The lectin KM+ induces corneal epithelial wound healing in rabbits

Neutrophil influx is essential for corneal regeneration ( Gan et al. 1999 ). KM+, a lectin from Artocarpus integrifolia, induces neutrophil migration ( Santos‐de‐Oliveira et al. 1994 ). This study aims at investigating a possible effect of KM+ on corneal regeneration in rabbits. A 6.0‐mm diameter area of debridement was created on the cornea of both eyes by mechanical scraping. The experimental eyes received drops of KM+ (2.5 μg/ml) every 2 h. The control eyes received buffer. The epithelial wounded areas of the lectin‐treated and untreated eyes were stained with fluorescein, photographed and measured. The animals were killed 12 h (group 1, n = 5), 24 h (group 2, n = 10) and 48 h (group 3, n = 5) after the scraping. The corneas were analysed histologically (haematoxylin and eosin and immunostaining for proliferation cell nuclear antigen, p63, vascular endothelial growth factor, c‐Met and laminin). No significant differences were found at the epithelial gap between treated and control eyes in the group 1. However, the number of neutrophils in the wounded area was significantly higher in treated eyes in this group. Three control and seven treated eyes were healed completely and only rare neutrophils persisted in the corneal stroma in group 2. No morphological distinction was observed between treated and control eyes in group 3. In treated corneas of group 2, there was an increase in immunostaining of factors involved in corneal healing compared to controls. Thus, topical application of KM+ may facilitate corneal epithelial wound healing in rabbits by means of a mechanism that involves increased influx of neutrophils into the wounded area induced by the lectin.

[1]  K. Resing,et al.  KM+, a mannose‐binding lectin from artocarpus integrifolia: Amino acid sequence, predicted tertiary structure, carbohydrate recognition, and analysis of the β‐prism fold , 2008, Protein science : a publication of the Protein Society.

[2]  L. O’Neill,et al.  Toll‐like receptors: key activators of leucocytes and regulator of haematopoiesis , 2007, British journal of haematology.

[3]  D. Wakefield,et al.  A critical role for CCL2 and CCL3 chemokines in the regulation of polymorphonuclear neutrophils recruitment during corneal infection in mice , 2007, Immunology and cell biology.

[4]  E. Fuchs,et al.  p63: revving up epithelial stem-cell potential , 2007, Nature Cell Biology.

[5]  J. Nadel,et al.  Regulation of interleukin-8 via an airway epithelial signaling cascade. , 2007, American journal of physiology. Lung cellular and molecular physiology.

[6]  C. Smith,et al.  Two waves of neutrophil emigration in response to corneal epithelial abrasion: distinct adhesion molecule requirements. , 2006, Investigative ophthalmology & visual science.

[7]  F. Drago,et al.  Effects of COOH–Terminal Tripeptide –MSH11–13 on Corneal Epithelial Wound Healing: Role of Nitric Oxide , 2006 .

[8]  K. Wilhelmus,et al.  Inflammatory response to fungal keratitis. , 2005, The ocular surface.

[9]  S. Dower,et al.  Regulation of human neutrophil chemokine receptor expression and function by activation of Toll‐like receptors 2 and 4 , 2005, Immunology.

[10]  R. Mortara,et al.  Lectin KM+-induced neutrophil haptotaxis involves binding to laminin. , 2005, Biochimica et biophysica acta.

[11]  T. Welte,et al.  Life after corpse engulfment: phagocytosis of apoptotic cells leads to VEGF secretion and cell growth , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[12]  H. Sheardown,et al.  Growth factors in the anterior segment: role in tissue maintenance, wound healing and ocular pathology. , 2004, Experimental eye research.

[13]  J. Palmblad,et al.  Vascular endothelial growth factor (VEGF) and its receptor VEGFR-2 in the regulation of corneal neovascularization and wound healing. , 2004, Acta ophthalmologica Scandinavica.

[14]  J. Zieske,et al.  Cell cycle regulators at the ocular surface. , 2004, Experimental eye research.

[15]  Renato Ambrósio,et al.  The Corneal Wound Healing Response: Cytokine-mediated Interaction of the Epithelium, Stroma, and Inflammatory Cells , 2001, Progress in Retinal and Eye Research.

[16]  F. Bazzoni,et al.  The neutrophil as a cellular source of chemokines , 2000, Immunological reviews.

[17]  M. Macsai.  The Management of Corneal Trauma: Advances in the Past Twenty-five Years , 2000, Cornea.

[18]  C. Humpel,et al.  Expression of vascular endothelial growth factor and its receptors in inflamed and vascularized human corneas. , 2000, Investigative ophthalmology & visual science.

[19]  A. Haddad.  Renewal of the Rabbit Corneal Epithelium as Investigated by Autoradiography After Intravitreal Injection of 3H-thymidine , 2000, Cornea.

[20]  J. Wang,et al.  Proinflammatory mediators stimulate neutrophil-directed angiogenesis. , 1999, Archives of surgery.

[21]  H. J. Kim,et al.  Effect of leukocytes on corneal cellular proliferation and wound healing. , 1999, Investigative Ophthalmology and Visual Science.

