Endotoxic properties of chemically synthesized lipid A part structures. Comparison of synthetic lipid A precursor and synthetic analogues with biosynthetic lipid A precursor and free lipid A

Galanos, Chris; Lehmann, Volker; Lüderitz, Otto; Rietschel, Ernst; Westphal, Otto; Brade, Helmut; Brade, Lore; Freudenberg, Marina A.; Hansen‐Hagge, Thomas E.; Lüderitz, Thomas; McKENZIE, Gerry; Schade, Ulrich; Strittmatter, Wolfgang; Tanamoto, Ken‐ichi; Zähringer, Ulrich; Imoto, Masahiro; Yoshimura, Hidetoshi; Yamamoto, Misa; Shimamoto, Takuya; Kusumoto, Shoichi; Shiba, Tetsuo

doi:10.1111/j.1432-1033.1984.tb08090.x

Cited by 170 publications

(88 citation statements)

References 20 publications

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“…First, the lipid A from Salmonella LPS is specifically inactive on human cells, showing that recognition of the lipid A structure by human cells is different from that of mice or rabbits. The same species-specific phenomenon had been observed with tetraacylated lipid A precursor (lipid IV A ), which is also active in mice and rabbits, but inactive on human cells (9,33). However, the meaning of these two lipid As is completely different in view of the fact that the precursor structure is an intermediate in the biosynthesis of LPS, whose structure is not found as a component of any LPS.…”

Section: Discussionmentioning

confidence: 84%

Salmonella-Type Heptaacylated Lipid A Is Inactive and Acts as an Antagonist of Lipopolysaccharide Action on Human Line Cells

Tanamoto

Azumi

2000

The Journal of Immunology

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The stimulation of both THP-1 and U937 human-derived cells by Salmonella lipid A preparations from various strains, as assessed by TNF-α induction and NF-κB activation, was found to be very low (almost inactive) compared with Escherichia coli lipid A, but all of the lipid As exerted strong activity on mouse cells and on Limulus gelation activity. Experiments using chemically synthesized E. coli-type hexaacylated lipid A (506) and Salmonella-type heptaacylated lipid A (516) yielded clearer results. Both lipid A preparations strongly induced TNF-α release and activated NF-κB in mouse peritoneal macrophages and mouse macrophage-like cell line J774-1 and induced Limulus gelation activity, although the activity of the latter was slightly weaker than that of the former. However, 516 was completely inactive on both THP-1 and U937 cells in terms of both induction of TNF-α and NF-κB activation, whereas 506 displayed strong activity on both cells, the same as natural E. coli LPS. In contrast to the action of the lipid A preparations, all the Salmonella LPSs also exhibited full activity on human cells. However, the polysaccharide portion of the LPS neither exhibited TNF-α induction activity on the cells when administered alone or together with lipid A nor inhibited the activity of the LPS. These results suggest that the mechanism of activation by LPS or the recognition of lipid A structure by human and mouse cells may differ. In addition, both 516 and lipid A from Salmonella were found to antagonize the 506 and E. coli LPS action that induced TNF-α release and NF-κB activation in THP-1 cells.

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Section: Discussionmentioning

confidence: 84%

Salmonella-Type Heptaacylated Lipid A Is Inactive and Acts as an Antagonist of Lipopolysaccharide Action on Human Line Cells

Tanamoto

Azumi

2000

The Journal of Immunology

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“…Thus, all structural elements required for the induction of these endotoxin activities are present in the lipid A precursor indicating that for these effects 3-acyloxyacyl groups are not important. The bacterial and synthetic precursor molecules were, however, found to be of weak pyrogenicity and to be incapable of preparing for or eliciting the local Shwartzman reaction [30,311. This suggests that the presence of 3-acyloxyacyl groups is necessary for the full expression of these endotoxic reactions.…”

