Chemotactic activity of tears and bacteria isolated during adverse responses

Inflammatory processes are characterized by the dynamic influx of leukocytes. This leukocyte recruitment and activation is thought to be initiated by chemical signals including chemotactic factors. This study was designed to investigate the chemotactic activity in different tear types and bacteria isolated during adverse responses to contact lens wear. Chemotactic activity was determined by quantitating in vitro neutrophil migration using a microchemotaxis chamber. Results demonstrated that tears collected immediately after 8 hours sleep (P<0.001) and tears collected during adverse responses (P<0.001) showed significantly higher chemotactic activity compared to reflex tears. Specific neutralizing antibodies to IL-8, LTB4 and C5a were added to closed eye and adverse response tears. Pre-incubation of closed eye tears with antibodies to IL-8 showed a significant reduction in chemotactic activity (P<0.0001), whereas a significant reduction of PMN migration in adverse response tears was observed after pre-treatment with antibodies to LTB4 (P<0.0001). However no difference in chemotactic activity was observed after incubation with antibody to C5a or irrelevant antibody. Dot blots demonstrated that closed eye tears contained approximately 150 ng ml-1 IL-8 and adverse response tears contained 2 ng ml-1 IL-8. Most Gram negative bacteria isolated from contact lenses caused directed migration of PMNs. Addition of neutralizing antibody to LPS significantly abrogated the chemotactic activity of bacterial cells (P<0.001). Our findings provide evidence that IL-8 during eye closure, and bacterial chemotactic substances and LTB4 during contact lens induced adverse responses, are responsible for the recruitment of PMNs.