Influence of fine structure of lipid A on Limulus amebocyte lysate clotting and toxic activities

Takayama, K.; Qureshi, Nilofer; Raetz, C. R. H.; Ribi, Edgar; Peterson, John; Cantrell, John L.; Pearson, Frederick C.; Wiggins, Jason F.; Johnson, Arthur G.

doi:10.1128/iai.45.2.350-355.1984

Cited by 101 publications

(38 citation statements)

References 25 publications

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“…The 4'-monophosphate analog of compound 516, compound 514, was weaker than compound 516 in all of the assay systems, except immunoadjuvant activity against BSA in mice, which was examined with liposomes as the administration vehicle. This finding is consistent with our previous observation that synthetic lipid A analogs of compound series 400 and 500, lacking a phosphate group at the C-1 position (compounds 404 and 504), exhibited weaker activities than the respective 1,4'-bisphosphate compound (18,20,35) and is in accord with the observations by Qureshi et al (28) and Takayama et al (36) with bacterial products, i.e., that 4'-monophosphoryl lipid A isolated from the lipid A fraction of Salmonella typhimurium, whose chemical structure corresponds to that of compound 504, was nontoxic. The lower biological activity of R595 lipid A than compound 506 may also be explained by the fact that this bacterial product contains compounds of 4'-monophosphate structure.…”

Section: Resultssupporting

confidence: 94%

Low endotoxic activities of synthetic Salmonella-type lipid A with an additional acyloxyacyl group on the 2-amino group of beta (1-6) glucosamine disaccharide 1,4'-bisphosphate

Kotani

Takada²,

Takahashi

et al. 1986

Infect Immun

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A synthetic lipid A (Salmonella type, compound 516), P(1-6)-linked D-glucosamine disaccharide 1,4'bisphosphate, with three acyloxyacyl groups and one hydroxyacyl group, i.e., (R)-3-hexadecanoyloxytetradecanoyl, (R)-3-hydroxytetradecanoyl, (R)-3-dodecanoyloxytetradecanoyl, and (R)-3-tetradecanoyloxytetradecanoyl groups at the 2-amino, 3-hydroxyl, 2'-amino, and 3'-hydroxyl groups, respectively, was less biologically active than the synthetic Escherichia coli-type lipid A (compound 506), which has only two acyloxyacyl groups at the 2' and 3' positions and is substituted with a (R)-3-hydroxytetradecanoyl group at the 2-amino group. Compound 516 exhibited considerably weaker pyrogenic and leukopenic activity than compound 506, and it scarcely prepared rabbit skin for the Shwartzman reaction and lacked lethal toxicity on chicken embryos, although its lethal toxicity in galactosamine-loaded mice was as strong as that of compound 506. Compound 516 was also less active than compound 506 or natural E. coli lipid A (from Restrain F515) in other biological test systems, such as the Limulus test, stimulation of macrophages and lymphocytes, and interferon-inducing activity but not for interleukin-1 induction or complement activation. This observation suggests that there is an optimal number of acyloxyacyl groups on the glucosamine backbone for producing the biological activities of lipid A, especially the endotoxic activities. The 4'-monophosphate analog (compound 514) of compound 516 in general had significantly weaker activity than compound 516 in the above assays, most probably because of its greater hydrophobicity and consequently lower solubility in assay systems. Bacterial R595 lipid A derived from S. minnesota Re-mutant, which is a mixture of compounds 516 and 506, their 4'-monophosphate analogs and other compounds, exerted intermediate degrees of activity between compounds 506 and 516 in the various test systems employed. We (20) previously reported the synthesis of a lipid A with biological activities -comparable to those of a natural lipid A, which is responsible for the endotoxic and most of the other biological activities of bacterial lipopolysaccharides (LPS) (7,21,38). A synthetic compound (compound 506), P(1-6)linked D-glucosamine disaccharide 1,4'-bisphosphate, having two (R)-3-hydroxytetradecanoyl groups, the (R)-3dodecanoyloxytetradecanoyl, and the (R)-3-tetradecanoyloxytetradecanoyl group at 2-amino, 3-hydroxyl, 2'-amino and 3'-hydroxyl groups, respectively, which was prepared according to the proposed structure of lipid A from an Escherichia coli Re-mutant (F515) (10, 11; Fig. 1A), exhibited full biological activities identical to or sometimes stronger than those of the reference natural products, F515 lipid A and E. coli 055:B5 LPS.

