Advertisement

Amino Acids

, Volume 39, Issue 2, pp 471–480 | Cite as

Structural requirement for the agonist activity of the TLR2 ligand Pam2Cys

  • Weiguang ZengEmail author
  • Emily Eriksson
  • Brendon Chua
  • Lara Grollo
  • David C. JacksonEmail author
Original Article

Abstract

Synthetic lipopeptides have demonstrated great potential as a vaccine strategy for eliciting cellular and humoral immunity. One of the most potent lipid moieties used is S-[2,3-bis(palmitoyloxy)propyl]cysteine (Pam2Cys). Pam2Cys binds to and activates dendritic cells by engagement of Toll like receptor 2 (TLR 2). In this study, we have investigated the structural requirement of the agonist activity of Pam2Cys by varying the three structural elements of the core structure S-(2,3-dihydroxypropyl)-cysteine namely (1) the α-amino group of the cysteine residue (2) the sulphur atom of the cysteine residue and (3) the 2,3-dihydroxypropyl moiety. Four novel analogues of Pam2Cys were made and each of these analogues were incorporated into vaccine constructs and examined for immunogenicity. Our results demonstrate that (1) the potency of the peptide vaccine is least affected by removal of the amino group (2) substitution of the sulphur atom with an amide bond leads to significant reduction of biological activity (3) removal of the amino group and at the same time substitution of the sulphur with an amide bond significantly decreases the biological activity (4) in the two analogues in which the sulphur atom is replaced with an amide bond the analogue containing the 1,3-dihydroxypropyl moiety demonstrates higher activity than the one which contains 2,3-dihydroxypropyl. In conclusion, the results demonstrate strict structural requirements for agonist activity of the TLR2 ligand Pam2Cys.

Keywords

Vaccine Peptide TLR2 Structure–activity relationship Lipopeptide 

Notes

Acknowledgments

This work was supported by grant from the National Health and Medical Research Council of Australia.

