The effects of in vivo B-cell depleting therapy on ex-vivo cytokine production

Abstract

In renal transplantation, IL-17 production by T-cells might be dependent on the presence of B-cells. Therefore, the effect of in vivo B-cell depletion on ex-vivo IL-17 production was investigated. Twenty patients undergoing living-donor renal transplantation were recruited from a larger cohort of patients participating in a randomized, double-blind trial. All patients were allocated to a single intra-operative dose of either placebo or rituximab (375 mg/m(2)) added to the standard immunosuppressive therapy. Blood was collected at baseline, at one day, and at one month after surgery. The healthy kidney donors also gave blood at baseline. Peripheral blood mononuclear cells were stimulated ex-vivo in different manners (heat killed Candida albicans yeast, heat killed Staphylococcus aureus, or αCD3αCD28 coated beads), to address the role of B-cells in ex-vivo cytokine responses. The concentration of monocyte- and T-cell-derived cytokines (IL-1β, IL-6, TNF-α, IFN-γ, IL-17 and IL-22) was measured in supernatants. Of the 20 recruited patients, 13 received treatment with rituximab and 7 received placebo. In all patients, IL-17 was produced by CD4-positive, γδTCR-negative cells. After stimulation, there was no difference between patients and healthy controls in ex-vivo production of IL-17 or other cytokines. In all patients there was a general decrease of monocyte- and T-cell-derived cytokines after transplantation, except for IL-17. There was no difference between patients who received rituximab and patients who received placebo. A single dose of rituximab treatment added to standard immunosuppressive therapy in renal transplant patients did not influence the production of IL-17 or other monocyte- or T-cell derived cytokines after ex-vivo stimulation.

Trial registration: ClinicalTrials.gov NCT00565331.

Keywords: B-cell; Cytokines; Interleukin 17; PBMC; RA; Renal transplantation; SD; peripheral blood mononuclear cell; rheumatoid arthritis; standard deviation.