Eleven ryegrass-sensitive patients were challenged weekly for 4 weeks with incremental doses of ryegrass allergen applied topically to one eye; a buffer was applied to the other eye. A clinical examination and tear-fluid cytology were performed before challenge and at 20 minutes, 1 hour, and 6 hours after challenge. A significant clinical reaction and neutrophil accumulation in the tear film occurred at 20 minutes. At 1 hour, a clinical response and tear cytologic reaction were present only at higher antigen concentrations. Six hours after antigen challenge, only the highest allergen concentration (320,000 BU/ml) produced a clinical late-phase reaction (LPR) (p less than 0.01) and tear cytologic change (presence of eosinophils and lymphocytes). Five nonryegrass-sensitive control subjects were unresponsive to a similar challenge. These results indicate that a conjunctival response to allergen challenge is dose dependent, that is, the higher the dose, the more likely an LPR will occur and that an LPR correlates with significant numbers of inflammatory cells in the tear film.