Conserved Epitopes in Bacterial Lipopolysaccharides and Cross‐Reactive Antibodies in the Treatment of Endotoxicosis and Gram‐Negative Septic Shock

Müller‐Loennies, Sven; Padova, Franco Di; Brade, Lore; Heumann, Didier; Rietschel, Ernst Theodor

doi:10.1002/9780470695951.ch9

Cited by 5 publications

(7 citation statements)

References 159 publications

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“…were assigned to a b-GlcpN (164 Hz, residue B) and b-Galp (164 Hz, residue M), respectively. Their b-configuration was confirmed by their 3 J H,H-coupling constants ( 8 Hz) and their intraresidual NOE connectivities between H-1, H-3 and H-5. Three signals of anomeric protons showed 3 J H,H-coupling constant values of less than 1 Hz and thus the H-2 in these residues was in equatorial position as in manno-configurated sugars (residues E, F, H).…”

Section: R E S U L T Smentioning

confidence: 89%

“…It was assumed that these antibodies would be both cross-reactive and crossprotective against different Gram-negative pathogens. Whereas a cross-protective effect was described for a polyclonal antiserum as early as in 1966 [4], all subsequently isolated monoclonal antibodies failed to show cross-reactivity in vitro and cross-protectivity in vivo [3], except one reported by DiPadova et al . This mAb recognized LPS from all tested clinical isolates of E. coli, Salmonella, and Shigella in Western-blots and showed cross-protective effects in vivo against endotoxic activities of LPS [5].…”

mentioning

confidence: 99%

“…This prompted many investigators to attempt the isolation of antibodies directed against the conserved regions of LPS, i.e. lipid A and core‐region (reviewed in [3]). It was assumed that these antibodies would be both cross‐reactive and cross‐protective against different Gram‐negative pathogens.…”

mentioning

confidence: 99%

“…It was assumed that these antibodies would be both cross‐reactive and cross‐protective against different Gram‐negative pathogens. Whereas a cross‐protective effect was described for a polyclonal antiserum as early as in 1966 [4], all subsequently isolated monoclonal antibodies failed to show cross‐reactivity in vitro and cross‐protectivity in vivo [3], except one reported by DiPadova et al . (mAb WN1 222‐5).…”

mentioning

confidence: 99%

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Structural analysis of deacylated lipopolysaccharide of Escherichia coli strains 2513 (R4 core‐type) and F653 (R3 core‐type)

Müller‐Loennies

Lindner

Brade

2002

European Journal of Biochemistry

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Lipopolysaccharide (LPS) of Escherichia coli strain 2513 (R4 core-type) yielded after alkaline deacylation one major oligosaccharide by high-performance anion-exchange chromatography (HPAEC) which had a molecular mass of 2486.59 Da as determined by electrospray ionization mass spectrometry. This was in accordance with the calculated molecular mass of a tetraphosphorylated dodecasaccharide of the composition shown below. NMR-analyses identified the chemical structure as We have also isolated the complete core-oligosaccharides of E. coli F653 LPS for which only preliminary data were available and investigated the deacylated LPS by NMR and MS. The proposed structure determined previously by methylation analysis was confirmed and is shown below.In addition we have quantified the side-chain heptose substitution of the inner core with GlcpN ( 30%) and confirmed that this sugar is only present when the phosphate at the second L,D-Hepp residue is absent.

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Section: R E S U L T Smentioning

confidence: 89%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Structural analysis of deacylated lipopolysaccharide of Escherichia coli strains 2513 (R4 core‐type) and F653 (R3 core‐type)

Müller‐Loennies

Lindner

Brade

2002

European Journal of Biochemistry

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“…20 This strain has been claimed, since the late 1960s, to induce Abs in mice, rabbits and humans that have a broad serological cross-reactivity and a broad cross-neutralizing activity against LPS of Gram-negative bacteria in general. 21 Even various mAbs have been obtained after immunization with J-5 LPS with unequivocal results in protection experiments (reviewed in Müller-Loennies, et al 22 ).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of a LPS-based glycoconjugate vaccine against bovineEscherichia colimastitis: Formation of LPS Abs in cows after immunization withE. colicore oligosaccharides conjugated to hemocyanine

2012

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The immune response of cows against the core oligosaccharide of Escherichia coli rough mutants (core types R1-R4, K-12 and J-5) was investigated after immunization with a synthetic glycoconjugate composed of deacylated LPS conjugated to hemocyanine (22 animals). Ab formation was measured by ELISA using LPS or deacylated LPS conjugated to BSA as an Ag. The glycoconjugate immunogens were used to vaccinate cows (36 animals), which were then challenged intramammarily with E. coli O 157 (K1 negative, R1 core type). Compared with control groups no protection was observed, although high titers against the R1 core type were detected in vaccinated animals. Western blots using the immune sera showed that the Ab response was directed against the core region and not against the O-antigen, which may explain the failure of the vaccine.

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A novel strategy for the synthesis of neoglycoconjugates from deacylated deep rough lipopolysaccharides

Müller‐Loennies

Grimmecke

Brade

et al. 2002

Journal of Endotoxin Research

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We report a novel strategy for the preparation of neoglycoconjugates of oligosaccharides which are obtained after complete deacylation of bacterial deep rough lipopolysaccharides (LPS) isolated from recombinant Escherichia coli bacteria synthesizing a Kdo di-[alpha-Kdo-(2-->4)-alpha-Kdo-(2-->] and a Kdo trisaccharide [alpha-Kdo-(2-->8)-alpha-Kdo-(2-->4)-alpha-Kdo-(2-->] of Re-type and chlamydial LPS, respectively. Unlike acylated LPS, such oligosaccharides can be obtained in pure form and thus lead to well-defined neoglycoconjugates. Cleavage of the 1-phosphate of the lipid A moiety by alkaline phosphatase treatment leads to a free reducing glucosamine which can be further reacted with allylamine. After reductive amination, spacer elongation of the allyl group with cysteamine and activation with thiophosgene, the ligands were reacted with BSA. We have compared the immunological reactivity of such defined neoglycoconjugates obtained from natural sources with those obtained by chemical synthesis and report that such neoglycoconjugates are immunogenic and well suited as antigens for the study of epitope specificities of monoclonal antibodies. In addition, we have compared these conjugates with those in which ligands were coupled by glutardialdehyde to BSA. Our approach proved to be superior since the latter led upon immunization of mice to a relatively high percentage of antibodies that reacted with glutardialdehyde derivatized BSA without the carbohydrate ligand. This was not the case for cysteamine-spacered ligands coupled via their isothiocyanate-derivatives.

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Conserved Epitopes in Bacterial Lipopolysaccharides and Cross‐Reactive Antibodies in the Treatment of Endotoxicosis and Gram‐Negative Septic Shock

Cited by 5 publications

References 159 publications

Structural analysis of deacylated lipopolysaccharide of Escherichia coli strains 2513 (R4 core‐type) and F653 (R3 core‐type)

Structural analysis of deacylated lipopolysaccharide of Escherichia coli strains 2513 (R4 core‐type) and F653 (R3 core‐type)

Evaluation of a LPS-based glycoconjugate vaccine against bovineEscherichia colimastitis: Formation of LPS Abs in cows after immunization withE. colicore oligosaccharides conjugated to hemocyanine

A novel strategy for the synthesis of neoglycoconjugates from deacylated deep rough lipopolysaccharides

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