Suzuki, Yasuo; Yamada, Akihiro; Aoki, Hiroshi; Osamura, Aisaku; Nakamura, Kentaro; Yoshimatsu, Yasushi; Hosoe, Nobuo; Takada, Nobuo; Tsuda, Yukio; Shirai, Koji (2008). W1254 The Decreased TLR2 Expression and Elevated IL-8 Production On Peripheral Leukocytes in Patients with Active Ulcerative Colitis: the Modulation of the Leukocyte Sensitivity to Pgn By Granulocyte and Monocyte Apheresis. Gastroenterology, 134(4), A-665–. doi:10.1016/S0016-5085(08)63105-4 

BACKGROUND & AIM: Mucosal lesions of active ulcerative colitis (UC) are characterized by acute inflammatory cell (polymorphonuclear granulocyte, PMN) infiltrate and ulceration. Granulocyte and monocyte apheresis (GMA) using a column device, Adacolumn, selectively adsorbs and removes leukocytes from the peripheral blood, and has been reported to be effective in patients with active UC. The aim of this study was to investigate the expression of toll like receptor 2 (TLR2) on PMNs and monocytes in patients with active UC as compared to the level in healthy subjects and see the impact of GMA on TLR2. METHODS: Fortyeight patients with active UC, Clinical Activity Index (CAI, Lichtiger index) ≥5, were included in this study. Patients received several GMA sessions (maximum 10 sessions), and CAI ≤ 3 was considered remission. Blood samples were obtained from patients at each GMA session and also from healthy subjects (n=14), after obtaining informed consent. The expression of TLR2 was investigated by flowcytometry. In several samples, IL-8 production by cultured blood upon stimulation with peptidoglycan (PGN, a ligand for TLR2) was measured (n=21). RESULTS: An average of CAI at the entry was 11.0 (n=48), and remission rate by GMA was 56.3%(27/48). At the baseline, the expression of TLR2 on PMN of patients with active UC was significantly less than the level in healthy subjects (p<0.001). Further, the improvement in CAI was accompanied by a trend towards normalization of TLR2 on PMN (up-regulation, p<0.05) together with a significant fall (P<0.01) of plasma IL-8 level. However, as for the patients with still active symptoms (CAI>5) in course of GMA therapy, the decreased TLR2 on PMN and the elevated release of IL-8 from PGN-stimulated leukocytes were kept on. By exposing blood to the column carriers In Vitro, TLR2 expression on the cell surface of PMN was decreased, but increased within the cellular of PMN. Similarly, in the blood on column outflow of GMA, the TLR2 expression on PMN and IL-8 production by PGN-stimulation were significantly decreased (TLR2:P<0.001, IL-8:P<0.001), indicating that leukocyte sensitivity to PGN is weakened by GMA column. CONCLUSION: The expression of TLR2 on PMN in patients with active UC is significantly decreased compared with remission status or healthy subjects. In connection with the decreased TLR2 expression, the IL-8 production by PGN-stimulated leukocytes is elevated in patients with active UC. These changes seem to reflect PMN activation based on UC disease activity. GMA or exposure of blood to GMA carriers is associated with down-modulation of the TLR2 expression on PMN and also leukocyte sensitivity to PGN

https://www.gastrojournal.org/article/S0016-5085(08)63105-4/pdf

Grit Waitz ¹, Sebastian Petermann, Stefan Liebe, Joerg Emmrich, Wolfgang Ramlow

Dig Dis Sci. 2008 Sep;53(9):2507-15. doi: 10.1007/s10620-007-0168-8. Epub 2008 Feb 6.

The influence of the granulocyte/monocyte apheresis (GMCAP) on cell populations participating in mechanisms of tolerance, e.g. dendritic cells (DCs), is still not very clear. In a first step, we aimed to investigate changes in the DC population of patients suffering from ulcerative colitis (UC) (n = 13) compared to healthy subjects (n = 9). In a second step, we studied the changes in peripheral DCs in a small group of patients with active UC before and after Adacolumn apheresis (n = 7). For this purpose, plasmacytoid and myeloid DCs and their maturation markers CD40, CD80, and CD86 were measured using four-color flow cytometry in the peripheral blood. After apheresis, and in acute flare-ups, we identified a significantly lower number of lymphocytes, plasmacytoid, and myeloid DCs. In conclusion, the additional removal of peripheral DCs by GMCAP, which otherwise would contribute to the inflammatory process in the gut, may lead to a higher tolerogeneic status towards luminal antigens.

https://pubmed.ncbi.nlm.nih.gov/18253828/

https://link.springer.com/article/10.1007%2Fs10620-007-0168-8

Ritsuko Yanaru-Fujisawa, Takayuki Matsumoto, Shotaro Nakamura, Shuji Kochi, Mitsuo Iida, Futoshi Kohda, Minako Hirahashi, Takashi Yao, Ryuichi Case Reports Inflamm Bowel Dis. 2005 Aug;11(8):780 DOI: 10.1097/01.mib.0000172558.39767.b7

Our case and the case of Kanekura et al⁸ suggest that circulating leukocytes may play an important role in the pathogenesis of PG and that GCAP in combination with corticosteroids may be a promising strategy for intractable PG. Furthermore, as has been the case for active UC, GCAP may be a choice for severe pouchitis. An accumulation of data with respect to the effect of GCAP on pouchitis seems to be warranted.

https://pubmed.ncbi.nlm.nih.gov/16043996/

https://academic.oup.com/ibdjournal/article/11/8/780/4685895

Yukihiko Kawasaki ¹, Junzo Suzuki, Shigeo Suzuki, Hitoshi Suzuki

J Pediatr Gastroenterol Nutr. 2004 Oct;39(4):422-5. doi: 10.1097/00005176-200410000-00021.

