Inflammation Research

, Volume 59, Issue 8, pp 627–634

A FQHPSFI peptide selectively binds to LPS-activated alveolar macrophages and inhibits LPS-induced MIP-2 production

  • Ning Ding
  • Hui Xiao
  • Fang Wang
  • Lixin Xu
  • Shouzhang She
Original Research Paper



The goal of this study was to identify peptides selectively binding to lipopolysaccharide (LPS)-activated alveolar macrophages (AMs) and to characterize their effects on the production of LPS-induced cytokines.


A phage display library was sequentially screened by binding phages to unmanipulated AMs and then to LPS-activated AMs. Individual phage clones were identified by cell-based ELISA. Positive phage clones were characterized by DNA sequencing and bioinformatics analysis. Binding specificity of the selected phage to LPS-activated AMs was tested using immunofluorescent staining. The selected candidate peptide was chemically synthesized to determine whether it could modulate LPS-induced cytokine production in AMs.


Twenty-two out of 40 phage clones selected randomly after four rounds of biopanning bound selectively to LPS-activated AMs, and 12 of them displayed novel peptides. A phage clone displaying FQHPSFI peptide bound effectively to LPS-activated AMs, but not to other cells tested. Furthermore, the synthetic FQHPSFI peptide, but not seven point mutants tested, competitively inhibited the binding of the phage clone to LPS-activated AMs. Importantly, the FQHPSFI peptide significantly inhibited LPS-stimulated microphage inflammatory protein 2 (MIP-2) production in vitro.


Our data demonstrate that phage display technology is a powerful tool for the identification of bioactive peptides. The identified FQHPSFI peptide may be used for the modulation of LPS-stimulated MIP-2 production in AMs.


