Structure-Activity Relationship of Synthetic Toll-like Receptor 4 Agonists

Stöver, Axel G.; Correia, Jean da Silva; Evans, John K.; Cluff, C W; Elliott, Mark; Jeffery, Eric; Johnson, David A.; Lacy, Michael J.; Baldridge, Jory R.; Probst, Peter; Ulevitch, Richard J.; Persing, David H.; Hershberg, Robert M.

doi:10.1074/jbc.m310760200

Cited by 150 publications

(148 citation statements)

References 36 publications

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“…Differences in lipid A structure or conformation are known to influence biological activity by changing the supramolecular conformation of lipid A [22][23][24][25]. A bisphosphorylated, C12 and C14, hexa-acylated lipid A structure linked to KDO is the most bioactive when TNFα is the measure of bioactivity [24,26,27].…”

Section: Discussionmentioning

confidence: 99%

TLR4-dependent adjuvant activity of Neisseria meningitidis lipid A

Zughaier

Steeghs

Ley³

et al. 2007

Vaccine

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The adjuvant activity of Neisseria meningitidis serogroup B lipopoly(oligo)saccharide (LOS) from wild-type and genetically-defined LOS mutants and unglycosylated meningococcal lipid A was assessed in C3H/HeN and C3H/HeJ mice. Meningococcal lipid A, a weak agonist for TLR4/MD-2 in human macrophages, was found to have adjuvant activity similar to that of wild-type and KDO 2 -lipid A LOS in C3H/HeN mice. All meningococcal LOS structures as adjuvants induced high titers of IgG1, IgG2a and IgG2b but very little IgG3 to OMP compared to no adjuvant PBS controls. In addition, induced OMP antibodies were shown to have high bactericidal activity against serogroup B meningococci. Purified LOS and lipid A structures failed to induce any adjuvant activity in C3H/ HeJ mice indicating that meningococcal LOS as an adjuvant was TLR4-dependent. Unglycosylated meningococcal lipid A because of its weak agonist activity for human macrophages and retention of adjuvant activity may be a candidate for use in serogroup B meningococcal OMP and OMV vaccines and for use as an adjuvant in other vaccines.

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Section: Discussionmentioning

confidence: 99%

TLR4-dependent adjuvant activity of Neisseria meningitidis lipid A

Zughaier

Steeghs

Ley³

et al. 2007

Vaccine

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“…One study demonstrated that the number of phosphates and the number and length of the acyl chains appended to LPS molecules dictate whether they will bind TLR2 or TLR4 (7). Another study observed that a mutant Escherichia coli strain produced a penta-acylated LPS molecule that was considerably less stimulatory when compared with the well known hexaacylated structure (30), and yet others have demonstrated that the presence of a secondary acyl chain length of 10 carbons was optimal for hexa-acylated LPS to induce a TLR4-dependent pro-inflammatory response (24).…”

Section: Discussionmentioning

confidence: 99%

“…The structure of GPLs makes them unique TLR2 agonists since they essentially constitute a "mixture" of glycolipids and lipopeptides. Moreover, they contain only one N-linked fatty acid chain, thereby differing from other TLR2 (and TLR4) ligands, which usually contain multiple fatty acids of varying length that can be either O-linked or N-linked (7,(22)(23)(24). In the present study, we show that specific acetylation and methylation patterns of the 6-deoxytalose and rhamnose, respectively, which comprise the diglycosylated lipopeptide core of GPL, are required for GPL signaling through the macrophage TLR2.…”

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confidence: 99%

Mycobacterium avium Glycopeptidolipids Require Specific Acetylation and Methylation Patterns for Signaling through Toll-like Receptor 2

Sweet

Zhang

Torres-Fewell

et al. 2008

Journal of Biological Chemistry

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Toll-like receptors (TLRs) recognize pathogen-associated molecules and play a vital role in promoting an immune response against invading microbes. TLR2, one of the key members of the TLR family, recognizes a wide variety of microbial products, including lipoproteins and lipopeptides, from a number of pathogens. Recent studies from our laboratory indicate that glycopeptidolipids (GPLs), a major surface component of Mycobacterium avium and other non-tuberculosis mycobacteria, are ligands for TLR2. However, the molecular requirements necessary for the GPL-TLR2 interaction were not defined in this report. In the present study we isolated different GPL species from M. avium, and using mass spectrometry and NMR analyses, characterized the molecular requirements of the GPL-TLR2 interaction. Interestingly, the extent of the respective acetylation and methylation of the 6-deoxytalose and rhamnose contained within the core GPL structure dictated whether the GPL signaled through TLR2. These experiments illustrate how subtle changes in a complex TLR2 ligand can alter its affinity for this important receptor, and suggest that M. avium could potentially modify its GPL structure to limit its interaction with TLR2.

