FRI0024 Multiplatform metabolomic study of cd4+ cells after methotrexate and infliximab treatment: focus on safety and tolerability profiles

Background Activated CD4+ cells comprise a large proportion of inflammatory cells involved in the development of rheumatoid arthritis (RA). Inflammatory cytokines, as tumor necrosis factor-a (TNFa), have also been implicated in the establishment and progression of joint destruction. Treatments for RA include synthetical and biological disease modifying anti-rheumatic drugs (DMARDs), as methotrexate (MTX) and TNFa-inhibitors, as infliximab. Cellular effects can be evaluated by metabolomics with measurement of ideally all endogenous metabolites. Objectives Evaluation in CD4+ cells, from a multiplatform metabolomic study, of MTX and infliximab treatment in order to investigate the metabolomic features. Methods Blood samples from 5 healthy controls were collected for metabolomic study. CD4+ cells were isolated by magnetic separation using CD4 MicroBeads from peripheral blood mononuclear cells, divided in 5 different groups and cultured for 24 hours: 1. CD4+ T cells as control, 2. CD4+ T cells treated with MTX at the dosage of 0.01 mM, 3. CD4+ T cells treated with MTX at the dosage of 0.1mM, 4. CD4+ T cells treated with infliximab at the dosage of 1 µg/ml, 5. CD4+ T cells treated with IgG1k at the dosage of 1µg/ml as control of Infliximab. A total of 25 cell samples were analysed using Metabolon’s standard extraction method. Results Ribulose was reduced with decrease in glycolysis and inhibition of phosphofructose kinase 1 (PFK1) by MTX (p