|Title||COMPOSITION OF INFLAMMATORY CELLS SHED FROM THE OCULAR SURFACE|
|Author, Co-Author||Jie Zhou, Carolyn Begley, Graeme Wilson|
|Abstract|| PURPOSE. Overnight eye closure induces an accumulation of inflammatory cells, mostly PMNs. Inflammatory cells can be collected throughout the day using the method of contact lens cytology (CLC). The purpose of this study was to determine the immune activation of these inflammatory cells, and the number of T lymphocytes found in CLC collection from normal subjects.
METHODS. Ocular surface cells were collected using the method of CLC from 5 subjects upon eye opening in the morning and at 1:00p.m. the same day. Samples were pooled from both eyes of each subject, incubated with 0.1% BSA in PBS buffer, divided into 4 equal portions for analysis, and incubated with one of the following: FITC-conjugated monoclonal anti-human HLA-DR (labels immune-activated and Langerhans cells), FITC-conjugated monoclonal anti-human CD3 (labels T lymphocytes), monoclonal antibody to human common leukocyte antigen T200 followed by incubation with FITC-conjugated goat anti-mouse IgG (labels all inflammatory cells), or FITC-conjugated, isotype-matched non-specific mouse antibody (negative control). All samples were then fixed in 2% paraformaldehyde, counterstained with ethidium homodimer, and viewed by epifluorescent microscopy.
RESULTS. In samples collected upon eye opening, an average of 7.6% of the inflammatory cells were HLA-DR and 6.0% were CD3 positive, out of approximately 250 inflammatory cells (T200-positive). In eye opening samples, many cells were trapped in mucous threads, so the percent of stained cells may be an underestimate. Afternoon samples contained an average of 17.1% HLA-DR positive and 13.0% CD3 positive cells. Cell sizes ranged from 6-22 micron (HLA-DR positive) and 6-13 micron (CD3 positive). Both mononuclear and multinuclear cells were HLA-DR positive.
CONCLUSIONS. CLC collections include T lymphocytes and activated inflammatory cells, including PMNs. Larger (15-20 micron) HLA-DR positive cells in CLC collections may be Langerhans cells (cons
|Affiliation of Co-Authors||School of Optometry|