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A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva
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A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva

Published on: July 5, 2017

Immune response in the conjunctival epithelium of patients with dry eye

Stefano Barabino1, Elisa Montaldo, Federico Solignani

  • 1Ocular Surface Research Center, Department of Neurosciences, Ophthalmology, and Genetics, University of Genoa, viale Benedetto XV 5, Genoa, Italy. barabino@schepens.harvard.edu

Experimental Eye Research
|July 27, 2010
PubMed

Insights

Dry eye patients show increased inflammation and immune cell changes in the conjunctiva. A new sample preservation method improves analysis of these cells, crucial for understanding dry eye disease.

Area of Science:

  • Ophthalmology
  • Immunology
  • Cell Biology

Background:

  • Dry eye disease is a common condition characterized by ocular surface inflammation.
  • The role of immune cell infiltration in the conjunctival epithelium of dry eye patients requires further elucidation.

Purpose of the Study:

  • To test the hypothesis that dry eye patients exhibit significant inflammation and lymphocyte infiltration in conjunctival epithelium.
  • To evaluate a novel method for preserving impression cytology samples for enhanced flow cytometry analysis.

Main Methods:

  • Impression cytology specimens were collected from 15 dry eye patients and 15 healthy controls.
  • Samples were preserved in either Phosphate Buffered Saline (PFA) or cell culture medium with foetal calf serum (FCS).
  • Flow cytometry was used to analyze cell phenotypes, including epithelial and immune cell markers (CK19, CD3, CD4, CD8, CD56, CD19, CD20, CD14, HLA-DR).

Main Results:

  • FCS-preserved samples yielded a statistically higher number of cells compared to PFA-preserved samples.
  • Dry eye patients showed a significant difference in the CD4/CD8 ratio and an increased number of CD14+ cells compared to controls.
  • Increased HLA-DR expression was observed specifically in CK19+ conjunctival epithelial cells of dry eye patients.

Conclusions:

  • Immune cells in the conjunctival superficial layer play a critical role in the pathogenesis of dry eye.
  • Preservation in FCS medium enhances the yield and analysis of conjunctival epithelial and immune cells via flow cytometry.
  • This method offers a promising approach for studying ocular surface inflammation in dry eye disease.

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