E-cadherin distribution and epithelial basement membrane characteristics of the normal human conjunctiva and cornea.

The conjunctival mucosa has several similarities to the mucosal immune system of the gut and bronchus. Like the gut and bronchial mucosa, the conjunctiva is capable of inducing tolerance to encountered antigens and possesses a repertoire of CD8+ intraepithelial lymphocytes (IELs) bearing the human mucosal lymphocyte-1 antigen (HML-1) which has been shown to be an alpha E beta 7 integrin. The epithelial cells surface ligand for HML-1 is E-cadherin. The distribution of E-cadherin in the normal human conjunctiva and cornea is not known. We investigated E-cadherin distribution in the conjunctiva and cornea by immunohistochemistry. E-cadherin was found to be present in all layers of the conjunctival epithelium but not in corneal epithelium. In the conjunctiva it may act as a ligand for the HML-1+ IELs. The specific location of IELs along the basal cells of the conjunctiva compared with the generalised distribution of E-cadherin through all layers, indicates that factors other than E-cadherin binding determine the distribution of HML-1+ IELs. We performed electron microscopy on de-epithelialised conjunctival and corneal samples. We demonstrated the presence of epithelial basement membrane pores in the conjunctiva but not in the cornea. Lymphocyte migration from the substantia propria to the intraepithelial compartment appears to occur through these pores, which may also serve as a conduit for antigen presentation by epithelial antigen presenting cells (APCs) to lymphocytes in the substantia propria.

A new epithelial cell type in the human cornea.

PURPOSE: To study the expression of intermediate filaments in the human cornea. METHODS: Light and electron microscopic and immunohistochemical studies were performed on 20 corneas from subjects of various ages. RESULTS: A hitherto unrecognized epithelial cell population emerged from the immunohistochemical studies. Epithelial cells were invariably present in the superior cornea, whereas the nasal, temporal, and inferior segments almost lacked these cells. They were situated at the transition between peripheral cornea and limbus, and occurred as small groups in the basal epithelium. On electron microscopy, they were recognized by their marginated nuclear chromatin, large nucleoli, prominent bundles of intermediate filaments, and numerous hemidesmosomes and desmosomes. Immunohistochemistry on frozen sections revealed a unique intermediate filament make-up: ie, strong co-expression of vimentin and cytokeratin 19; other intermediate filaments, including cytokeratins 3, 4, 6, 7, 8, 10, 13, and 18 were negative. Finally, the cells lacked ultrastructural and immunohistochemical features of melanocytes, neuroendocrine cells, Langerhans' cells, and leukocytes. CONCLUSIONS: A new epithelial cell type in the human cornea is described with characteristic morphologic and immunohistochemical features. According to their particular segmental distribution, restricted localization at the junction between cornea and limbus, and expression of an "early" intermediate filament profile, it is tempting to speculate that they represent stem cells of the human cornea. Further studies are aimed to characterize their phenotype and function more extensively.

The transitional zone between limbus and peripheral cornea. An immunohistochemical study.

PURPOSE: The authors investigated the phenotypic characteristics of basally located "transitional cells" in peripheral superior cornea, characterized previously by their coexpression of cytokeratin 19 and vimentin and their negativity for AE5. METHODS: Twenty adult human corneas were studied, using in situ immunohistochemical techniques and a panel of specific monoclonal antibodies against various surface and cytoplasmic molecules. RESULTS: The transitional cells shared staining characteristics with limbal basal cells in their expression of alpha 6 beta 4-integrin, metallothionein, AE1, and transferrin receptor. CONCLUSIONS: These "transitional" epithelial cells exhibit a unique phenotype differing from that of the surrounding basal epithelial cells in the peripheral cornea but analogous to that of limbal basal cells. These findings further corroborate the hypothesis that, at least from an immunohistochemical point of view, transitional cells in the superior peripheral cornea exhibit stem cell characteristics.

Localization of the cytoadhesin integrins in the human cornea. An immunohistochemical study using monoclonal antibodies.

Cell-matrix interactions play a fundamental role in normal and pathological conditions. They can be mediated by the cytoadhesin subgroup of the integrin superfamily of adhesion molecules. Its members include the vitronectin receptor (VNR) and the platelet glycoprotein IIb/IIIa (GP IIb/IIIa). Both receptors are composed of an alpha-chain (alpha v and alpha IIb, respectively) coupled to a beta 3-chain. Using in situ immunohistochemistry and monoclonal antibodies, the authors studied the distribution of GP IIIa (common beta 3-chain), GP IIb/IIIa (alpha IIb-chain) and VNR (alpha v-chain) in normal and pathological corneal tissues. In the normal cornea, the limbal vascular endothelium was weakly alpha v-positive. Occasionally, faint and granular staining was seen in the epithelium. In the pathological samples, an upregulated expression of the alpha v-chain was noticed on the corneal epithelium as well as on fibroblasts and corneal endothelium. The alpha IIb and beta 3-chains were consistently absent. These data suggest that expression of the VNR-alpha v-chain in the human cornea is modulated by soluble factors released during inflammation and wound healing. Dissociation of expression of the alpha v and beta 3-chains suggests usage of an alternative beta-chain by the VNR-alpha v-chain.

Distribution of very late activation integrins in the human cornea. An immunohistochemical study using monoclonal antibodies.

There is growing evidence that cellular adhesion mechanisms characterized by cell-cell and cell-matrix interactions are a fundamental process in the immunobiology of the cornea. Interactions with various extracellular matrix components are mediated by the very late activation (VLA) subgroup of the integrin superfamily of adhesion molecules. The six different VLA dimers known thus far consist of a common beta 1 subunit and a variable alpha (1 to 6) subunit. They serve as receptors for laminin (alpha 3 and alpha 6), collagen (alpha 2 and alpha 3), and fibronectin (alpha 4 and alpha 5). Using in situ immunohistochemistry and monoclonal antibodies, the distribution of the common beta 1 and the variable alpha-chains of VLA molecules was studied in normal human cornea and in cases with scarring or subepithelial/retrocorneal fibrous tissue. Epithelial cells were VLA-beta 1 and VLA-alpha 2, -alpha 3, -alpha 4, -alpha 5, and -alpha 6 positive. This is consistent with their intercellular adhesion and may aid in their attachment to the basement membrane which is composed of collagen, laminin, and fibronectin. Keratocytes in normal stroma expressed only the common beta 1-chain and no detectable alpha-chains. In regions of scar or fibrous tissue, however, an upregulated expression of the alpha-chains was detected. The VLA- alpha 1, -alpha 3, -alpha 4, and -alpha 5 were expressed in young fibrous tissue; in older lesions, VLA- alpha 1, -alpha 2, -alpha 3, -alpha 4, and -alpha 5 could be detected. The corneal endothelium showed a strikingly strong positivity for all VLA integrins.(ABSTRACT TRUNCATED AT 250 WORDS)