The Investigation Of Cytotoxic Effect Induced By Peripheral Blood Mononuclear Cells In The Patients With Psoriasis Vulgaris

Objective: Tumour necrosis factor (TNF) as a cytokine with many kinds of biological activities in vivo, has functions of killing tumour cells or inhibiting their growth, promoting inflammation and adjusting immunity. The biological activities of TNF are mainly showed by tumour necrosis factor-alpha (TNFα) which is classic TNF. According to the difference of its existence forms, TNFαhas been distinguished with secreted TNFα(sTNFα) and mebrane associated TNFα(MA-TNFα). Psoriasis (Ps) is a kind of common sickness with chronic and inflam characteristics. Since the Goerdt's finding of TNFαinducing endothelial cells activation in Ps vulgaris patients, more and more researches manifest the associativities between TNFαand psoriasis vulgaris. Our experiment is to investigate cytotoxic effect in L929 (mice fibroblasts) target cells by two forms of TNFαfrom peripheral blood monouclear cells (PBMC) in Ps vulgaris patients, and further illuminate the pathogenesy of sTNFαand MA-TNFαin Ps.Methods: We obtain monocytes from peripheral blood applying the methods of Ficoll density gradient centrifugation and adherency to 96-well plates, then prepare two type of TNFα――sTNFαand MA-TNFαusing different treatment with lipopolysaccharide (LPS) and acid buffer (AB). After culturing L929 with sTNFαor MA-TNFαfor 48 hours, we detect 570nm optical density (OD) to analyze the cytotoxic effect mediated by these two forms of TNFαin Ps patients and normals.