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

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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.

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Acknowledgments

This study was supported by grants from the Medical Scientific Research Foundation of Guangdong Province (A2009497), and the Medical Science and Technology Program of Guangzhou (2008-YB-012, 2009-YB-021, 2009-ZDi-02, and 2008-ZDi-14).

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Authors and Affiliations

  1. Department of Anesthesiology, Guangzhou First Municipal People’s Hospital, Guangzhou Medical College, Guangzhou, 510180, China

    Ning Ding, Lixin Xu & Shouzhang She

  2. Department of Out-Patient, Guangzhou First Municipal People’s Hospital, Guangzhou Medical College, Guangzhou, 510180, China

    Hui Xiao

  3. Department of Medicine, Shandong Binzhou Vocational College, Binzhou, 256603, China

    Fang Wang

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  1. Ning Ding
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  2. Hui Xiao
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  3. Fang Wang
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  4. Lixin Xu
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  5. Shouzhang She
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Corresponding author

Correspondence to Ning Ding.

Additional information

Responsible Editor: J. Skotnicki.

N. Ding and H. Xiao contributed equally to this study.

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Ding, N., Xiao, H., Wang, F. et al. A FQHPSFI peptide selectively binds to LPS-activated alveolar macrophages and inhibits LPS-induced MIP-2 production. Inflamm. Res. 59, 627–634 (2010). https://doi.org/10.1007/s00011-010-0175-7

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  • DOI: https://doi.org/10.1007/s00011-010-0175-7

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