Abstract
Purpose: :
Neutrophils (PMN) are prominent infiltrating cells in acute herpetic keratitis as well as in chronic stromal keratitis. We evaluated PMN chemotaxis induced by supernatants obtained from corneal epithelial cells (HCE) and/or macrophages (THP-1 cells) treated with HSV components. We also studied induction of antiviral activities under these conditions.
Methods: :
HCE and THP-1 cells were infected with HSV-1, transfected with HSV DNA or treated with HSV-1 IgG complex(IC). After 8 to 24 hours incubation, supernatants were harvested and tested for their ability to chemoattract purified PMN (human CD16+cells) in a HTS 96 well plate (Corning NY) using an alamar blue bio-staining method. Supernatants of PMN treated for 24 hours with the HCE or THP-1 supernatants were assayed for IFN-, IFN-β, TNF- and antiviral activities.
Results: :
Chemotaxis occurred with the supernatants of HCE infected with HSV-1 or transfected with HSV DNA, but not with HSV-IC. Anti-IL-8 and anti-GRo- treatment inhibited chemotaxis. Anti-GM-CSF treatment also inhibited chemotaxis but to a lesser degree. Supernatants of macrophages treated with HSVDNA and HSV-IC also showed chemotactic activities and anti- IL-8, anti-GRo- and anti-GM-CSF acted similarly. PMN treated with HCE or macrophage supernatants have no IFN- or IFN-β. TNF- however, was found in the supernatants cultured with supernatants from HCE infected with HSV or transfected with HSVDNA and from macrophages transfected with HSV DNA or treated with HSV-IC. Weak antiviral activities were detected in some PMN supernatants as well.
Keywords: herpes simplex virus • keratitis • cytokines/chemokines