Limitations of an ocular surface inflammatory biomarker in impression cytology specimens
| Autor: | Annette John-Baptiste, Rolla Yafawi, Frederick Sace, Mira Ko |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Adult
Pathology medicine.medical_specialty genetic structures Context (language use) Biology Toxicology medicine.disease_cause Flow cytometry medicine HLA-DR Humans Inflammation Reproducibility medicine.diagnostic_test Epithelial Cells Impression cytology HLA-DR Antigens General Medicine Assay sensitivity Flow Cytometry eye diseases Biomarker (medicine) Dry Eye Syndromes sense organs Irritation Conjunctiva Biomarkers |
| Zdroj: | Cutaneous and Ocular Toxicology. 32:46-53 |
| ISSN: | 1556-9535 1556-9527 |
| Popis: | A number of ocular conditions, such as dry eye, are associated with inflammation on the surface of the eye leading to irritation and ocular pain. Many drugs such as chemotherapeutics, beta blockers, angiotensin-converting enzymes and so forth also cause dry eye but currently there are no validated ocular surface biomarkers available.We evaluated sample stability, assay sensitivity, reproducibility and overall performance of impression cytology (IC) utilizing the cellular surface biomarker human leukocyte antigen DR-1 (HLA-DR) as an ocular surface inflammatory biomarker by flow cytometry in a fit-for-purpose validation study. Additionally, subjects classified as normal or having various degrees of dry eye were evaluated to determine if HLA-DR could demonstrate a clear separation between normal and dry eye samples.The assay demonstrated high dynamic range detecting a broad range of fluorescent intensities in healthy donors. Additionally, inter, intra and stability assay results demonstrated strong concordance and low variability. Overall CV% for both assays were less than 25% for all measured parameters. However, high variability was observed for donor samples assayed beyond day 10 post IC sample collection (4.2-110.8 CV%).HLA-DR expression demonstrated a progressive increase in patients with mild to severe levels of dry eye disease providing sufficient evidence it is sensitive enough to monitor inflammatory effects of dry eye when coupled with additional biomarkers and/or methodologies such as cytokine analysis or ICAM-1. This biomarker can be used to monitor ocular surface disorders in patients and to evaluate potential treatment options during drug development. Although our results demonstrate this methodology is reproducible for routine evaluation, limitations around sample integrity exist.The ocular cell surface inflammatory biomarker, HLA-DR coupled with impression cytology is a simple non-invasive robust, specific and reproducible assay that can be utilized to measure inflammatory infiltrates on the surface of the eye in IC samples less than 10-days old. |
| Databáze: | OpenAIRE |
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