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
  • 97 Downloads

Abstract

Objective

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.

Methods

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.

Results

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.

Conclusions

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.

Keywords

Alveolar macrophages Peptide Phage display Lipopolysaccharide Inflammation 

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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

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