Abstract
In response to an unexpected observation of apparent localisation by immunocytochemistry, we have investigated the potential expression and function of P-selectin (CD62P) in human corneal epithelial cells. The SV40 immortalised cell line, HCE-T (validated by STR profiling), along with multiple donor corneal-limbal tissue samples, were examined for P-selectin expression using a combination of immunocytochemistry, Western blotting, RT-PCR and immunohistochemistry. Potential expression of the major ligand for P-selectin (P-selectin glycoprotein ligand-1; PSGL-1; CD162) was also examined by immunocytochemistry and RT-PCR. A selective inhibitor of P-selectin-PSGL-1 binding (KF38789) was subsequently tested for effects on HCE-T cells using a cell culture gap-closure assay. HCE-T cells as well as primary epithelial cultures derived from donor corneal-limbal tissue, displayed positive immunostaining for P-selectin. Staining was particularly evident at cell-cell boundaries and at the outer edge of expanding epithelial islands. P-selectin expression was confirmed by Western blotting and RT-PCR (validated by product sequencing), as well as by immunohistochemistry performed on serial sections of corneal-limbal tissue stained for P-selectin...Continue Reading
References
Sep 1, 1985·The Journal of Cell Biology·P E StenbergD F Bainton
Jul 1, 1986·The Journal of Cell Biology·A SchermerT T Sun
Jul 1, 1989·The Journal of Clinical Investigation·R P McEverD F Bainton
Oct 20, 1989·Cell·E LarsenB Furie
Jan 1, 1994·Ophthalmology·M F GoldbergJ S Pepose
Apr 27, 1999·Progress in Retinal and Eye Research·M E Fini
Jun 1, 2000·Cornea·S E Wilson
Jan 25, 2003·The British Journal of Ophthalmology·M IwataM Sawa
Jun 28, 2003·Trends in Molecular Medicine·Klaus Ley
Nov 24, 2004·Investigative Ophthalmology & Visual Science·H Anne PereiraJames Chodosh
Nov 24, 2005·Investigative Ophthalmology & Visual Science·Satoru YamagamiNobuyuki Ebihara
Oct 27, 2006·Investigative Ophthalmology & Visual Science·Zhijie LiC Wayne Smith
Jul 1, 2007·Cell Adhesion & Migration·Tao XuJian-Guo Geng
Jul 18, 2009·The American Journal of Pathology·Sarah E ByesedaZhijie Li
Oct 26, 2010·Experimental Eye Research·S Louise AinscoughDamien G Harkin
Dec 21, 2010·American Journal of Physiology. Heart and Circulatory Physiology·Fong W LamRolando E Rumbaut
Sep 20, 2011·The American Journal of Pathology·Nan GaoFu-Shin X Yu
Dec 19, 2012·Experimental Eye Research·Thomas A HogerheydeNeil A Richardson
Aug 27, 2013·Developmental Cell·Mei-Ling Chin, Marek Mlodzik
Oct 8, 2013·Cytotherapy·Laura J BrayDamien G Harkin
Dec 17, 2014·Experimental Eye Research·Peter J GilliesDamien G Harkin
Mar 17, 2015·PloS One·Fong W LamAlan R Burns
Jul 23, 2015·Investigative Ophthalmology & Visual Science·Maud GorbetSara Williams
Apr 18, 2016·Experimental Eye Research·Fiona J LiDamien G Harkin
Jul 7, 2017·BioMed Research International·János Kappelmayer, Béla Nagy