References

  1. Alphs HH, Gambhira R, Karanam B, Roberts JN, Jagu S, Schiller JT, Zeng W, Jackson DC, Roden RB (2008) Protection against heterologous human papillomavirus challenge by a synthetic lipopeptide vaccine containing a broadly cross-neutralizing epitope of L2. Proc Natl Acad Sci USA 105:5850–5855CrossRefPubMedGoogle Scholar
  2. Batzloff MR, Hartas J, Zeng W, Jackson DC, Good MF (2006) Intranasal vaccination with a lipopeptide containing a conformationally constrained conserved minimal peptide, a universal T cell epitope, and a self-adjuvanting lipid protects mice from group A streptococcus challenge and reduces throat colonization. J Infect Dis 194:325–330CrossRefPubMedGoogle Scholar
  3. Baz A, Buttigieg K, Zeng W, Rizkalla M, Jackson DC, Groves P, Kelso A (2008) Branched and linear lipopeptide vaccines have different effects on primary CD4+ and CD8+ T-cell activation but induce similar tumor-protective memory CD8+ T-cell responses. Vaccine 26:2570–2579CrossRefPubMedGoogle Scholar
  4. Belz G, Smith C, Bharadwaj M, Rice A, Jackson D (2004) DCs as targets for vaccine design. Cytotherapy 6:88–98CrossRefPubMedGoogle Scholar
  5. Borges E, Wiesmuller KH, Jung G, Walden P (1994) Efficacy of synthetic vaccines in the induction of cytotoxic T lymphocytes. Comparison of the costimulating support provided by helper T cells and lipoamino acid. J Immunol Methods 173:253–263CrossRefPubMedGoogle Scholar
  6. Buwitt-Beckmann U, Heine H, Wiesmuller KH, Jung G, Brock R, Akira S, Ulmer AJ (2005a) Toll-like receptor 6-independent signaling by diacylated lipopeptides. Eur J Immunol 35:282–289CrossRefPubMedGoogle Scholar
  7. Buwitt-Beckmann U, Heine H, Wiesmuller KH, Jung G, Brock R, Ulmer AJ (2005b) Lipopeptide structure determines TLR2 dependent cell activation level. Febs J 272:6354–6364CrossRefPubMedGoogle Scholar
  8. Buwitt-Beckmann U, Heine H, Wiesmuller KH, Jung G, Brock R, Akira S, Ulmer AJ (2006) TLR1- and TLR6-independent recognition of bacterial lipopeptides. J Biol Chem 281:9049–9057CrossRefPubMedGoogle Scholar
  9. Chua BY, Zeng W, Lau YF, Jackson DC (2007) Comparison of lipopeptide-based immunocontraceptive vaccines containing different lipid groups. Vaccine 25:92–101CrossRefPubMedGoogle Scholar
  10. Day EB, Zeng W, Doherty PC, Jackson DC, Kedzierska K, Turner SJ (2007) The context of epitope presentation can influence functional quality of recalled influenza A virus-specific memory CD8+ T cells. J Immunol 179:2187–2194PubMedGoogle Scholar
  11. Deliyannis G, Kedzierska K, Lau YF, Zeng W, Turner SJ, Jackson DC, Brown LE (2006) Intranasal lipopeptide primes lung-resident memory CD8+ T cells for long-term pulmonary protection against influenza. Eur J Immunol 36:770–778CrossRefPubMedGoogle Scholar
  12. Heine H, Lien E (2003) Toll-like receptors and their function in innate and adaptive immunity. Int Arch Allergy Immunol 130:180–192CrossRefPubMedGoogle Scholar
  13. Jackson DC, Fitzmaurice C, Sheppard RC, McMurray J, Brown LE (1995) Antigenic and immunogenic properties of synthetic peptide-based T-cell determinant polymers. Biomed Pept Proteins Nucleic Acids 1:171–176PubMedGoogle Scholar
  14. Jackson DC, Lau YF, Le T, Suhrbier A, Deliyannis G, Cheers C, Smith C, Zeng W, Brown LE (2004) A totally synthetic vaccine of generic structure that targets Toll-like receptor 2 on dendritic cells and promotes antibody or cytotoxic T cell responses. Proc Natl Acad Sci USA 101:15440–15445CrossRefPubMedGoogle Scholar
  15. Jin MS, Lee JO (2008a) Structures of the toll-like receptor family and its ligand complexes. Immunity 29:182–191CrossRefPubMedGoogle Scholar
  16. Jin MS, Lee JO (2008b) Structures of TLR-ligand complexes. Curr Opin Immunol 20:414–419CrossRefPubMedGoogle Scholar
  17. Jin MS, Kim SE, Heo JY, Lee ME, Kim HM, Paik SG, Lee H, Lee JO (2007) Crystal structure of the TLR1-TLR2 heterodimer induced by binding of a tri-acylated lipopeptide. Cell 130:1071–1082CrossRefPubMedGoogle Scholar
  18. Jung G, Bessler WG (1995) Lipopeptides as adjuvant and intergratd adjuvanticity. In: Zegers ND, Boersma WJA, Claassen E (eds) Immunological recognition of peptides in medicine and biology. CRC Press, Boca Raton, pp 159–168Google Scholar
  19. Kumagai Y, Takeuchi O, Akira S (2008) Pathogen recognition by innate receptors. J Infect Chemother 14:86–92CrossRefPubMedGoogle Scholar
  20. Lau YF, Deliyannis G, Zeng W, Mansell A, Jackson DC, Brown LE (2006) Lipid-containing mimetics of natural triggers of innate immunity as CTL-inducing influenza vaccines. Int Immunol 18:1801–1813CrossRefPubMedGoogle Scholar
