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Published on: February 21, 2018
Production of interleukin-6 and granulocyte-macrophage colony-stimulating factor by murine iris and ciliary body
T L Knisely1, S Grabbe, R Nazareno
1Massachusetts General Hospital-Harvard Cutaneous Biology Research Center, Department of Dermatology, Boston.
Insights
Murine iris-ciliary body explants produce proinflammatory cytokines, including interleukin-6 (IL-6) and granulocyte-macrophage colony-stimulating factor (GM-CSF). These cytokines may influence the ocular immune microenvironment.
Area of Science:
- Ophthalmology
- Immunology
- Cell Biology
Background:
- The iris-ciliary body (I-CB) is a key ocular tissue involved in immune regulation.
- Understanding cytokine production by I-CB is crucial for comprehending ocular immune responses.
Purpose of the Study:
- To investigate the capacity of murine I-CB explants to generate cytokines with proinflammatory properties.
- To identify specific cytokines produced by I-CB tissue in vitro.
Main Methods:
- Murine I-CB explants were cultured in vitro.
- Culture supernatants were analyzed for cytokine production using thymocyte costimulation assays and ELISA.
- Key cytokines assessed included IL-1, IL-2, IL-3, IL-6, TNF-alpha/beta, and GM-CSF.
Main Results:
- I-CB explants produced significant thymocyte costimulatory activity, attributed to IL-6 and IL-1.
- IL-6 was detected at 402.7 pg/ml, and TNF-alpha/beta at approximately 2.0 U/ml.
- Significant amounts of GM-CSF (15.8 U/ml) were detected, while IL-2 and IL-3 were not found.
Conclusions:
- Normal I-CB tissue harbors cells that produce IL-6, GM-CSF, and IL-1.
- The proinflammatory nature of these cytokines and their role in antigen-presenting cell function suggest their importance in maintaining ocular immune homeostasis.
Purpose:
To examine the ability of murine iris-ciliary body explants to produce cytokines with proinflammatory activities.
Methods:
Supernatants derived from murine iris-ciliary body (I-CB) tissue explants cultured (four per well in 1 ml medium) in the presence of indomethacin were analyzed for the production of IL-1, IL-2, IL-3, IL-6, tumor necrosis factor-alpha/beta (TNF alpha/beta) and granulocyte-macrophage colony-stimulating factor (GM-CSF). Analysis was performed by thymocyte costimulation, growth factor-dependent cell proliferation, TGF-beta-sensitive mink lung epithelial cell proliferation, and enzyme-linked immunosorbent assays (ELISA).
Results:
Supernatants from I-CB explants cultured in vitro for 24 hours contained significant thymocyte costimulatory activity. This activity was fully neutralized by a combination of antisera to IL-1 and IL-6, and ELISA analysis confirmed that IL-6 was a significant component of the supernatant (402.7 pg/ml). TNF alpha/beta were also found in low concentrations (approximately 2.0 U/ml) by ELISA analysis, whereas IL-2 and IL-4 were not detectable. Significant amounts of GM-CSF (15.8 U/ml), but no IL-3, were detected.
Conclusions:
These results demonstrate that normal I-CB tissue contains cells capable of producing IL-6, GM-CSF, and IL-1. Because of the proinflammatory nature of IL-6 and IL-1, and the ability of GM-CSF, IL-1, and IL-6 to enhance functional capabilities of antigen presenting cells, regulation of the production of these cytokines may contribute significantly to the maintenance of the immunologic status of this regional site.