[22]  C. Chan,et al.  Inflammatory response in the early stages of wound healing after excimer laser keratectomy. , 1998, Archives of ophthalmology.

[23]  S. Heegaard,et al.  The healing effect of all-trans retinoic acid on epithelial corneal abrasions in rabbits. , 1998, Acta ophthalmologica Scandinavica.

[24]  M. Roque-Barreira,et al.  Neutrophil haptotaxis induced by the lectin KM+ , 1998, Glycoconjugate Journal.

[25]  R. Mohan,et al.  Hepatocyte growth factor and hepatocyte growth factor receptor in the lacrimal gland, tears, and cornea. , 1996, Investigative ophthalmology & visual science.

[26]  W. Liles,et al.  Review: nomenclature and biologic significance of cytokines involved in inflammation and the host immune response. , 1995, The Journal of infectious diseases.

[27]  J. Zieske,et al.  Effect of epidermal growth factor, hepatocyte growth factor, and keratinocyte growth factor, on proliferation, motility and differentiation of human corneal epithelial cells. , 1994, Experimental eye research.

[28]  P. McDonnell,et al.  Immunochemistry with 5‐Bromo-2‐Deoxyuridine for Visualization of Mitotic Cells in the Corneal Epithelium , 1994, Cornea.

[29]  M. Roque-Barreira,et al.  A neutrophil migration-inducing lectin from Artocarpus integrifolia. , 1994, Journal of immunology.

[30]  S. Wilson,et al.  Hepatocyte growth factor, keratinocyte growth factor, their receptors, fibroblast growth factor receptor-2, and the cells of the cornea. , 1993, Investigative ophthalmology & visual science.

[31]  L. Orci,et al.  Identification of a fibroblast-derived epithelial morphogen as hepatocyte growth factor , 1991, Cell.

[32]  K. Matsumoto,et al.  Marked stimulation of growth and motility of human keratinocytes by hepatocyte growth factor. , 1991, Experimental cell research.

[33]  W. Birchmeier,et al.  Evidence for the identity of human scatter factor and human hepatocyte growth factor. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[34]  J. Rubin,et al.  A broad-spectrum human lung fibroblast-derived mitogen is a variant of hepatocyte growth factor. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[35]  D. Spriggs,et al.  Circulating human peripheral blood granulocytes synthesize and secrete tumor necrosis factor alpha. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[36]  K. Tashiro,et al.  Molecular cloning and expression of human hepatocyte growth factor , 1989, Nature.

[37]  T. Sun,et al.  Existence of slow-cycling limbal epithelial basal cells that can be preferentially stimulated to proliferate: Implications on epithelial stem cells , 1989, Cell.

[38]  C. P. Leblond,et al.  Immunoelectron microscopy of endothelial cells in rat incisor suggests that most basement membrane components are produced by young cells, whereas heparan sulfate proteoglycan is produced by both young and old cells. , 1988, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[39]  P. Morméde,et al.  Immunoreactive fibroblast growth factor in cells of peritoneal exudate suggests its identity with macrophage-derived growth factor. , 1985, Biochemical and biophysical research communications.

[40]  C. P. Leblond,et al.  Immunohistochemical evidence for the intracellular formation of basement membrane collagen (type IV) in developing tissues. , 1980, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[41]  H. Brewitt.  SLIDING OF EPITHELIUM IN EXPERIMENTAL CORNEAL WOUNDS , 1979, Acta ophthalmologica.

[42]  Oliver H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[43]  A. Hutcheon,et al.  Examination of the restoration of epithelial barrier function following superficial keratectomy. , 2007, Experimental eye research.

[44]  M. Roque-Barreira,et al.  Neutrophil activation induced by the lectin KM+ involves binding to CXCR2. , 2006, Biochimica et biophysica acta.

[45]  F. Drago,et al.  Effects of the COOH-terminal tripeptide alpha-MSH(11-13) on corneal epithelial wound healing: role of nitric oxide. , 2006, Experimental eye research.

[46]  H. Grossniklaus,et al.  Long-term histopathologic findings in human corneal wounds after refractive surgical procedures. , 2005, American journal of ophthalmology.

[47]  Alexandre Cloutier,et al.  Transcription factor activation in human neutrophils. , 2003, Chemical immunology and allergy.

[48]  H. Yee,et al.  Polymorphonuclear neutrophils promote rFGF-2-induced angiogenesis in vivo. , 2003, The Journal of surgical research.

[49]  L. Gibson,et al.  Interleukin-6 production by human neutrophils after Fc-receptor cross-linking or exposure to granulocyte colony-stimulating factor. , 1998, Blood.

[50]  G. V. van Setten.  Vascular endothelial growth factor (VEGF) in normal human corneal epithelium: detection and physiological importance. , 1997, Acta ophthalmologica Scandinavica.

[51]  G. Schultz,et al.  Growth factors and ocular wound healing , 1994, Eye.

[52]  L. Gollahon,et al.  Hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), and their receptors in human breast cells and tissues: alternative receptors. , 1994, Cellular & molecular biology research.

[53]  F. Kruse.  Stem cells and corneal epithelial regeneration , 1994, Eye.