Purchase this article with an account.
P. Hamard, C. Blondin, C. Debbasch, L. Bensoussan, C. Baudouin, F. Brignole; In vitro Effects of Prostaglandin Analogs on Inflammatory Markers Expression by Human Conjunctival Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):4413.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: Conjunctival hyperemia has been reported in glaucoma patients treated with prostaglandin analogs and prostamides. We investigated whether these drugs would trigger in vitro expression of adhesion molecules or pro-inflammatory chemokines by human conjunctival cells. Methods: Conjunctival cells (5 x 104/ml, WKD cell line) were cultured in 6-well plates for 48 hr. Human recombinant TNFa (10 ng/ml) or IFNg (300 IU/ml) were added to the cells for the last 24 hr of culture. During the very last 4 hr, 0.03 % bimatoprost, 0.005% latanoprost, 0.004% travoprost, all diluted 1/100, or 10-5 M PGF2a were added to the cells previously stimulated or not with TNFa or IFNg. Non-treated cells were used as controls. Culture supernatants were harvested and assayed for IL-8 and MCP-1 by enzyme-linked immunosorbent assays. Adherent cells were trypsinized, stained by fluorescent antibodies and analyzed by flow cytometry for membrane expression of HLA-DR, ICAM-1 and PECAM-1 and for intracellular expression of the pro-inflammatory chemokines IL-8 and MCP-1. Results: Under basal conditions, conjunctival cells did not express HLA-DR, expressed low levels of the adhesion molecules ICAM-1 and PECAM-1, and did not secrete IL-8 and MCP-1. In response to TNFa stimulation, cells overexpressed both ICAM-1 (p<0.009) and PECAM-1 (p<0.03) and secreted IL-8 and MCP-1 (p<0.05). Similarly, expression of HLA-DR became detectable upon stimulation with IFNg (p<0.001). Upon stimulation with prostaglandin analogs, prostamides or PGF2a, the pattern of inflammatory marker expression remained unchanged. The response of cells treated simultaneously with a given drug and TNFa was similar to that of cells treated with TNFa alone. In the same way, stimulation of cells with IFNg alone or in the presence of various drugs resulted in a similar and significant increase in HLA-DR expression compared to stimulation with the drugs alone (p<0.001). Conclusions: These findings demonstrate that under our experimental conditions latanoprost, travoprost and bimatoprost did not display pro-inflammatory effects on resting human conjunctival cells, nor did they modify the response of these cells cultured in the presence of the pro-inflammatory cytokines TNFa or IFNg.
This PDF is available to Subscribers Only