Detection of galectin-3 in tear fluid at disease states and immunohistochemical and lectin histochemical analysis in human corneal and conjunctival epithelium
- Enkela Hrdličková-Celaa,b,
- Jan Plzákb,c,
- Karel Smetana, Jrb,d,
- Zora Mělkováe,
- Herbert Kaltnerf,
- Martin Filipeca,
- Fu-Tong Liug,
- Hans-Joachim Gabiusf
- aCharles University, 1st Faculty of Medicine, Department of Ophthalmology, Prague, Czech Republic, b1st Faculty of Medicine, Institute of Anatomy, c1st Faculty of Medicine, Department of Otorhino- laryngology, Head and Neck Surgery, dCentre for Cell Therapy and Tissue Repair, e1st Faculty of Medicine, Department of Pathological Physiology, fLudwig-Maximilians- University, Faculty of Veterinary Medicine, Institute of Physiological Chemistry, Munich, Germany, gLa Jolla Institute for Allergy and Immunology, San Diego, CA, USA
- Karel Smetana, Charles University, 1st Faculty of Medicine, Institute of Anatomy, U nemocnice 3, 128 00 Prague 2, Czech Republic ksmet{at}lf1.cuni.cz
- Accepted 16 May 2001
Abstract
BACKGROUND/AIM Components of the tear fluid contribute to the biochemical defence system of the eye. To reveal whether the immune mediator and lipopolysaccharide binding galectin-3 is present in tears, tear samples were collected from eyes in healthy and pathological states. Investigation of expression of galectin-3 and galectin-3 reactive glycoligands in normal human conjunctival and corneal epithelia was also initiated as a step to understand the role of galectin-3 in ocular surface pathology.
METHODS Immunoblot analysis using either a rabbit polyclonal or a mouse monoclonal antibody against galectin-3 was employed to detect galectin-3 in tear fluid. Galectin-3 expression in tissue specimens was detected by immunocytochemistry employing A1D6 mouse monoclonal antibody, and galectin-3 reactive glycoligands were visualised by lectin histochemistry using labelled galectin-3.
RESULTS Galectin-3 was found only in tears from patients with ocular surface disorders. It was expressed in normal corneal and conjunctival epithelia but not in lacrimal glands. Inflammatory leucocytes and goblet cells found in galectin-3 containing tear fluid also expressed galectin-3. Galectin-3 binding sites were detected on the surface of conjunctival and corneal epithelial cells co-localising with desmoglein.
CONCLUSIONS This study revealed expression of galectin-3 in tear fluid obtained from patients with eye diseases. The role of this endogenous lectin (produced by inflammatory as well as epithelial cells) in antimicrobial action and inflammation modulation could be expected.
