Antibodies to Core Lipopolysaccharide Determinants: Absence of Cross-reactivity with Heterologous Lipopolysaccharides

Heumann, Didier; Baumgartner, J. D.; Jacot-Guillarmod, H.; Glauser, M. P.

doi:10.1093/infdis/163.4.762

Cited by 48 publications

(16 citation statements)

References 62 publications

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“…IgGs were isolated from preimmune (control IgG) and immune (anti-LBP IgG) plasma by protein A chromatography. Since we used LPS isolated from Escherichia coli 0111 (Sigma) in our experiments, we ruled out the presence of anti-LPS 0111 antibodies in this plasma by ELISA (11) (data not shown). F(ab')2 fragments were obtained by trypsin digestion (Pierce).…”

Section: Methodsmentioning

confidence: 99%

Mode of action of anti-lipopolysaccharide-binding protein antibodies for prevention of endotoxemic shock in mice.

Gallay¹,

Heumann²,

Roy³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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Lipopolysaccharide (LPS)-binding protein (LBP) has been shown to regulate the response of monocytes to LPS in vitro. In a previous study, polyclonal anti-LBP IgGs were found to protect D-galactosamine-sensitized mice against a lethal endotoxemic shock induced by a low challenge of LPS or lipid A when administered simultaneously with endotoxin.In the present study, we investigated the mode of action ofthese anti-LBP IgGs. In vitro, we demonstrated that they interfere with LPS binding to Lipopolysaccharide (LPS)-binding protein (LBP) isolated from the blood of humans, rabbits, and mice has been shown to bind with a high affinity (Kd = 1 nM) to lipid A, the common moiety of LPS (1-3). LBP-LPS complexes rapidly bind to receptor CD14 expressed on the surface of monocytic cells and on activated neutrophils, triggering cell activation (2-6). In the presence of LBP, mediators including cytokines and nitrites, are released at concentrations of LPS far below those required for cell stimulation exposed to LPS in the absence of LBP (2-7). Antibody-mediated blockade of LBP or CD14 prevents the in vitro LPS-induced cytokine release by cells harboring CD14 on their surface (4-8). Therefore, it was postulated that LBP and CD14 represent a pair of key molecules in initiation of host defense mechanisms during infections caused by Gram-negative organisms (9, 10).Recently, we reported that treatment with an IgG preparation raised against purified murine LBP protected D-galactosamine-sensitized mice against a lethal endotoxemic shock induced by a low challenge of LPS or of lipid A (8). This protection was observed when the anti-LBP IgG was given simultaneously with LPS challenge, and it was associated with a strong decrease of circulating tumor necrosis factor (TNF). We have shown in parallel that type-specific anti-LPS antibodies afforded similar protection and decreased TNF to the same extent as the anti-LBP IgG (8).In the present study, we first characterized in vitro how anti-LBP IgG affected the binding of LPS to monocytes and polymorphonuclear cells (PMNs). We next investigated how and by which mechanisms antibodies present in the polyclonal anti-LBP IgG preparation could be involved in protecting mice from endotoxemic shock. MATERIALS AND METHODSAntibodies. Anti-LBP antibodies were raised in rabbits, using purified murine LBP as immunogen as described (3,8). IgGs were isolated from preimmune (control IgG) and immune (anti-LBP IgG) plasma by protein A chromatography. Since we used LPS isolated from Escherichia coli 0111 (Sigma) in our experiments, we ruled out the presence of anti-LPS 0111 antibodies in this plasma by ELISA (11) (data not shown). F(ab')2 fragments were obtained by trypsin digestion (Pierce). Anti-O111 LPS IgGs were isolated by protein A chromatography from serum of a rabbit immunized with E. coli 0111 bacteria (12).Plasma and Cells. Heparin-treated blood samples obtained from human volunteers were used to prepare PMNs and peripheral blood mononuclear cells (PBMCs) by centrifugation over Ficoll/Hypaqu...

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Section: Methodsmentioning

confidence: 99%

Mode of action of anti-lipopolysaccharide-binding protein antibodies for prevention of endotoxemic shock in mice.