Section: Discussionmentioning

confidence: 99%

“…This structure has recently been chemically synthesized [29] and been analysed biologically in comparison with the natural lipid A precursor and bacterial free lipid A in various endotoxin tests. It was found that the bacterial and synthetic precursor exhibit strong activities in a number of systems in vivo and in vitro including lethal toxicity, B-lymphocyte mitogenicity and the capacity to activate macrophages [30,311. Thus, all structural elements required for the induction of these endotoxin activities are present in the lipid A precursor indicating that for these effects 3-acyloxyacyl groups are not important.…”

Section: Discussionmentioning

confidence: 99%

Nature and location of amide‐bound (R)‐3‐acyloxyacyl groups in lipid A of lipopolysaccharides from various gram‐negative bacteria

Wollenweber

Seydel

Lindner

et al. 1984

European Journal of Biochemistry

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It has previously been demonstrated [Eur. J. Biochem. 124, 191-198 (1982) and 137, 15-22 (198311 that the lipid A component of Salmonella and Proteus lipopolysaccharides contains amide-linked (R)-3-acyloxyacyl residues. In the present study lipid A of other gram-negative bacteria was analysed for the presence of amidebound 3-acyloxyacyl residues. It was found that such residues are constituents of all lipid A tested (Agrobacterium tumefaciens, Chromobacterium violaceum, Pseudomonas aeruginosa, Xanthomonas sinensis, Bacteriodes frugilis, Vibrio cholerae, Fusobacterium nucleatum, Rhodospirillum tenue, Acinetobacter calcoaceticus, and Escherichia coli). Amide-linked (R)-3-acyloxyacyl groups, therefore, represent common and ubiquitous structural elements of bacterial lipid A.The composition of 3-acyloxyacyl groups differed considerably among different bacteria. As amide-bound (R)-3-hydroxy fatty acids straight chain and isobranched acyl groups with 10-17 carbon atoms were identified. The most frequently encountered fatty acids, substituting the 3-hydroxyl group of 3-hydroxy fatty acids, were nonhydroxylated straight chain and isobranched acyl residues with 10-17 carbon atoms as well as (Q-2-hydroxy fatty acids with 12 carbon atoms.In some cases, using laser desorption mass spectrometry, the distribution of 3-acyloxyacyl residues over the two available glucosamine amino groups of the lipid A backbone was investigated.Lipid A, the covalently bound lipid component of bacterial lipopolysaccharides (endotoxins) is, in general, made up by a bisphosphorylated p1 ,6-linked D-glucosamine disaccharide (lipid A backbone) which carries up to 4 mol of long-chain (R)-3-hydroxy fatty acid residues [I]. As was demonstrated for lipid A of Salmonella minnesota, Salmonella typhimurium, Escherichia coli and Proteus mirabilis 2 mol of these are linked to amino groups (positions 2 and 2') and 2 mol are bound to hydroxyl groups at positions 3 and 3' of the lipid A backbone 11-41. It has also previously been demonstrated that in these bacteria approximately 50% of the ester-linked (R)-3-hydroxy fatty acids are 3-0-acylated by straight-chain nonhydroxylated or (S)-Zhydroxy fatty acids [l, 3-61. Very recently it was shown for enterobacterial lipid A that also the amidebound (R)-3-hydroxy fatty acids (notably that at position 2') carry nonhydroxylated fatty acids at their 3-hydroxyl group in a specific distribution [7, 81. From these results it appeared that (R)-3-acyloxyacyl groups are important and dominating lipophilic constituents of lipid A and it was speculated that they may be also present in lipid A of bacteria other than Enterobacteriaceae.The studies reported in this paper, therefore, were performed to analyse lipid A of various bacterial origin for the presence and location of amide-linked (R)-3-acyloxyacyl residues. It will be shown that in all lipid A samples studied 3-acyloxyacyl groups are present indicating that they are common and prominent constituents of bacterial lipid A.Abbreviations. The abbreviations for fatty acids fo...

