2:3, are true endotoxins by both chemical and biological criteria. Endotoxins isolated from Escherichia coli, Salmonella typhimurium, and Shigella flexneri gave single lipopolysaccharide peaks when analyzed by the same procedure. Upon hydrolysis with acetic acid (pH 3.4) at 100 degrees C for 1 h, LPS-I released a polysaccharide (PS-I); the linkage broken was that of the glycosidic bond of a non-phosphorylated 3-deoxy-oct-2-ulosonic acid. Treatment with 0.25 M mineral acid at 100 degrees C for 30 min was required to free the polysaccharide moiety (PS-II) of LPS-II, the linkage broken being the glycosidic bond of a phosphorylated 3-deoxy-oct-2-ulosonic acid. Chemical and physical differences of the polysaccharide moieties PS-I and PS-II present in LPS-I and LPS-II have been described previously (25). By using the technique of 125I labeling, it was shown that the totality of labeled proteins present in the endotoxin extracted from Bordetella pertussis by the phenol-water procedure could be separated from the lipopolysaccharide by column chromatography on hydroxylapatite; it follows that these proteins are not linked by covalent bonds to the lipopolysaccharide.

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Section: Resultsmentioning

confidence: 99%

Isolation of two protein-free and chemically different lipopolysaccharides from Bordetella pertussis phenol-extracted endotoxin

Dur¹,

Chaby²,

Szabó³

1980

J Bacteriol

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“…Cell growth. Culture conditions of Bordetella pertussis cells in the liquid medium of Cohen & Wheeler (1946), extraction of the endotoxin by the phenol-water procedure (Westphal et al, 1952), and its purification by sedimentation have been described previously (Ayme et al, 1980). Fntfy acid extructzon.…”

Section: Methodsmentioning

confidence: 99%

“…In view of the foregoing, and of our current interest in the chemical structure and immunological activities (Ayme et al, 1980;Haeffner-Cavaillon et al, 1982 of the Bordetellapertussis endotoxin, a quantitative study was undertaken of the fatty acids present in this endotoxin, and the conditions of their release; the results are reported in this paper.…”

Section: Introductionmentioning

confidence: 99%

The Fatty Acid Content of the Bordetella pertussis Endotoxin

Starkloff¹,

Szabó

1986

Microbiology

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The fatty acid content of Bordetella pertussis endotoxin has been estimated by several methods. Expressed as 3-hydroxytetradecanoic acid, it was 0.74 mumol (mg lyophilized material)-1, 0.38 mumol being ester-bound, and 0.32 mumol in amide linkage. Reported molar ratios of ester-bound to amide-bound fatty acids in endotoxins of various bacterial species range from 2.4 to 2 in B. pertussis, to 5 to 2 in Salmonella minnesota; according to these figures large differences must exist in the degree of substitution, and the substitution pattern of the glucosaminyl-beta-1,6-glucosamine unit present in the hydrophobic region of endotoxins. When fatty acids, released by acid and alkaline hydrolyses of the B. pertussis endotoxin, were extracted into chloroform, unidentified chromogenic substances appearing in the extract interfered with their colorimetric estimation; no interference was observed when hexane was used instead of chloroform.

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“…(B) N. meningitidis. Lanes 1,2, and 3 correspond, respectively, to LPS, crude lipid A liberated from LPS by hydrolysis in acetate buffer (pH 4.5), and purified lipid A (acetate buffer [pH 4.5]). (C) E. coli.…”

Section: Downloaded Frommentioning

confidence: 99%

Inability of pyrogenic, purified Bordetella pertussis lipid A to induce interleukin-1 release by human monocytes

Caroff¹,

Cavaillon²,

Fitting³

et al. 1986

Infect Immun

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Free lipid A of BordeteUa pertussis, Neisseria meningitidis, and Escherichia coli lipopolysaccharide (LPS) was prepared by hydrolysis in acetate buffer (pH 4.5); in addition, lipid A from B. pertussis and E. coli was prepared by hydrolysis in mineral acid (HCI). The precipitates obtained were purified by extraction methods in toluene-methanol and are referred to as crude lipid A. Purified lipid A from N. meningitidis and B. pertussis was obtained by extraction in a mixture of chloroform-methanol-water-triethylamine. The different preparations were tested for their pyrogenicity (endogenous pyrogen; EP) and their capacity to trigger the release of interleukin-l (IL-1; previously known as lymphocyte-activating factor; LAF) by human monocytes. Crude lipid A from E. coli and N. meningitidis were both IL-1 inducers. Crude B. pertussis lipid A (acetate buffer; pH 4.5), which contains a 3-1-6-linked D-glucosamine disaccharide, two phosphoryl groups, and five fatty acids, was pyrogenic and an IL-1 inducer (EP+/LAF+); but crude B. pertussis lipid A (0.25 N HCI), which lacked the glycosidic phosphoryl group, was 1,000-fold less pyrogenic than the diphosphorylated lipid A, yet it retained its ILl inducing capacity (EP-/LAFt). Purified N. meningitidis lipid A was not an inducer of IL-1 release and purified B. pertussis lipid A exhibited identical pyrogenicity as the parent LPS but was devoid of any ILl release inducing capacity (EP /LAF-). These results demonstrate that for some endotoxins, purified lipid A is unable to induce IL-1 release by human monocytes; however, it is pyrogenic, supporting the hypothesis that IL-1 and EP are induced by different determinants on the LPS molecule.

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Biological activities of fragments derived from Bordetella pertussis endotoxin: isolation of a nontoxic, Shwartzman-negative lipid A possessing high adjuvant properties

Cited by 46 publications

References 25 publications

Isolation of two protein-free and chemically different lipopolysaccharides from Bordetella pertussis phenol-extracted endotoxin

Isolation of two protein-free and chemically different lipopolysaccharides from Bordetella pertussis phenol-extracted endotoxin

The Fatty Acid Content of the Bordetella pertussis Endotoxin

Inability of pyrogenic, purified Bordetella pertussis lipid A to induce interleukin-1 release by human monocytes

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