Gallay¹,

Heumann²,

Roy³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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“…Because the observed antibody binding to the various lipopolysaccharide preparations could be nonspecific, several additional experiments were performed. Blocking hydrophobic domains on lipopolysaccharide by complexing to other molecules has been used to suppress or eliminate nonspecific binding (1,4,23,48). Purified P. gingivalis and B. forsythus lipopolysaccharide were complexed to bovine serum albumin, polymyxin-B and apolipoprotein Al as described in the Material and methods section, and compared by ELISA to naked, uncomplexed lipopolysaccharide.…”

Section: Resultsmentioning

confidence: 99%

“…If a significant proportion of the observed binding were nonspecific, binding would have occurred from the preimmune as well as the immune rabbit serum and that was not the case. Nonspecific binding can be suppressed by complexing lipopolysaccharide to bovine serum albumin, apolipoprotein Al (one of the proteins that binds highdensity lipoproteins to lipopolysaccharide), and polymyxin-B (5,23,48). Interpretation of the results of such experiments, however, is made difficult by the fact that some of these molecules such as polymyxin-B also bind to antigenic epitopes in lipid A (3,13).…”

Section: Discussionmentioning

confidence: 99%

Shared antigens of Porphymmonas gingivalis and Bacteroides forsythus

Vasel

Sims

Bainbridge

et al. 1996

Oral Microbiology and Immunology

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Periodontitis in humans is caused by a group of predominantly gram-negative, anaerobic bacteria among which Porphyromonas gingivalis and Bacteroides forsythus are prominent. A similar group is present and presumably plays a similar role in experimental periodontitis in the primate Macaca fascicularis. Nevertheless, immunization using a vaccine containing only killed P. gingivalis suppresses the progress of experimental periodontitis in M. fascicularis. We investigated the hypothesis that gram-negative periodontopathic bacterial may share antigens, and immunization with one species may induce antibodies reactive with other gram-negative species. Using enzyme-linked immunosorbent assay (ELISA), Western and dot immunoblots with nonabsorbed and absorbed and immune and preimmune sera we show that monkeys immunized with P. gingivalis produce antibodies reactive not only with antigens of P. gingivalis but also with those of B. forsythus. Similarly, rabbits immunized with P. gingivalis or with B. forsythus produce antibodies that react with antigens of both bacteria. Cross-reactive antibodies bind to epitopes in lipid A and possibly in core carbohydrate of lipopolysaccharide. Using complexes of lipopolysaccharide with polymyxin B, bovine serum albumin and apolipoprotein A1 specificity of binding was documented. Using sera from monkeys immunized with P. gingivalis, cross-reactivity with Actinobacillus actinomycetemcomitans could not be demonstrated by ELI-SA, although binding to lipopolysaccharide but not to lipid A was demonstrated by Western and dot immunoblots. Antibodies to shared lipopolysaccharide epitopes of periodontopathic bacteria may account, at least in part, for the immune protection observed in immunized monkeys, and shared epitopes may have potential as a vaccine for periodontitis in humans.

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“…D. Baumgartner, unpublished data). By solubilization of the core glycolipid in a physiological manner to circumvent nonspecific binding [19], the 15 antisera used in one study were later shown to contain IgG and IgM antibody to 15 at a titer only threefold higher than that in control (preimmune) serum; furthermore, these antisera contained no more antibodies to lipid A than did the control serum [20]. Moreover, it now appears that antibodies to 15 are highly specific for E. coli15 and that they do not cross-react with endotoxin from other bacteria.…”

Section: Antibodies To Endotoxinmentioning

confidence: 99%

Pathogenesis and Potential Strategies for Prevention and Treatment of Septic Shock: An Update

Glauser

Heumann

Baumgartner

et al. 1994

Clinical Infectious Diseases

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104

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Antibodies to Core Lipopolysaccharide Determinants: Absence of Cross-reactivity with Heterologous Lipopolysaccharides

Cited by 48 publications

References 62 publications

Mode of action of anti-lipopolysaccharide-binding protein antibodies for prevention of endotoxemic shock in mice.

Mode of action of anti-lipopolysaccharide-binding protein antibodies for prevention of endotoxemic shock in mice.

Shared antigens of Porphymmonas gingivalis and Bacteroides forsythus

Pathogenesis and Potential Strategies for Prevention and Treatment of Septic Shock: An Update

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