Lipopeptides from <i>Staphylococcus aureus</i> as Tlr2 Ligands: Prediction with mRNA Expression, Chemical Synthesis, and Immunostimulatory Activities

Fujimoto, Yukari; Hashimoto, Masahito; Furuyashiki, Maiko; Katsumoto, Mami; Seya, Tsukasa; Suda, Yasuo; Fukase, Koichi

doi:10.1002/cbic.200900242

Cited by 19 publications

(24 citation statements)

References 26 publications

(35 reference statements)

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“…We synthesized 16 Pam2Cys-containing lipopeptides using S. aureus lipoproteins as a reference [30], and designated them Pam2Cys1-Pam2Cys16 (Table 1). Included as positive controls were the diacyl lipopeptides Pam2CSK4 [31], designated as Pam2CSK19 in this study, and its derivatives Pam2CSK (Pam2Cys17), and Pam2CSK2 (Pam2Cys18).…”

Section: Resultsmentioning

confidence: 99%

“…The synthesis of lipopeptides was achieved with a combination of solution- and solid-phase methods [30]. Briefly, for the preparation of the Pam2Cys backbone, the protected cysteine (Fmoc-Cys-OtBu) and the iodide (3-iodopropane-1,2-diol) were coupled under basic condition by using Cs 2 CO 3 to give Fmoc-Cys(2,3-dihydroxypropyl)-OtBu, and the subsequent acylation and cleavage of the tBu group gave Fmoc-Pam2Cys-OH [37].…”

Section: Methodsmentioning

confidence: 99%

“…Their diacylated structure was confirmed by MS/MS spectrometry [28]. We annotated these lipoproteins by function, which largely depends on their protein sequence [30]. Based on these results, we chemically synthesized 16 Pam2Cys lipopeptides of 6–18 amino acids (a.a.) [30].…”

Section: Introductionmentioning

confidence: 99%

“…We annotated these lipoproteins by function, which largely depends on their protein sequence [30]. Based on these results, we chemically synthesized 16 Pam2Cys lipopeptides of 6–18 amino acids (a.a.) [30]. They possessed TLR2 agonistic activity, but varied in their functional potential to activate NF-κB and liberate TNF-α from human PBMC [30], yet their NK activation potential has not been determined.…”

Section: Introductionmentioning

confidence: 99%

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The Peptide Sequence of Diacyl Lipopeptides Determines Dendritic Cell TLR2-Mediated NK Activation

et al. 2010

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Natural killer (NK) cells are lymphocyte effectors that are activated to control certain microbial infections and tumors. Many NK-activating and regulating receptors are involved in regulating NK cell function. In addition, activation of naïve NK cells is fundamentally triggered by cytokines or myeloid dendritic cells (mDC) in various modes. In this study, we synthesized 16 S-[2,3-bis(palmitoyl)propyl]cysteine (Pam2Cys) lipopeptides with sequences designed from lipoproteins of Staphylococcus aureus, and assessed their functional properties using mouse (C57BL/6) bone marrow-derived DC (BMDC) and NK cells. NK cell activation was evaluated by three criteria: IFN-γ production, up-regulation of NK activation markers and cytokines, and NK target (B16D8 cell) cytotoxicity. The diacylated lipopeptides acted as TLR2 ligands, inducing up-regulation of CD25/CD69/CD86, IL-6, and IL-12p40, which represent maturation of BMDC. Strikingly, the Pam2Cys lipopeptides induced mouse NK cell activation based on these criteria. Cell-cell contact by Pam2Cys peptide-stimulated BMDC and NK cells rather than soluble mediators released by stimulated BMDC induced activation of NK cells. For most lipopeptides, the BMDC TLR2/MyD88 pathway was responsible for driving NK activation, while some slightly induced direct activation of NK cells via the TLR2/MyD88 pathway in NK cells. The potential for NK activation was critically regulated by the peptide primary sequence. Hydrophobic or proline-containing sequences proximal to the N-terminal lipid moiety interfered with the ability of lipopeptides to induce BMDC-mediated NK activation. This mode of NK activation is distinctly different from that induced by polyI:C, which is closely associated with type I IFN-inducing pathways of BMDC. These results imply that the MyD88 pathway of BMDC governs an alternative NK-activating pathway in which the peptide sequence of TLR2-agonistic lipopeptides critically affects the potential for NK activation.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%