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“…Ultra-pure LPS (from Escherichia coli serotype 0111:B4) was purchased from InvivoGen. CRX-527 was synthesized and provided by GlaxoSmithKline Biologicals [5]. C57BL/6 WT and CD14-deficient mice were obtained from Harlan and Charles River Laboratories, respectively, and were bred and maintained in specific pathogen-free conditions according to institutional guidelines.…”

Section: Reagents Cell Lines and Animalsmentioning

confidence: 99%

“…The hexa-acylate compound CRX-527, composed of three myristic and three decanoic acyl chains, has been described as one of the most powerful members of this aminoalkyl glucosaminide 4-phosphate family [5]. CD14, a GPI-anchored protein expressed at the surface of phagocytes, has been shown to be involved in the induction of TLR4-mediated responses by bacterial LPS [6,7].…”

Section: Introductionmentioning

confidence: 99%

CD14‐independent responses induced by a synthetic lipid A mimetic

et al. 2010

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CRX-527 belongs to a new family of synthetic lipid A mimetics, the aminoalkyl glucosaminide 4-phosphates, which are considered as potential vaccine adjuvants or standalone immunotherapeutics to harness innate immune defenses. Since natural lipid A from bacterial LPS depends on membrane-bound (mCD14) or soluble CD14 for its TLR4 ligand activity, we investigated the involvement of both forms of CD14 in the responses elicited by CRX-527. First, we found that CRX-527 induces NF-jB and interferon regulatory factor-3 (IRF-3) activation in human embryonic kidney cells transfected with TLR4 and MD-2 genes alone, whereas the responses to LPS require either co-transfection of the gene encoding mCD14 or addition of soluble CD14. We then observed that monocyte-derived DC, which are devoid of mCD14 respond to CRX-527 but not to LPS in serum-free medium. Furthermore, we found that, in contrast to LPS, CRX-527 induces the production of cytokines in whole blood of a patient with paroxysmal nocturnal hemoglobinuria, a disease in which mCD14-dependent responses are defective. Finally, we demonstrated that splenocytes from CD14-deficient mice produce cytokines in response to CRX-527 but not to LPS. We conclude that the lipid A mimetic CRX-527 does not require the CD14 co-receptor to elicit TLR4-mediated responses.Key words: CD14 . Lipid A . TLR4 Supporting Information available online IntroductionAminoalkyl glucosaminide phosphates lipid A mimetics are currently developed as synthetic Toll-like receptor 4 (TLR4) agonists with possible applications as vaccine adjuvants or standalone immunostimulants [1][2][3][4]. The hexa-acylate compound CRX-527, composed of three myristic and three decanoic acyl chains, has been described as one of the most powerful members of this aminoalkyl glucosaminide 4-phosphate family [5]. CD14, a GPI-anchored protein expressed at the surface of phagocytes, has been shown to be involved in the induction of TLR4-mediated responses by bacterial LPS [6,7]. Indeed, LPS was shown to bind the LPS binding protein facilitating its transfer to membranebound CD14 (mCD14), which allows LPS to engage TLR4 associated with MD-2 [6]. There is evidence that CD14 is especially important for TLR4 response mediated by the TRIF adaptor molecule, which culminates in activation of the transcription factor interferon regulatory factor-3 (IRF3) required for the production of type I interferon [8]. Cells which express TLR4 but not mCD14, such as endothelial cells and monocytederived DC (MoDC), can use the soluble form of CD14 (sCD14) to mount TLR4-mediated responses to LPS [9]. sCD14 was recently shown to discriminate slight structural differences between distinct TLR4 ligands [10]. The present study was therefore SHORT COMMUNICATION Results and discussionExpression of TLR4 and MD-2 is sufficient for the induction of NF-jB and IRF3 activities by CRX-527In preliminary experiments using transfected human embryonic kidney (HEK) cells expressing different TLR receptors, we confirmed that CRX-527 is a TLR4 agonist devoid of activity o...

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Structure-Activity Relationship of Synthetic Toll-like Receptor 4 Agonists

Cited by 150 publications

References 36 publications

TLR4-dependent adjuvant activity of Neisseria meningitidis lipid A

TLR4-dependent adjuvant activity of Neisseria meningitidis lipid A

Mycobacterium avium Glycopeptidolipids Require Specific Acetylation and Methylation Patterns for Signaling through Toll-like Receptor 2

CD14‐independent responses induced by a synthetic lipid A mimetic

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