Production of interleukin-6 and granulocyte-macrophage colony-stimulating factor by murine iris and ciliary body explants.
OBJECTIVE
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.
Massachusetts General Hospital-Harvard Cutaneous Biology Research Center, Department of Dermatology, Boston.
T L
TL
Knisely
S
S
Grabbe
R
R
Nazareno
R D
RD
Granstein
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002924:Ciliary Body / Q000276:immunology*; D046508:Culture Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis*; D007375:Interleukin-1 / Q000096:biosynthesis; D015850:Interleukin-6 / Q000096:biosynthesis*; D007498:Iris / Q000276:immunology*; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL
D007375:Interleukin-1; D015850:Interleukin-6; D016178:Granulocyte-Macrophage Colony-Stimulating Factor