Synthesis of the First Fully Active Lipoteichoic Acid

Stadelmaier, Andreas; Morath, Siegfried; Hartung, Thomas; Schmidt, Richard R.

doi:10.1002/anie.200390243

Cited by 52 publications

(35 citation statements)

References 25 publications

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“…LTA from Staphylococcus aureus (DSM 20233) was prepared by n-butanol extraction, as described previously (8). The general strategy of Staphylococcus aureus LTA synthesis has been published elsewhere (15), and details on the synthesis of the structural analogs investigated in this paper are also reported elsewhere (2). LPS from Salmonella enterica serovar Abortus Equi was purchased from Sigma (Deisenhofen, Germany), and recombinant human tumor necrosis factor alpha (TNF-␣) was obtained from Innogenetics (Heiden, Germany).…”

Section: Methodsmentioning

confidence: 99%

Use of Synthetic Derivatives To Determine the Minimal Active Structure of Cytokine-Inducing Lipoteichoic Acid

Deininger

Figueroa-Perez

Sigel

et al. 2007

Clin Vaccine Immunol

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Lipoteichoic acid (LTA) from gram-positive bacteria is the counterpart to lipopolysaccharide from gramnegative bacteria. LTA, which activates Toll-like receptor 2 (TLR2), induces a unique cytokine and chemokine pattern. The chemical synthesis of LTA proved its immunostimulatory properties. To determine the minimal active structure of LTA, we reduced synthetic LTA in a number of steps down to the synthetic anchor and employed these molecules to stimulate interleukin-8 (IL-8) release in human whole blood. Ten times more of the synthetic structures with four to six D-alanine-substituted polyglycerophosphate units (50 nM) than of the native LTA preparation was required to induce IL-8 release. A further reduction to three backbone units with two or no D-alanine residues resulted in cytokine induction only from 500 nM. The synthetic anchor was not able to induce IL-8 release even at 5 M. When the LTA derivatives were used at 500 nM, they induced increasing levels of IL-8 and tumor necrosis factor alpha with increasing elongation of the backbone. Peritoneal macrophages were less responsive than human blood to the synthetic structures. Therefore, TLR2 dependency could be shown only with cells from TLR2-deficient mice for the two largest synthetic structures. This was confirmed by using TLR2-transfected HEK 293 cells. Taken together, these data indicate that although the synthetic anchor (which, unlike the native anchor, contains only myristic acid) cannot induce cytokine release, the addition of three backbone units, even without D-alanine substituents, confers this ability. Lengthening of the chain with D-alanine-substituted backbone units results in increased cytokine-inducing potency and a more sensitive response.Recognition of conserved bacterial structures called pathogen-associated molecular patterns occurs via pattern recognition receptors on immune cells and leads to activation of the innate immune system and the induction of a variety of cytokines. Lipopolysaccharide (LPS) has been known as the most important pathogen-associated molecular pattern of gramnegative bacteria for more than 50 years (16) and has been well examined in detail over the decades. Immune recognition takes place by the binding of LPS to Toll-like receptor 4 (TLR4) and also involves the cofactors CD14 (17) and 14).The immunostimulatory component of gram-positive bacteria was not clear for a long time, although a structural counterpart to LPS, called lipoteichoic acid (LTA), was found in the bacterial membrane. Like LPS, LTA is an amphiphilic molecule with a lipid anchor and a negatively charged backbone. Inefficient preparation methods on the basis of hot phenol, which resulted in the decomposition and the subsequent loss of activity, or LPS contamination during preparation led to inconsistent findings (9). Meanwhile, an improved preparation method based on n-butanol extraction at an ambient temperature was developed to purify the biologically active LTA of Staphylococcus aureus (8) and other organisms (3, 4). Structural analysis by nuclear ...

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Section: Methodsmentioning

confidence: 99%

Use of Synthetic Derivatives To Determine the Minimal Active Structure of Cytokine-Inducing Lipoteichoic Acid

Deininger

Figueroa-Perez

Sigel

et al. 2007

Clin Vaccine Immunol

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“…S. aureus LTA is composed of poly-Gro-P, on average 30-50 residues, with D-ala and N-acetylglucosamine (GlcNAc) as major substituents. A synthetic analogue of this LTA, (D-ala) 4 (GlcNAc) 1 (Gro-P) 6 gentiobiosyl-sn-dimyristoylglycerol, was shown to induce cytokine release with the same potency as native LTA [21,22]. For maximal activity, both the glycolipid and D-ala residues were required, whereas GlcNAc substituents could be omitted.…”

Section: Probiotics and Immunomodulationmentioning

confidence: 99%

Anti-inflammatory potential of probiotics: lipoteichoic acid makes a difference

Lebeer

Claes

Vanderleyden

2012

Trends in Microbiology

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“…The detailed synthesis has been published (22). The identities of the molecules were characterized by matrixassisted laser desorption-ionization time-of-flight mass spectrometry and nuclear magnetic resonance (NMR) at 600.13 MHz and 300 K. The NMR spectra were related to 3-(trimethysilyl) 3,3,2,2-tetradeuteropropionic acid Na salt (d 4 -TSPA).…”

Section: Stimulimentioning

confidence: 99%

Definition of Structural Prerequisites for Lipoteichoic Acid-Inducible Cytokine Induction by Synthetic Derivatives

Deininger

Stadelmaier

Aulock

et al. 2003

The Journal of Immunology

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The controversy about the immune stimulatory properties of lipoteichoic acid (LTA) from Staphylococcus aureus was solved recently by showing decomposition and inactivation of LTA obtained by conventional purification strategies, as well as pronounced LPS contamination of commercial preparations. By introducing a novel preparation method, the structure of bioactive LTA was elucidated. This structure was confirmed by chemical synthesis. In this work, synthetic LTA derivatives were employed to study the structure-function relationship of cytokine induction in human monocytes. Synthetic LTA induced the same cytokine pattern as highly purified natural LTA. The gentiobiose core could be omitted without affecting bioactivity. The polyglycerophosphate backbone amplified the response to the lipid anchor (∼100-fold) only when substituted with d-alanine, whereas α-d-N-acetylglucosamine substituents could be omitted. Replacing d-alanine substituents with l-alanine reduced the activity of the molecule at least 10-fold, indicating stereoselectivity. These results define for the first time the crucial patterns required for the immune recognition of LTA.

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Synthesis of the First Fully Active Lipoteichoic Acid

Cited by 52 publications

References 25 publications

Use of Synthetic Derivatives To Determine the Minimal Active Structure of Cytokine-Inducing Lipoteichoic Acid

Use of Synthetic Derivatives To Determine the Minimal Active Structure of Cytokine-Inducing Lipoteichoic Acid

Anti-inflammatory potential of probiotics: lipoteichoic acid makes a difference

Definition of Structural Prerequisites for Lipoteichoic Acid-Inducible Cytokine Induction by Synthetic Derivatives

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