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Section: Resultssupporting

confidence: 94%

Low endotoxic activities of synthetic Salmonella-type lipid A with an additional acyloxyacyl group on the 2-amino group of beta (1-6) glucosamine disaccharide 1,4'-bisphosphate

Kotani

Takada²,

Takahashi

et al. 1986

Infect Immun

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“…Cyanobacteria contain not only different amounts of LPS, but also their endotoxin activity differs substantially. Further research should therefore focus on the structural variability of cyanobacterial LPS, particularly lipid A, which is the major component responsible for endotoxin activities in the LAL-test (Takayama et al, 1984).…”

Section: Discussionmentioning

confidence: 99%

Isolation and endotoxin activities of lipopolysaccharides from cyanobacterial cultures and complex water blooms and comparison with the effects of heterotrophic bacteria and green alga

Bernardová

Babica

Maršálek

et al. 2007

J of Applied Toxicology

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Massive cyanobacterial water blooms are serious environmental and health problems worldwide. While some cyanobacterial toxins such as peptide microcystins have been investigated extensively, other toxic components of cyanobacteria (e.g. lipopolysaccharides, LPS) are poorly understood. The present study characterized endotoxin activities of LPS isolated from (i) laboratory cyanobacterial cultures, (ii) cyanobacterial water bloom samples dominated by Microcystis sp., Planktothrix sp., Aphanizomenon sp. and Anabaena sp., (iii) heterotrophic Gram-negative bacteria Escherichia coli, Kluyvera intermedia, Pseudomonas putida and Pseudomonas fluorescens and (iv) green alga Pseudokirchneriella subcapitata. Toxicity results derived with Limulus amebocyte lysate assay (LAL-test) showed that endotoxin activities of LPS from both cyanobacteria and heterotrophic bacteria were comparable and the values were within a similar range (1 x 10(3)-1 x 10(6) Endotoxin Units, EU, per mg of isolated LPS). The highest activities among the cyanobacterial samples were observed in the Aphanizomenon sp. dominated water bloom. The results also suggest generally higher endotoxin activities in complex natural samples than in laboratory cyanobacterial cultures. Further, experiments with the eukaryotic green alga P. subcapitata demonstrated a need for careful purification of the LPS extracts prior to testing with the LAL assay as several contaminants may overestimate endotoxin activities. This study shows relatively high pyrogenicity of LPS from various cyanobacteria. Further research should focus on detailed toxicological and ecotoxicological characterization of LPS in massive cyanobacterial water blooms.

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“…The gelation times of 5, 10 and 20 pg ml −1 MPLA were 42·6, 32·3 and 25·9 min, respectively, and the activity ratio of MPLA relative to lipid A was 23·1% (Table ), which was lower than that previously reported (Takayama et al . ; Johnson et al . ).…”

Section: Resultsmentioning

confidence: 99%

Iron sulfate inhibits Limulus activity by induction of structural and qualitative changes in lipid A

Fujita¹,

Nabetani²

2013

J Appl Microbiol

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Aims: The bacterial endotoxins test (BET) is a sensitive assay for measuring endotoxin levels in solution and uses the limulus amebocyte lysate (LAL) coagulation reaction. We sought to identify the mechanisms through which certain substances interfere with the interaction between LAL and bacterial lipopolysaccharide (LPS). Methods and Results: Endotoxin lipid A was inactivated by the addition of iron sulfate, which acted on endotoxin directly and strongly inhibited LAL coagulation activity. Size-exclusion, anion-exchange and reverse-phase liquid chromatography/mass spectrometry were used to examine changes in inactivated lipid A in terms of complex formation, negative charge status, hydrophobic interaction and structure. Furthermore, we verified the involvement of the lipid A phosphoryl group in the interference of iron sulfate with lipid A-factor C binding activity. Iron sulfate-inactivated lipid A was cleaved at its glycosidic bond, resulting in loss of hydrophobic interactions and disruption of lipid A complexes without alteration of negative charge status and lipid A-factor C interaction. Conclusions: Lipid A cleavage was a direct result of interfering factors, including iron sulfate, which acted on endotoxin directly to disrupt lipid A complexes rather than interfering with LAL coagulation. Significance and Impact of the Study: Our data provide new insights into the mechanisms of lipid A activity.

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Influence of fine structure of lipid A on Limulus amebocyte lysate clotting and toxic activities

Cited by 101 publications

References 25 publications

Low endotoxic activities of synthetic Salmonella-type lipid A with an additional acyloxyacyl group on the 2-amino group of beta (1-6) glucosamine disaccharide 1,4'-bisphosphate

Low endotoxic activities of synthetic Salmonella-type lipid A with an additional acyloxyacyl group on the 2-amino group of beta (1-6) glucosamine disaccharide 1,4'-bisphosphate

Isolation and endotoxin activities of lipopolysaccharides from cyanobacterial cultures and complex water blooms and comparison with the effects of heterotrophic bacteria and green alga

Iron sulfate inhibits Limulus activity by induction of structural and qualitative changes in lipid A

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