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A. Corell, R. Reinoso, M. Cuevas, R. Quispaya, E. Castellanos, S. Villarón, R. Martín, M. Martino, I. Fernández, M. Calonge; Flow Cytometric Analysis of Conjunctival Cells in Dry Eye Patients and Healthy Donors. Invest. Ophthalmol. Vis. Sci. 2010;51(13):4864.
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© ARVO (1962-2015); The Authors (2016-present)
To characterize conjunctival epithelial cells and intraepithelial lymphocytes (IELs) obtained by brush cytology (BC) from patients with dry eye disease (DED) and healthy donors.
BC samples were obtained from the inferior fornix in 23 patients with evaporative DED (ev-DED) (2 visits) and from superior tarsal conjunctiva in 11 patients with aqueous-deficient DED (ad-DED). Also, BC samples were obtained from 20 healthy controls for each DED type or conjunctival zone,and from an aditional area: superior bulbar conjuctiva. Flow cytometric analysis (FC) was performed to characterize cell lineage (epithelial cells and IELs subtypes: total T cells, helper T cells, cytotoxic T cells, B cells, and NK cells), cell viability, and DNA content.
Lymphocytic infiltration (CD45+ leukocytes) was significantly increased in ad-DED. However, no significant differences in any lymphocyte subset were observed between DED patients and healthy individuals. In ev-DED, a switch between CD4 and CD8 IELs was found when comparing diagnosis and post-treatment visits. When different conjunctival zones were compared in normal subjects, a higher T/B ratio and a lower TH/TC ratio were found in superior tarsal compared to inferior conjunctiva. Regarding cell viability, two different cell populations were constantly observed: a more numerous population of larger, more complex, and less viable cells and a less frequent population of smaller, less complex, and more viable cells. There were significant differences in the percentage of viable, apoptotic, and dead cells between DED and controls and between the anatomical area on healthy donors. Cell cycle assay revealed a lower cell proliferative capacity (S+G2/M) in both groups of DED patients than in healthy subjects and higher mean proliferative index in superior tarsal than in inferior conjunctiva.
Immunophenotyping and viability of BC-recovered cells show variations depending upon the anatomical zone with specific gradients from superior to inferior conjunctiva. DED patients, mainly from the ad-DED type, showed increased IELs, more epithelial damage, and a decreased cell proliferative capacity than healthy individuals.
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