In vivoeffects of cyclic administration of 15-deoxyspergualin on leucocyte function in patients with Wegener's granulomatosis

Abstract

Summary 15-Deoxyspergualin (DSG)is an alternative treatment modality for Wegener's granulomatosis (WG) patients refractory to conventional treatment. Nevertheless, it is unclear how DSG modulates disease activity in these patients. This study was conducted to investigate which parameters of adaptive and acquired immunity were influenced during two subsequent cycles of DSG treatment. Emphasis was put upon T cell and monocyte activation, neutrophil function and surface expression of proteinase-3 (PR-3). Anti-CD3/anti-CD28 and interleukin (IL)-15/IL-7-mediated T cell proliferation were assessed by fluorescence activated cell sorter (FACS) analysis using carboxyfluorescein succinimidyl ester (CSFE) labelling. Interferon (IFN)-g and IL-10 production were determined in the supernatants of these cultures by enzyme-linked immunosorbent assay. Monocyte activation was assessed in lipopolysaccharide (LPS)-stimulated whole blood, using tumour necrosis factor (TNF)-a as read-out. Neutrophil function was determined by measuring oxidative burst, chemotaxis and phagocytosis. T cell activation markers and PR3 expression were measured by FACS. All parameters were determined directly before and after each DSG cycle. Anti-CD3/anti-CD28-mediated T cell proliferation was reduced directly after DSG treatment. Directly before a subsequent cycle of DSG was started, T cell proliferation was increased. Similar findings were observed for IFN-g and IL-10 production by T cells. DSG did not influence IL-15/IL-7-mediated T cell proliferation. LPS-mediated TNF-a production was also impaired directly after DSG treatment. No influence on T cell activation markers, neutrophil function and surface PR-3 expression was observed in peripheral blood of these patients. Our data demonstrate that DSG influences T cell and monocyte activation in a reversible fashion. Although DSG causes neutropenia in these patients, it does not influence neutrophil function.

Deoxyspergualin in relapsing and refractory Wegener's granulomatosis

“…Dysregulated antigen presentation appears to be important in Wegener’s granulomatosis, and deoxyspergualin hinders the ability of monocytes and macrophages to act as antigen-presenting cells through inhibition of the production of oxygen free radicals and downregulation of MHC class II expression 31 32. Although treatment with deoxyspergualin leads to a marked reduction in neutrophil numbers, neutrophil function is not affected 33. The induction of neutropaenia may, however, be important as neutrophil depletion has been shown to be protective against the development of glomerulonephritis in a mouse model of ANCA-associated vasculitis 34.…”

Pharmacological Targeting of the Hsp70 Chaperone

“…Although DSG was originally identified as a potential anti-bacterial agent, it also has anti-tumor activity and it has found clinical use in organ transplantation and autoimmune diseases [177–179]. DSG prolongs survival time after allogenic graft and minimizes acute rejection.…”

Monitoring cell proliferation in vitro with different cellular fluorescent dyes

“…Some T cells after activation induce physiological apoptotic process [7]. Although commonly used in diagnoses, the cell tracer CSFE [8][9][10][11] emits fluorescence in the range frequently used for antibody labelling; therefore, it may be inconvenient in some studies. In those cases CellVue Claret [5] or CellTrace Violet labelling may be preferred.…”