ifn-γ sirna effectively knocked down ifn-γ gene expression and reduced cytokine secretion in peripheral blood mononuclear cells of patients with autoimmune hepatitis

Background: autoimmune hepatitis (aih) is an inflammatory liver disorder that commonly affects women. the t cells and one of their major products, ifn-γ, are critically involved in the pathogenesis of aih. therefore, targeting of ifn-γ can probably be thera- peutically useful while avoiding the long-term side effects of conventional immunosuppressive therapy. rna interference, provides an ideal way to achieve this purpose. thus, the aim of this study was to investigate the effect of ifn-γ-sirna on ifn-γ expression in human peripheral blood mononuclear cells of patients with aih. methods: in order to evaluate the anti-cytokine therapy with ifn-γ-sirna, the pbmcs of aih patients were cultured and transfected with ifn-γ-sirna. after validation of transfection efficiency, the effects of gene silencing were tested with quantitative real-time polymerase chain reaction and intracellular flow cytometry. results: the efficiently transfected cells, with the targeted ifn-γ-sirna, without affecting cell viability showed a strong gene knock- down. the ifn-γ gene expression was significantly decreased in transfected cells (p < 0.05). moreover, flow cytometric analysis confirmed the decrease of the intracellular ifn-γ protein level after sirna transfection. conclusions: collectively, the results of the present study suggested that modifying the cytokine profile without inducing apopto- sis using sirna-based technology could be a promising tool for therapeutic intervention in t cells-dependent inflammatory diseases like aih.