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Characterization of a Novel Human Organotypic Retinal Culture Technique
Published on: June 9, 2021
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Human retinal endothelial cells express functional interleukin-6 receptor.
Lisia Barros Ferreira1, Liam M Ashander1, Binoy Appukuttan1
1Flinders University College of Medicine and Public Health, Flinders Medical Centre, Rm 4E-431, Bedford Park, Adelaide, SA, 5042, Australia.
Journal of Ophthalmic Inflammation and Infection
|April 25, 2023
Summary
Human retinal endothelial cells express Interleukin-6 receptor (IL-6R), enabling classic IL-6 signaling. This finding challenges previous assumptions and has implications for treating non-infectious uveitis.
Area of Science:
- Ophthalmology
- Immunology
- Cell Biology
Background:
- Interleukin-6 (IL-6) drives inflammation in non-infectious uveitis.
- IL-6 signals via classic (IL-6 receptor dependent) and trans-signaling pathways.
- Vascular endothelial cells were thought to lack IL-6 receptor (IL-6R) expression.
Purpose of the Study:
- To investigate IL-6R expression in human retinal endothelial cells (HRECs).
- To determine if HRECs can utilize classic IL-6 signaling.
- To assess the functional impact of IL-6 on HRECs.
Main Methods:
- Reverse transcription-polymerase chain reaction (RT-PCR) to detect IL-6R transcripts.
- Flow cytometry to identify IL-6R protein expression.
- Real-time transcellular electrical resistance measurements to assess HREC monolayer integrity upon IL-6 stimulation.
Main Results:
- IL-6 receptor (IL-6R), membrane-bound IL-6R (mIL-6R), and soluble IL-6R (sIL-6R) transcripts were detected in primary HRECs.
- Flow cytometry confirmed intracellular IL-6R and surface mIL-6R expression in HRECs.
- IL-6 treatment significantly reduced HREC monolayer electrical resistance, indicating functional IL-6R activity.
Conclusions:
- Human retinal endothelial cells express functional IL-6R.
- Classic IL-6 signaling is possible in HRECs.
- These findings impact therapeutic strategies for IL-6-driven uveitis.
