Nicholas Dushku

Immunohistochemical evidence that human pterygia originate from an invasion of vimentin-expressing altered limbal epithelial basal cells

The goal of this study was to determine the cell origin of human pterygia. In order to determine the origin of these cells, longitudinal cryostat sections through five primary and two recurrent pterygia were studied immunohistochemically by finding limbal basal stem cell staining patterns as defined by monoclonal antibodies AE1 (staining positive) and AE5 (staining negative). In addition, sections were stained with antivimentin antibody. Altered limbal basal cells invading normal cornea along the basement membrane were identified in seven human pterygia with these specific monoclonal antibodies. A group of limbal basal cells (vimentin and AE1 positive) was always present between the dissolved edge of Bowmans layer and vascularized conjunctiva which contained goblet cells. Scattered patches of cells staining positive with both vimentin and AE5 (in addition to their AE1 staining) were also found in conjunctival epithelium growing on corneal basement membrane adjacent to the migrating limbal cells, indicating local infiltration by the altered limbal basal cells. This same pattern was also found in recurrent pterygia. Based on this data we propose that the pathogenesis of pterygia is due to a normal stationary parental limbal epithelial basal cell becoming altered and giving rise to a zone of motile daughter cells, the pterygium cells, which leave the limbal region and migrate as a group centripetally along the corneal basement membrane dissolving Bowmans layer. Since these altered limbal basal cells are found at the microscopic advancing edge over Bowmans layer with no fibroblast mass under them, the pterygium cell apparently precedes the rapid growth of the fibroblasts from the stroma.(ABSTRACT TRUNCATED AT 250 WORDS)

What a study of pterygia teaches us about the cornea? Molecular mechanisms of formation.

Experiments were carried out in the early 1990s to investigate the cell types involved in a pterygium and to determine a possible mechanism of formation. Our first experiments used monoclonal antibodies to keratins and an associated protein (vimentin), to look at the cells that compose a pterygium. These experiments demonstrated that a pterygium is the result of an abnormal limbal basal epithelial stem cell that moves onto Bowmans layer and brings about the dissolution of this layer. More importantly, these data showed that the clear corneal epithelial cells in front of the pterygium also contained these abnormal limbal cells, which we named the pterygium cell. This demonstrated that when a pterygium is removed, a wide area of what appears to be normal epithelium must be removed to inhibit reoccurrence of the growth. Later experiments using expressed sequence tag analysis of an un-normalized unamplified complementary DNA library from surgically removed pterygia were compared with normal cornea and confirmed the role of the epithelial cells in this growth. The gene expression studies also showed that genes involved in cellular migration are stimulated, and this led to studies on polyamine analogs as inhibitors of pterygial migration. Immunohistochemical studies with antibodies to matrix metalloproteinases (MMPs) showed that it is the pterygium cell that produces the MMPs that dissolve Bowmans layer resulting in the growth stimulation of stromal fibroblasts. This led to experiments on the use of MMP inhibitors to inhibit the growth of pterygia.

MLH1 and MSH2 expression in pterygia.

Purpose: Pterygia have been reported to share some of the genetic defects seen in cancers, including microsatellite instability (MSI). We examined pterygia for the presence of proteins typically missing or defective in adenocarcinomas with MSI. We also performed microsatellite analysis on DNA from pterygia to test for instability in the size of the microsatellites, using markers conventionally used to characterize MSI in tumors (Bethesda convention markers). Methods: We examined 13 pterygia by immunohistochemistry for MLH1 and MSH2, 2 proteins involved in DNA mismatch repair. In addition, we amplified the pterygial DNA with primers specific for 5 Bethesda markers (BAT25, BAT26, D2S123, D5S346, and D17S250). Results: MLH1 staining was present at low levels in the basal cells of the cornea and migrating limbal cells of the pterygia. MSH2 staining was present in basal and in maturing epithelial cells of the cornea and migrating limbal cells of pterygia. We observed no reproducible examples of MSI or loss of heterozygosity (LOH). Conclusions: We were unable to confirm the presence of MSI and LOH by using the markers we examined. MSH2 staining appeared to be normal in pterygia. MLH1 staining was present but in reduced amounts compared with that seen in the conjunctiva.