May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Lipocalin-1 Associates With Lysozyme and With Endogenous Antibodies in Tears
Author Affiliations & Notes
  • S. G. Remington
    Ophthalmology Research, Regions Hospital, St Paul, Minnesota
  • J. M. Crow
    Ophthalmology Research, Regions Hospital, St Paul, Minnesota
  • J. D. Nelson
    Ophthalmology Research, Regions Hospital, St Paul, Minnesota
  • Footnotes
    Commercial Relationships S.G. Remington, None; J.M. Crow, None; J.D. Nelson, None.
  • Footnotes
    Support HealthPartners Research Foundation
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 1907. doi:https://doi.org/
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      S. G. Remington, J. M. Crow, J. D. Nelson; Lipocalin-1 Associates With Lysozyme and With Endogenous Antibodies in Tears. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1907. doi: https://doi.org/.

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

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Abstract

Purpose:: To address the association of lipocalin-1 with other proteins in ocular tears.

Methods:: 1) Western blots: Human tear samples were separated by polyacrylamide gel electrophoresis (PAGE) in 15% gels under reducing and non-reducing conditions, and transferred to nitrocellulose. The blots were sequentially incubated with anti-human lipocalin-1 primary antibodies (R&D Systems), alkaline phosphatase-conjugated secondary antibodies (Sigma), and NBT/BCIP.2) Immuno-selection: Human tears were incubated with anti-human lipocalin-1 antibodies and Protein G sepharose (Amersham Biosciences). The immune complex proteins were eluted, separated by PAGE and silver stained.

Results:: 1) In Western blots, antibodies to lipocalin-1 recognized reduced lipocalin-1 monomer (17 and 18kD bands), and non-reduced lipocalin-1 dimer (37kD). In heated, non-reduced tear samples, antibodies to lipocalin-1 also recognized other higher molecular weight protein complexes.2) Immuno-selection of lipocalin-1 resulted in co-selection of lysozyme. Controls with no added antibodies also resulted in immuno-selection of lipocalin-1 and lysozyme, implying that normal tears contain antibodies to lipocalin-1 and/or lysozyme.

Conclusions:: Lipocalin-1 associates with lysozyme and other proteins in tears. Normal tears contain antibodies to lipocalin-1. Auto-antibodies in tears may be an attribute of normal ocular homeostasis.

Keywords: cornea: tears/tear film/dry eye • protein purification and characterization 
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