Expression of iNOS mRNA and Effect of NO on IL-3 Production in Dendritic Epidermal T Cell

Seo, Seong Jun; Ro, Byung Kwan; Shin, Bong Ju; Hong, Chang Kwun

Abstract

BACKGROUND: Stimulation of iNOS with subsequent release of NO may play a critical role in pathophysiology or host defense reactions. NO can be produced by several cell types that reside in the skin. Increasing data indicate that NO serves as an autocrine and paracrine mediators in diverse and complex cellular processes throughout the skin.
OBJECTIVE
The aim of the present study was to elucidate whether dendritic epidermal T cell can express iNOS and generate NO. In addition to that, this study was to evaluate whether NO may play a certain role in IL-3 production of activated dendritic epidermal T cell.
METHODS
iNOS mRNA was isolated by RT-PCR at 12, 24, 48, 72 hours after stimulation and NO was measured by the Griess method. IL-3 levels were assessed with a basic cell proliferation bioassay system using supernatants from T cells cultured in 96 well plates. Units of IL-3 in the culture supernatants were measured using the cytokine-dependent DA-1 cell line.
RESULTS
1. NO generation was increased by UVB 100mJ/cm2 irradiation, concanavalin-A and lipopolysaccharide stimulation in dendritic epidermal T cell. The amount of NO was highest in LPS. 2. The expression of iNOS was increased by UVB 100mJ/cm2 irradiation, concanavalin-A and lipopolysaccharide stimulation in dendritic epidermal T cell. Induction of iNOS revealed peak at 72 hours after concanavalin-A and lipopolysaccharide stimulation, but in UVB irradiation that of iNOS was not observed except at 48 hours. 3. NO may play a role in IL-3 production in activated dendritic epidermal T cell.
CONCLUSION
The results strongly suggest that the expression of iNOS and NO production in dendritic epidermal T cell are upregulated by UVB, concanavalin-A and lipopolysaccharide, and NO may play a role as mediator in IL-3 production of activated dendritic epidermal T Cell as autocrine manner.