A FQHPSFI peptide selectively binds to LPS-activated alveolar macrophages and inhibits LPS-induced MIP-2 production
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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.
- 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. CrossRef
- 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. CrossRef
- Lin WJ, Yeh WC. Implication of Toll-like receptor and tumor necrosis factor alpha signaling in septic shock. Shock. 2005;24:206–9. CrossRef
- 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.
- Puneet P, Moochhala S, Bhatia M. Chemokines in acute respiratory distress syndrome. Am J Physiol Lung Cell Mol Physiol. 2005;288:L3–15. CrossRef
- 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. CrossRef
- Kolonin MG, Pasqualini R, Arap W. Molecular addresses in blood vessels as targets for therapy. Curr Opin Chem Biol. 2001;5:308–13. CrossRef
- Arap W, Pasqualini R, Ruoslahti E. Cancer treatment by targeted drug delivery to tumor vasculature in a mouse model. Science. 1998;279:377–80. CrossRef
- 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. CrossRef
- 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. CrossRef
- 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. CrossRef
- 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. CrossRef
- 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. CrossRef
- 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.
- 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. CrossRef
- 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. CrossRef
- 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.
- 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. CrossRef
- 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. CrossRef
- 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. CrossRef
- Hartley O. The use of phage display in the study of receptors and their ligands. J Recept Signal Transduct Res. 2002;22:373–92. CrossRef
- 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. CrossRef
- 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.
- A FQHPSFI peptide selectively binds to LPS-activated alveolar macrophages and inhibits LPS-induced MIP-2 production
Volume 59, Issue 8 , pp 627-634
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- SP Birkhäuser Verlag Basel
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- Alveolar macrophages
- Phage display
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- Author Affiliations
- 1. Department of Anesthesiology, Guangzhou First Municipal People’s Hospital, Guangzhou Medical College, Guangzhou, 510180, China
- 2. Department of Out-Patient, Guangzhou First Municipal People’s Hospital, Guangzhou Medical College, Guangzhou, 510180, China
- 3. Department of Medicine, Shandong Binzhou Vocational College, Binzhou, 256603, China