April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Identification of the Chemotactic Domain of the Multifunctional Inflammatory Protein CAP37, for Human Corneal Epithelial Cells
Author Affiliations & Notes
  • H. A. Pereira
    Pathology, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, Oklahoma
  • S. Logan
    Pathology, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, Oklahoma
  • H. Hinsley
    Pathology, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, Oklahoma
  • I. Hoover
    Pathology, Univ of Oklahoma Hlth Sci Ctr, Oklahoma City, Oklahoma
  • P. Svoboda
    Biotechnology Core Facility Branch, NCPDCID/CCID/CDC, Atlanta, Georgia
  • J. Pohl
    Biotechnology Core Facility Branch, NCPDCID/CCID/CDC, Atlanta, Georgia
  • Footnotes
    Commercial Relationships  H.A. Pereira, None; S. Logan, None; H. Hinsley, None; I. Hoover, None; P. Svoboda, None; J. Pohl, None.
  • Footnotes
    Support  NIH RO1 EY015534
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 5527. doi:
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      H. A. Pereira, S. Logan, H. Hinsley, I. Hoover, P. Svoboda, J. Pohl; Identification of the Chemotactic Domain of the Multifunctional Inflammatory Protein CAP37, for Human Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):5527.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : CAP37 is an inflammatory mediator that is constitutively expressed in neutrophils and plays a critical role in the innate defenses of the host. Prior studies from our laboratory have demonstrated that CAP37 has potent regulatory effects on the host inflammatory response including monocyte migration and activation. More recently we have shown that it is an effective regulator of human corneal epithelial cell (HCEC) functions including migration and proliferation. In addition to its constitutive expression in neutrophils, CAP37 is induced in corneal epithelial cells in response to infection. We hypothesize that CAP37 released by neutrophils or induced in the avascular cornea plays a role in the migration of monocytes from the circulation to the cornea and can also promote corneal epithelial cell migration thus promoting corneal healing in response to wounding or infection. The purpose of this study was to delineate the chemotactic domain of CAP37.

Methods: : Overlapping peptides based on the complete amino acid sequence of CAP37 were synthesized. Each peptide was tested for chemotactic activity for HCEC, human peripheral blood monocytes, neutrophils, and lymphocytes and the mouse macrophage cell line RAW264.7 using the modified Boyden chamber chemotaxis assay.

Results: : Our results demonstrated that a peptide consisting of amino acids 95 through 122 of the native protein had strong chemotactic activity for HCEC, human monocytes, and mouse macrophages, but had no effect on neutrophil and lymphocyte migration. Truncations and extensions of the peptide indicated that the critical residues required for chemotaxis lie between amino acids 102-122. Dose response studies indicated that maximal chemotactic activity was obtained at peptide concentrations between 10-4 and 10-6 M. A monoclonal antibody to CAP37 attenuated the chemotactic activity of the peptides.

Conclusions: : We have identified the chemotactic domain of the multifunctional inflammatory protein CAP37 and shown it to be distinct from the previously described bactericidal domain of the molecule.

Keywords: inflammation • cornea: epithelium • protein structure/function 
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