Abstract
Purpose :
The primary objective of the study is to compare the number of transcripts of inflammatory mediators on the ocular surface of healthy individuals with no ocular symptomatology to the number of transcripts in patients who have been diagnosed with the ocular manifestation of GvHD and have not received therapy for ocular GvHD to determine whether biomarker testing might be used to detect early onset of the disease.
Methods :
Eleven GvHD patients and six age and sex-related controls were evaluated. OSDI questionnaire and clinical tests (corneal and conjunctival staining, and Schirmer test) were performed. Conjunctival cells were collected using an EYEPRIM™ device 5 minutes after instilling a topical ophthalmic anesthetic. The device was placed on the bulbar conjunctiva near the limbus and held for 5 seconds. After collecting the cells on the membrane, the membrane was placed in a tube containing storage medium and stored at 4°C. The RNA was isolated; cDNA synthesis was performed and samples were analyzed using real-time polymerase chain reaction (PCR) assay. The number of transcripts of human leukocyte antigen-DR (HLA-DRA), interferon-gamma (IFN-γ), and interleukin-6 (IL-6) genes were measured. The student t-test for independent samples was used to compare means between groups.
Results :
HLA-DRA, IFN-γ and IL-6 transcripts levels were increased in GvHD patients compared to controls (1830.03 vs 189.44 (p=0.04), 64.46 vs 32.93 (p=0.004) and 43.52 vs 3.23 (p=0.03), respectively). There was a positive correlation between corneal fluorescence and lissamine green staining (R=0.79 and p=0.006).
Conclusions :
Analysis of ocular surface for markers of inflammation such as, HLA-DRA, IFN-γ and IL-6 can be used diagnostic tool to detect early onset of the disease as well as preventative test post BMT, for which there is no standard diagnostic test. Further studies are needed to use the level of mediators as diagnostic markers of the disease.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.