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Afsun Sahin, Didem Cosan, Cağri Oner, Hasan V. Gunes, Nilgun Yildirim, Hikmet Basmak; Regulation of pro-inflammatory cytokine secretion in human meibomian gland epithelial cells. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):3506.
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© ARVO (1962-2015); The Authors (2016-present)
Androgens are known to modulate the function of ocular surface and adnexal epithelial cells, suppress lipopolysaccharide(LPS)-induced proinflammatory responses in nonocular sites. However, there is limited data if androgens show this anti-inflammatory activity on ocular surface and adnexa. We hypothesize that human ocular adnexal epithelial cells have the ability to produce and release pro-inflammatory cytokine (IFN-gamma, TNF-alpha, IL-10, IL-1beta, IL-2, IL-4, IL-6, IL-8, VEGF-A) in response to lipopolysaccharide exposure. We also hypothesize the following: (1) Lipopolisaccharide binding protein (LBP) potentiates LPS-induced cytokine secretion by human meibomian gland epithelial cells; (2) dihydrotestosterone (DHT), a potent androgen, attenuates the immune effect of LPS. The objective of this study was to test our hypotheses.
Immortalized human meibomian gland epithelial cells (from David A.Sullivan, Schepens Eye Research Institute, Boston, MA) were cultured in keratinocyte serum-free medium until reaching 100% confluence. Cells were then exposed to DMEM/F12 medium with 10% BCS for 2 days, followed by an incubation in serum-free DMEM/F12 for 1 day. After this time period, cells were treated with vehicle or 15 ug/ml LPS (E. Coli, strain 0127:B8) for 6, 12 and 24 hours. Culture media were collected and analyzed for IFN-gamma, TNF-alpha, IL-10, IL-1beta, IL-2, IL-4, IL-6, IL-8, VEGF-A levels with Luminex technology.
Lipopolysaccharide stimulates time-dependent secretion of IL-1beta, IL-6, IL-8, and VEGF-A by human meibomian gland epithelial cells. This effect is potentiated by exposure to LPS-binding protein. This potentiation, in turn, is significantly reduced by cellular treatment with dihydrotestosterone.
Human meibomian gland epithelial cells have the ability to generate pro-inflammatory cytokine in response to LPS exposure. This proinflammatory process is modulated by LPS-binding protein and by dihydrotestosterone. When induced by appropriate stimuli, this cytokine production may have a role in the generation of inflammation in ocular surface disease.
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