Generally, UC is associated with intervals of acute exacerbation and the administration of corticosteroids is effective in bringing about a clinical remission (¹). Corticosteroids are not always effective even in doses over 1 mg/kg/day. In addition, the long-term use of corticosteroids often causes serious side effects such as growth retardation, glaucoma, hormonal disturbance, peptic ulcer, liver dysfunction and psychologic problems. Alternative treatment for active UC may be necessary to avoid the clinical problems associated with corticosteroid therapy. In recent years, leukocytapheresis (LCAP) and granulocytapheresis (GCAP) using a leukocyte removal filter has been found effective in some cases of adults with inflammatory bowel disease (^2,3). However, there have been few reports concerning the efficacy of LCAP and GCAP for UC in childhood (⁴).We report two children with severe steroid-dependent UC in whom LCAP with leukocyte removal filter was used in treatment. LCAP therapy was safe and effective in two children with refractory UC and allowed discontinuation of corticosteroid therapy with an improvement in quality of life. Prospective studies of this therapy will be needed to clarify the role of LCAP in treatment of childhood UC.

https://pubmed.ncbi.nlm.nih.gov/15448435/

https://journals.lww.com/jpgn/Fulltext/2004/10000/Leukocytapheresis_with_Leukocyte_Removal_Filter.21.aspx

Katsuya Hiraishi ¹, Yuji Takeda, Noriyuki Shiobara, Hiromu Shibusawa, Fumie Jimma, Nobuhito Kashiwagi, Abby R Saniabadi, Masakazu Adachi, Ther Apher Dial. 2003 Jun;7(3):334-40.

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 degrees 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 FcgammaR 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.

https://pubmed.ncbi.nlm.nih.gov/12924609/

Abby R Saniabadi ¹, Hiroyuki Hanai, Ken Takeuchi, Kazuo Umemura, Mitsuyoshi Nakashima, Taro Adachi, Chikako Shima, Ingvar Bjarnason, Robert Lofberg, Ther Apher Dial. 2003 Feb;7(1):48-59.

Apheresis has been recognized both economically and therapeutically as a novel approach for the treatment of inflammatory diseases, and certain others, which respond poorly to drug therapy. This report is about Adacolumn, an adsorptive carrier based granulocyte and monocyte apheresis device with a volume of 335 mL, filled with about 220 g of cellulose acetate beads of 2 mm diameter as the column adsorptive carriers. Pre- and post-column leukocyte counts have shown that the carriers adsorb about 65% of granulocytes, 55% of monocytes and 2% of lymphocytes from the blood in the column. Additionally, after apheresis, there is a marked decrease in inflammatory cytokines (TNF-alpha, IL-1beta, IL-6 and IL-8) produced by blood leukocytes, together with down-modulation of L-selectin and the chemokine receptor CXCR3. Adacolumn has been used to treat patients with rheumatoid arthritis, ulcerative colitis and HIV infection. Typical apheresis sessions have been 4-10, at a frequency of one or two sessions per week. Treatment of patients with Adacolumn has been associated with very promising efficacy and safety data. Accordingly, in Japan, Adacolumn has been approved by the Ministry of Health for the treatment of ulcerative colitia. Furthermore, Adacolumn met the required quality and safety standards for medical devices and received an EC certification (CE-mark) from TUV in 1999. However, although Adacolumn carriers are very efficient in depleting excess and activated granulocytes and monocytes/macrophages, the clinical efficacy associated with Adacolumn apheresis cannot be fully explained on the basis of reducing granulocytes and monocytes per se. Hence, a long lasting effect on inflammatory cytokine generation, chemokine activities or immunomodulation is likely, but the precise mechanisms involved are not fully understood yet.

https://pubmed.ncbi.nlm.nih.gov/12921115/

Priscilla Biswas Barbara Mantelli Alberto Beretta Clinical Immunology 109 (2003) 355–358 DOI: 10.1016/j.clim.2003.07.001 

Therapeutic purging of myeloid cells (monocytes and granulocytes) (MYP) has been proposed as a treatment of severe inflammatoryconditions like ulcerative colitis and rheumatoid arthritis. Although direct purging of inflammatory cells contributes to its efficacy, the precise mechanism of action is still unclear. We have tested MYP in a pilot study on 12 patients with chronic HIV infection, of whom 6 underwent MYP. Three/6 MYP patients and none of the controls displayed a strong and long-lasting decrease of cells expressing CXCR3,a major chemokine receptor responsible for trafficking of inflammatory cells. In these three patients, the number of circulating CD4 T cells increased during treatment. The data provide a rational for the use of MYP as a therapeutic tool acting via the modulation of immune cell trafficking

https://europepmc.org/article/med/14697751

Nobuhito Kashiwagi ¹, Minoru Nakano, Abby R Saniabadi, Masakazu Adachi, Toshikazu Yoshikawa

Inflammation 2002 Aug;26(4):199-205. doi: 10.1023/a:1016523914161.

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-alpha 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-alpha 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.

https://pubmed.ncbi.nlm.nih.gov/12184634/

Nobuhito Kashiwagi ¹, Kazuhito Sugimura, Hirobumi Koiwai, Hironori Yamamoto, Toshikazu Yoshikawa, Abby R Saniabadi, Masakazu Adachi, Takashi Shimoyama

Dig Dis Sci. 2002 Jun;47(6):1334-41. doi: 10.1023/a:1015330816364.

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-alpha, interleukin-1beta, interleukin-6, and interleukin-8) production by leukocytes, neutrophil chemotaxis, down-regulation of leukocyte adhesion molecule (L-selectin) and neutrophil adhesion to interleukin-1beta-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.

https://pubmed.ncbi.nlm.nih.gov/12064810/