Masakazu Adachi

Immunomodulatory Effects of Granulocyte and Monocyte Adsorption Apheresis as a Treatment for Patients with Ulcerative Colitis

Our aim was to understand the mechanism of immunological changes associated with the use of an adsorptive-type extracorporeal device (Adacolumn) that has been developed for selective adsorption of granulocytes and monocytes/macrophages from peripheral blood of patients with active ulcerative colitis. The column is filled with carriers (G-1 beads) that have a diameter of 2 mm and are made of cellulose diacetate. In peripheral blood treated with the G-1 beads or peripheral blood from patients with active ulcerative colitis following granulocyte and monocyte adsorption apheresis, a significant suppression of proinflammatory cytokines (tissue necrosis factor-α, interleukin-1β, interleukin-6, and interleukin-8) production by leukocytes, neutrophil chemotaxis, down-regulation of leukocyte adhesion molecule (L-selectin) and neutrophil adhesion to interleukin-1β-activated endothelial cells were observed. Furthermore, after granulocyte adsorption therapy, the number of CD10-negative premature granulocytes increased, indicating increased turnover of these cells in the circulation. Our observations suggest that selective granulocyte and monocyte adsorption is associated with modified peripheral blood leukocyte function favorable to patients with ulcerative colitis and possibly other autoimmune disorders which reflect leukocyte hyperactivity.

Studies on the mechanisms of leukocyte adhesion to cellulose acetate beads: an in vitro model to assess the efficacy of cellulose acetate carrier-based granulocyte and monocyte adsorptive apheresis.

Abstract: Granulocyte and monocyte adsorptive apheresis (GMA) using a column filled with cellulose acetate (CA) beads (carriers) has been associated with a significant clinical efficacy in patients with rheumatoid arthritis and ulcerative colitis. To obtain further understanding on the mechanisms of disease modification by cellulose acetate‐carrier‐based GMA, in the present study, we investigated the mechanisms of granulocyte and monocyte adhesion to CA beads following exposure of human peripheral blood to the carriers at 37°C for up to 60 min under controlled conditions. Cellulose acetate beads selectively adsorbed granulocytes, monocytes, CD19+ (B cells) and CD56+ (NK cells) lymphocyte subpopulations. The granulocyte and monocyte adsorption was inhibited by heat‐inactivated plasma and EDTA, indicating that the adsorption was plasma protein (immunoglobulin, complement) and calcium dependent. Accordingly, granulocyte and monocyte adsorption was markedly enhanced by coating the carriers with IgG. Similarly, C3b was adsorbed onto the CA beads as a marker of complement activation. The results indicated that IgG and active complement fragments mediated leukocyte adhesion to CA beads via the FcγR and/or leukocyte complement receptor like CR3. Additionally, CA beads induced loss of expression of TNF receptors on CD16+ granulocytes and CD14+ monocytes, but not on CD3+ lymphocytes. In conclusion, CA beads might be an appropriate biomaterial for inducing extracorporeal immunomodulation as a treatment for auto‐immune diseases which are associated with pathological leukocyte activity.

Anti-inflammatory effect of granulocyte and monocyte adsorption apheresis in a rabbit model of immune arthritis.

In active rheumatoid arthritis, large numbers of granulocytes and macrophages are found in the inflamed joints. These leucocytes can promote inflammation and tissue injury by releasing inflammatory cytokines, proteinases and oxygen derivatives. To see if granulocyte and monocyte (GM) depletion produces anti-inflammatory effect, GM adsorption apheresis was performed in rabbits with immune arthritis by using a column (Adacolumn) filled with cellulose diacetate beads (G-1 beads) as adsorptive carriers which selectively adsorb CD11b positive GMs. Injection of ovalbumin into the knee joints of ovalbumin-sensitized rabbits caused a marked increase in peripheral blood leucocytes, joint swelling, increased granulocyte adhesion to G-1 beads and elevated TNF-α production by peripheral blood mononuclear cells (PBMC). When rabbits received a 60 min adsorption apheresis, there was suppression of CD11b positive leucocyte infiltration into the joint and reduced joint swelling (P < 0.01) compared with controls. Additionally, there was a significant (p < 0.01) suppression of TNF-α production by PBMC in the post column blood. These results suggest that GM depletion may serve as a non-pharmacological strategy to modify inflammatory disorders.

A novel method for determination of α1,6fucosyltransferase activity using a reducing oligosaccharide from egg yolk as a specific acceptor

A new method for determination of α1,6fucosyltransferase activity has been described. Recently, the disialyl-biantennary undecasaccharide was prepared in high yield from egg yolk [(1996), Carbohydr Lett 2: 137–42]. By treatment of this oligosaccharide with neuraminidase and β-galactosidase, we readily obtained an asialo-agalacto-biantennary heptasaccharide (GlcNAcβ 1,2Manα1,6[GlcNAcβ1,2Manα1,3]Manβ1,4GlcNAcβ1,4GlcNAc). Using this asialo-agalacto-oligosaccharide as an acceptor, fucosyltransferases from human plasma and extracts of various human hepatoma cell lines were assayed in the presence of GDP-[3H]fucose. The reaction mixture was applied to a column of GlcNAc-binding, Psathyrella velutina lectin coupled gel. All the fucosylated acceptor were bound to the column which was eluted with 50 mM GlcNAc. Structural analyses revealed that only the innermost GlcNAc residue of the acceptor was fucosylated through an α1,6-linkage, and the oligosaccharide prepared could be used as a specific acceptor for α1,6fucosyltransferase. The present method was used to screen plasma α1,6fucosyltransferase in several patient groups, and significantly elevated activities were found in samples from patients with liver diseases, including chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma.

MK615, A Compound Extract from the Japanese Apricot "Prunus mume" Inhibits In vitro Cell Growth and Interleukin-8 Expression in Non-small Cell Lung Cancer Cells

The Japanese apricot “Prunus mume,” which is also known as the Ume fruit in Japan, is a centuries-old traditional Japanese medicine, and it is a commonly consumed food. MK615, a compound extract from Ume fruits, has been shown to have anti-tumor and anti-inflammatory effects. In this study, we assessed the effects of MK615 on the in vitro growth of nine non-small cell lung cancer (NSCLC) cell lines and the HBEC4 immortalized bronchial epithelial cell line. While MK615 inhibited the in vitro cell growth of the majority of the NSCLC cell lines, the growthinhibitory effects varied among the cell lines, and some cell lines exhibited MK615 resistance. In the H1299 and H157 NSCLC cell lines that are highly sensitive to MK615, the induction of autophagy was observed after MK615 treatment. In addition, cell-cycle analysis showed that MK615 increased the proportion of cells in the G0-G1 phase in H1299 and H157 cells. In H1792 cells that overexpress IL-8, MK615 down-regulated IL-8 expression at the mRNA and protein levels in a dose-dependent manner. These results suggest that MK615 has multiple anti-tumor activities including the inhibition of cell proliferation, autophagy induction, G0-G1 cell cycle arrest and the downregulation of IL-8 expression in NSCLC cells.