Alveolar macrophages Peptide Phage display Lipopolysaccharide Inflammation 


  1. 1.
    Gao H, Evans TW, Finney SJ. Bench-to-bedside review: sepsis, severe sepsis and septic shock—does the nature of the infecting organism matter? Crit Care. 2008;12:213.CrossRefPubMedGoogle Scholar
  2. 2.
    Rubenfeld GD, Caldwell E, Peabody E, Weaver J, Martin DP, Neff M, et al. Incidence and outcomes of acute lung injury. N Engl J Med. 2005;53:1685–93.CrossRefGoogle Scholar
  3. 3.
    Lin WJ, Yeh WC. Implication of Toll-like receptor and tumor necrosis factor alpha signaling in septic shock. Shock. 2005;24:206–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Dong L, Wang S, Chen M, Li H, Bi W. The activation of macrophage and upregulation of CD40 costimulatory molecule in lipopolysaccharide-induced acute lung injury. J Biomed Biotechnol. 2008;2008:852571.PubMedGoogle Scholar
  5. 5.
    Puneet P, Moochhala S, Bhatia M. Chemokines in acute respiratory distress syndrome. Am J Physiol Lung Cell Mol Physiol. 2005;288:L3–15.CrossRefPubMedGoogle Scholar
  6. 6.
    Engwegen JY, Gast MC, Schellens JH, Beijnen JH. Clinical proteomics: searching for better tumour markers with SELDI-TOF mass spectrometry. Trends Pharmacol Sci. 2006;27:251–9.CrossRefPubMedGoogle Scholar
  7. 7.
    Kolonin MG, Pasqualini R, Arap W. Molecular addresses in blood vessels as targets for therapy. Curr Opin Chem Biol. 2001;5:308–13.CrossRefPubMedGoogle Scholar
  8. 8.
    Arap W, Pasqualini R, Ruoslahti E. Cancer treatment by targeted drug delivery to tumor vasculature in a mouse model. Science. 1998;279:377–80.CrossRefPubMedGoogle Scholar
  9. 9.
    Gatto B, Cavalli M. From proteins to nucleic acid-based drugs: the role of biotech in anti-VEGF therapy. Anticancer Agents Med Chem. 2006;6:287–301.CrossRefPubMedGoogle Scholar
  10. 10.
    Dos Santos S, Delattre AI, De Longueville F, Bult H, Raes M. Gene expression profiling of LPS-stimulated murine macrophages and role of the NF-kappaB and PI3K/mTOR signaling pathways. Ann N Y Acad Sci. 2007;1096:70–7.CrossRefPubMedGoogle Scholar
  11. 11.
    Huang JH, Lu L, Lu H, Chen X, Jiang S, Chen YH. Identification of the HIV-1 gp41 core-binding motif in the scaffolding domain of caveolin-1. J Biol Chem. 2007;282:6143–52.CrossRefPubMedGoogle Scholar
  12. 12.
    Li Z, Nardi MA, Karpatkin S. Role of molecular mimicry to HIV-1 peptides in HIV-1 related immunologic thrombocytopenia. Blood. 2005;106:572–6.CrossRefPubMedGoogle Scholar
  13. 13.
    Bishop-Hurley SL, Schmidt FJ, Erwin AL, Smith AL. Peptides selected for binding to a virulent strain of haemophilus influenzae by phage display are bactericidal. Antimicrob Agents Chemother. 2005;49:2972–8.CrossRefPubMedGoogle Scholar
  14. 14.
    Stamme C, Walsh E, Wright JR. Surfactant protein A differentially regulates IFN-gamma- and LPS-induced nitrite production by rat alveolar macrophages. Am J Respir Cell Mol Biol. 2000;23:772–9.PubMedGoogle Scholar
  15. 15.
    Giordano RJ, Cardo-Vila M, Lahdenranta J, Pasqualini R, Arap W. Biopanning and rapid analysis of selective interactive ligands. Nat Med. 2001;7:1249–53.CrossRefPubMedGoogle Scholar
  16. 16.
    Zhang Y, Chen J, Zhang Y, Hu Z, Hu D, Pan Y, et al. Panning and identification of a colon tumor binding peptide from a phage display peptide library. J Biomol Screen. 2007;12:429–35.CrossRefPubMedGoogle Scholar
  17. 17.
    de Jager W, te Velthuis H, Prakken BJ, Kuis W, Rijkers GT. Simultaneous detection of 15 human cytokines in a single sample of stimulated peripheral blood mononuclear cells. Clin Diagn Lab Immunol. 2003;10:133–9.PubMedGoogle Scholar
  18. 18.
    Rensen PC, Gras JC, Lindfors EK, van Dijk KW, Jukema JW, van Berkel TJ, et al. Selective targeting of liposomes to macrophages using a ligand with high affinity for the macrophage scavenger receptor class A. Curr Drug Discov Technol. 2006;3:135–44.CrossRefPubMedGoogle Scholar
  19. 19.
    Nowakowski GS, Dooner MS, Valinski HM, Mihaliak AM, Quesenberry PJ, Becker PS. A specific heptapeptide from a phage display peptide library homes to bone marrow and binds to primitive hematopoietic stem cells. Stem Cells. 2004;22:1030–8.CrossRefPubMedGoogle Scholar
  20. 20.
    Triantafilou M, Brandenburg K, Kusumoto S, Fukase K, Mackie A, Seydel U, et al. Combinational clustering of receptors following stimulation by bacterial products determines LPS responses. Biochem J. 2004;381:527–36.CrossRefPubMedGoogle Scholar
  21. 21.
    Hartley O. The use of phage display in the study of receptors and their ligands. J Recept Signal Transduct Res. 2002;22:373–92.CrossRefPubMedGoogle Scholar
  22. 22.
    Jost PJ, Harbottle RP, Knight A, Miller AD, Coutelle C, Schneider H. A novel peptide, THALWHT, for the targeting of human airway epithelia. FEBS Lett. 2001;489:263–9.CrossRefPubMedGoogle Scholar
  23. 23.
    Eda K, Eda S, Sherman IW. Identification of peptides targeting the surface of plasmodium falciparum-infected erythrocytes using a phage display peptide library. Am J Trop Med Hyg. 2004;71:190–5.PubMedGoogle Scholar

Copyright information

© Springer Basel AG 2010

Authors and Affiliations

  • Ning Ding
    • 1
  • Hui Xiao
    • 2
  • Fang Wang
    • 3
  • Lixin Xu
    • 1
  • Shouzhang She
    • 1
  1. 1.Department of AnesthesiologyGuangzhou First Municipal People’s Hospital, Guangzhou Medical CollegeGuangzhouChina
  2. 2.Department of Out-PatientGuangzhou First Municipal People’s Hospital, Guangzhou Medical CollegeGuangzhouChina
  3. 3.Department of MedicineShandong Binzhou Vocational CollegeBinzhouChina

Personalised recommendations