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“…The revised structure was based on the studies by high resolution mass spectrometry and two-dimensional nuclear magnetic resonance [I 3 -151. The newly synthesized analogues were mainly distributed to three groups in Tokyo (this study), Osaka [25] and Freiburg [26] and each group independently investigated biological activities, some common to the groups but some not.…”

Section: Discussionmentioning

confidence: 99%

Biological activities of analogues of lipid A based chemically on the revised structural model

Kanegasaki

Kojima

Matsuura

et al. 1984

European Journal of Biochemistry

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Lipid A analogues were chemically synthesized based on the model structure recently revised, and biological activities of the analogues were tested. The analogue, (p-I,6)-linked glucosamine disaccharide carrying ester-bound 3-hydroxytetradecanoic acids at 3 and 3' position of reducing and nonreducing glucosamine in addition to amidebound 3-hydroxytetradecanoic acids and glycosidic-linked and ester-linked phosphate groups, showed much stronger activities for mediator inducing and immunomodulating as well as endotoxic activities than those exhibited by the previously synthesized analogues based on the old model. Among the activities tested, induction of interferon and tumor necrosis factor as well as mitogenicity, adjuvanticity and pyrogenicity were, however, not expressed so strongly as natural lipid A used as controls. In contrast, the analogue exhibited comparable activities to those of control lipid A in the test of lethal toxicity to mice and gelating activity of Limulus amebocyte lysate. Other synthetic analogues carrying a phosphate group showed comparable, slightly stronger or weaker activities depending on the test, but nonphosphorylated analogue exhibited no apparent or only very weak activities.It is well established that lipid A component of lipopolysaccharides of gram-negative bacteria is responsible for most, if not all, various biological and endotoxic activities including lethal toxicity, pyrogenicity, immunomodulating activities, and the activities to induce various mediators [I]. The chemical structure of the lipid A backbone in many bacteria appears to be identical and consists of a (/3-1,6)-linked D-glucosamine disaccharide which carries phosphate residues in the positions of 1 and 4 as well as amide-bound and ester-bound D-3-hydroxy and/or acyloxy fatty acids [2], although a number of minor differences in structure are known in lipid A derived from various bacteria. It is also known that isolated free lipid A is a mixture of congeners.To confirm lipid A structure and to establish the structure responsible for the functions, compounds based on the proposed structure were recently chemically synthesized [3 -51 and their biological activities were investigated in many laboratories [6-121. Although some of the synthetic analogues have been reported to show significant biological activities such as interferon inducing [7] as well as mitogenic and adjuvant activities [6, 7, 1 I], the activity of the analogues, if any [6-121 were much weaker than natural lipid A.More recently, locations of ester-linked 3-hydroxytetradecanoic acid (or acyloxy fatty acid) of lipid A from Escherichia coli, Salmonella typhimurium and Proteus mirahilis were reinvestigated. The results disclosed that the locations were not at the 3,4 and 6' positions as presumed previously but at the 3 and 3' positions of diglucosamine unit 113-151. The compound with such altered structure, therefore, was hoped to enhance biological activities and represent comparable activities to natural lipid A. Based on the modified structure, analogue...

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Endotoxic properties of chemically synthesized lipid A part structures. Comparison of synthetic lipid A precursor and synthetic analogues with biosynthetic lipid A precursor and free lipid A

Cited by 170 publications

References 20 publications

Salmonella-Type Heptaacylated Lipid A Is Inactive and Acts as an Antagonist of Lipopolysaccharide Action on Human Line Cells

Salmonella-Type Heptaacylated Lipid A Is Inactive and Acts as an Antagonist of Lipopolysaccharide Action on Human Line Cells

Nature and location of amide‐bound (R)‐3‐acyloxyacyl groups in lipid A of lipopolysaccharides from various gram‐negative bacteria

Biological activities of analogues of lipid A based chemically on the revised structural model

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