Novel model of innate immunity in XXXX .

@entity1 @entity32 @entity9504 @entity267 @entity3335 @entity19 @entity281 @entity687 @entity597
The @entity267 functions as the major refractive interface for vision and protects the internal eye from insult. Current understanding of innate immune responses to @entity281 derives from a synthesis of in vitro and in vivo analyses. However, monolayer cell cultures and @entity19 models do not accurately duplicate all aspects of innate immunity in @entity1 @entity1 . Here, we describe a three-dimensional culture system that incorporates @entity1 cells and extracellular matrix to more completely simulate the @entity1 @entity267 for studies of @entity281 . @entity1 corneal stromal fibroblasts were mixed with type I collagen in 3- m pore size transwell inserts, and overlayed with Matrigel to simulate a @entity1 corneal stroma and epithelial basement membrane. These were then infected with a @entity267 -tropic adenovirus, and exposed on their inferior side to leukocytes derived from @entity1 peripheral blood. Subsequent analyses were performed with histology, confocal microscopy, ELISA, and fluorescence-activated cell sorting (FACS). @entity687 , a neutrophil chemokine shown previously as the first cytokine induced in @entity281 of @entity1 corneal cells, increased upon @entity9504 of facsimiles in a dose-responsive fashion. @entity3335 -positive cells infiltrated infected corneal facsimiles in a sub-Matrigel location, possibly due to @entity687 colocalization with heparan @entity597 , a Matrigel constituent. Cellular infiltration was significantly inhibited by treatment with chemical inhibitors of p38 MAPK and Src kinase, both constituents of a signaling cascade previously suggested to regulate @entity32 after @entity9504 . FACS analysis determined that both virus and corneal fibroblasts were necessary for the induction of leukocyte migration into the facsimiles. The corneal facsimile, literally a @entity267 in a test tube, permits mechanistic studies on @entity1 tissue in a highly tractable system.
