Leukocyte Distribution in the Open Eye Tears of Normal and Dry Eye Subjects

Leukocytes accumulate in the eye with sleep, but little is known about the presence or absence of leukocytes in awake, open eye tears. This study sought to compare normal and dry eye subjects for daily variation in open eye leukocyte composition.

Ten normal subjects and nine dry eye subjects were enrolled. Subjects were trained for self-collection of tear samples using an ocular surface wash with 5 mL of phosphate buffered saline per eye. Subjects performed washes at awakening, between 8 and 9 am, between 11 am and 12 pm, and between 4 pm and 5 pm on four separate days. Leukocytes were isolated from the wash and were counted with a cell counter before staining with an anti-CD45 antibody and viability stain. Stained leukocytes were then analyzed via flow cytometry. Side scatter characteristics were used to distinguish granulocytes from lymphocytes. Results were interpreted both by time of wash as well as time from awakening.

At awakening, dry eye subjects had approximately twice as many recovered leukocytes and had a statistically significantly higher granulocyte-to-lymphocyte ratio as compared to normals. Leukocytes were rapidly cleared from the eye with a significant decrease in leukocyte counts at the 8 am time point as compared to awakening. Leukocyte counts across all open eye time points appeared to be consistent, with no differences between normal and dry eye subjects.

There is a low level, constitutively expressed population of leukocytes in the open eye tears of normal and dry eye subjects. Higher levels of granulocytes in dry eye disease subjects warrants further investigation into this population of cells, and their role in homeostasis and dysregulation.