  21. Lehner MD, Morath S, Michelsen KS, Schumann RR, Hartung T (2001) Induction of cross-tolerance by lipopolysaccharide and highly purified lipoteichoic acid via different Toll-like receptors independent of paracrine mediators. J Immunol 166:5161–5167PubMedGoogle Scholar
  22. Metzger J, Jung G, Bessler WG, Hoffmann P, Strecker M, Lieberknecht A, Schmidt U (1991) Lipopeptides containing 2-(palmitoylamino)-6, 7-bis(palmitoyloxy) heptanoic acid: synthesis, stereospecific stimulation of B-lymphocytes and macrophages, and adjuvanticity in vivo and in vitro. J Med Chem 34:1969–1974CrossRefPubMedGoogle Scholar
  23. Metzger JW, Beck-Sickinger AG, Loleit M, Eckert M, Bessler WG, Jung G (1995) Synthetic S-(2, 3-dihydroxypropyl)-cysteinyl peptides derived from the N-terminus of the cytochrome subunit of the photoreaction centre of Rhodopseudomonas viridis enhance murine splenocyte proliferation. J Pept Sci 1:184–190CrossRefPubMedGoogle Scholar
  24. Muhlradt PF, Kiess M, Meyer H, Sussmuth R, Jung G (1997) Isolation, structure elucidation, and synthesis of a macrophage stimulatory lipopeptide from Mycoplasma fermentans acting at picomolar concentration. J Exp Med 185:1951–1958CrossRefPubMedGoogle Scholar
  25. Muhlradt PF, Kiess M, Meyer H, Sussmuth R, Jung G (1998) Structure and specific activity of macrophage-stimulating lipopeptides from Mycoplasma hyorhinis. Infect Immun 66:4804–4810PubMedGoogle Scholar
  26. Sandor F, Latz E, Re F, Mandell L, Repik G, Golenbock DT, Espevik T, Kurt-Jones EA, Finberg RW (2003) Importance of extra- and intracellular domains of TLR1 and TLR2 in NFkappa B signaling. J Cell Biol 162:1099–1110CrossRefPubMedGoogle Scholar
  27. Schromm AB, Howe J, Ulmer AJ, Wiesmuller KH, Seyberth T, Jung G, Rossle M, Koch MH, Gutsmann T, Brandenburg K (2007) Physicochemical and biological analysis of synthetic bacterial lipopeptides: validity of the concept of endotoxic conformation. J Biol Chem 282:11030–11037CrossRefPubMedGoogle Scholar
  28. Schwandner R, Dziarski R, Wesche H, Rothe M, Kirschning CJ (1999) Peptidoglycan- and lipoteichoic acid-induced cell activation is mediated by toll-like receptor 2. J Biol Chem 274:17406–17409CrossRefPubMedGoogle Scholar
  29. Seyberth T, Voss S, Brock R, Wiesmuller KH, Jung G (2006) Lipolanthionine peptides act as inhibitors of TLR2-mediated IL-8 secretion. Synthesis and structure-activity relationships. J Med Chem 49:1754–1765CrossRefPubMedGoogle Scholar
  30. Spohn R, Buwitt-Beckmann U, Brock R, Jung G, Ulmer AJ, Wiesmuller KH (2004) Synthetic lipopeptide adjuvants and Toll-like receptor 2—structure–activity relationships. Vaccine 22:2494–2499CrossRefPubMedGoogle Scholar
  31. Takeda K, Takeuchi O, Akira S (2002) Recognition of lipopeptides by Toll-like receptors. J Endotoxin Res 8:459–463PubMedGoogle Scholar
  32. Takeuchi O, Kaufmann A, Grote K, Kawai T, Hoshino K, Morr M, Muhlradt PF, Akira S (2000) Cutting edge: preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-2 activates immune cells through a toll-like receptor 2- and MyD88-dependent signaling pathway. J Immunol 164:554–557PubMedGoogle Scholar
  33. Vitiello A, Ishioka G, Grey HM, Rose R, Farness P, LaFond R, Yuan L, Chisari FV, Furze J, Bartholomeuz R et al (1995) Development of a lipopeptide-based therapeutic vaccine to treat chronic HBV infection. I. Induction of a primary cytotoxic T lymphocyte response in humans. J Clin Invest 95:341–349CrossRefPubMedGoogle Scholar
  34. Winzler C, Rovere P, Rescigno M, Granucci F, Penna G, Adorini L, Zimmermann VS, Davoust J, Ricciardi-Castagnoli P (1997) Maturation stages of mouse dendritic cells in growth factor-dependent long-term cultures. J Exp Med 185:317–328CrossRefPubMedGoogle Scholar
  35. Zeng W, Ghosh S, Lau YF, Brown LE, Jackson DC (2002) Highly immunogenic and totally synthetic lipopeptides as self-adjuvanting immunocontraceptive vaccines. J Immunol 169:4905–4912PubMedGoogle Scholar
  36. Zeng W, Gauci S, Ghosh S, Walker J, Jackson DC (2005) Characterisation of the antibody response to a totally synthetic immunocontraceptive peptide vaccine based on LHRH. Vaccine 23:4427–4435CrossRefPubMedGoogle Scholar
  37. Zeng W, Pagnon J, Jackson DC (2007) The C-terminal pentapeptide of LHRH is a dominant B cell epitope with antigenic and biological function. Mol Immunol 44:3724–3731CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Department of Microbiology and ImmunologyThe University of MelbourneParkvilleAustralia

Personalised recommendations