Tu1876 The Effects of Anti-TNF Agents on the Expansion of T Helper-Type 17 (TH17) Cells Driven by Lipopolysaccharide-Stimulated Monocytes

Patients and methods Monocytes and CD4+ cells were purified, by positive and negative selection, from peripheral blood mononuclear cells of healthy volunteers. CD4+CD45RO+ memory T cells were enriched from the CD4+ cell fraction by positive selection. A 1:1 ratio of monocytes and memory T cells was co-cultured for 7 days with CD3/CD28 Human T-Activator Dynabeads and 1 μg/mL LPS. Cells were cultured with and without 10 μg/mL anti-TNF agent. Subsequently, CD4+ cells were stained for intracellular Interferon g (INFg) and IL-17A, and analyzed by flow cytometry. IL-17A and IL-17F secretion was determined by ELISA.