Production of Interleukin‐10 by Combining a Granulocyte and Monocyte Adsorption Carrier With Ulinastatin

Abstract

Interleukin (IL)-10 is an anti-inflammatory cytokine mainly produced by monocytes and is essential for the induction of anti-inflammatory intestinal macrophages with macrophage colony-stimulating factor (M-CSF). Thus, IL-10- and M-CSF-rich conditions in colonic tissues seem to contribute to the improvement of pathological conditions in patients with inflammatory bowel diseases (IBD). We have already reported that ulinastatin, a serine protease inhibitor, increases M-CSF production during granulocyte/monocyte (GM) adsorption to cellulose acetate (CA) beads (carriers for Adacolumn therapy). However, the effects of ulinastatin on IL-10 production have not been clarified. The aim of the present study was to clarify the effects of ulinastatin on IL-10 production during GM adsorption by in vitro experiments. Peripheral blood was divided into four groups: (Control) no ulinastatin added, no contact with CA beads; (1) no ulinastatin added, contact with CA beads; (2) ulinastatin added, no contact with CA beads; and (3) ulinastatin added, contact with CA beads. After incubation, IL-10 in the plasma was measured. Compared with the level in the Control group, plasma IL-10 was significantly higher only in group 3, in which ulinastatin was added in the presence of CA beads, but did not increase in the absence of CA beads. These results suggest that ulinastatin synergistically increases IL-10 production with monocyte adsorption stimuli. By increasing not only M-CSF but also IL-10, a combination of ulinastatin and Adacolumn therapy may improve clinical efficacy for the treatment of IBD in terms of the induction of anti-inflammatory intestinal macrophages.

Effect of Cellulose Acetate Beads on the Release of Transforming Growth Factor‐β

“…Therefore, the reduction of TGF‐β release via CA beads might be due to this reduction of Th17‐cell differentiation, which may be one of the mechanisms of clinical efficacy of GMA in patients with IBD. Moreover, we also revealed that ulinastatin not only had the ability to increase the release of anti‐inflammatory cytokines, such as IL‐1ra and IL‐10 , but that it could also decrease TGF‐β release (Fig. ) in combination with GM and platelet adsorption to CA beads.…”

“…We have already reported that cytokine release (e.g. IL‐1ra, HGF, IL‐10, and TNF‐α) is augmented by GM adsorption to CA beads . Thus, we expected that TGF‐β release would be augmented by incubation with CA beads because platelets, which produce TGF‐β, are also adsorbed (Fig.…”

“…After incubation, plasma was separated by centrifugation to measure TGF ‐β and IL ‐6. Data for the IL ‐1 receptor antagonist ( IL ‐1ra) and IL ‐10 were obtained from previous reports . The decrease ratio (black circle) of TGF ‐β and the increase ratios (white circle) of IL ‐1ra, IL ‐10, and IL ‐6 were calculated as described in Materials and Methods.…”

“…These devices comprise a column filled with 2‐mm cellulose acetate (CA) beads that act as carriers for adsorptive leukocyte apheresis. We previously reported that contact between peripheral blood and CA beads induces the release of not only anti‐inflammatory cytokines, including IL‐1 receptor antagonist (IL‐1ra), hepatocyte growth factor (HGF), and IL‐10 , but also pro‐inflammatory cytokines, such as tumor necrosis factor‐α (TNF‐α) . These cytokines are released via the stimulation of GM adsorption to CA beads.…”

Involvement of Extracellular Signal‐Regulated Kinase in Leukocyte Adsorption‐Induced Production of Hepatocyte Growth Factor

“…We previously reported that contact between peripheral blood and cellulose acetate (CA) beads used as carriers for granulocyte and monocyte (GM) adsorptive apheresis (GMA) increases plasma HGF concentration (3), through unknown signaling pathways that are involved in HGF production in GM adsorption to CA beads. On the other hand, we also reported that ulinastatin, a serine protease inhibitor, increased the production of anti-inflammatory cytokines such as interleukin (IL)-1 receptor antagonist and IL-10 in combination with GM adsorption to CA beads (4). In this study, we investigated the effect of ulinastatin on HGF production in GM adsorption and elucidate the signaling pathway involved.…”