Cytolytic effect of macrophages and polymorphonuc lear leukocytes treated with acidic fraction of bakers' yeast mannan against MM46 tumor cells in vitro.

Biological & Pharmaceutical Bulletin

Abe

Watanabe

et al. 2002

Self Cite

Kupffer cells (KC), 1) resident macrophages of the liver, comprise a large and functionally important part of the mononuclear phagocyte system. Interleukin 1 (IL-1) is a monokine with a broad spectrum of biological activities, and its effects on a variety of cell types have been established. There are several reports that suggest that IL-1 could serve as a therapeutic tool for patients and mice with bacterial and fungal infections. [2][3][4][5][6] There is also accumulating evidence that lipopolysaccharide (LPS)-stimulated KC produce IL-1 or IL-1-like factors.7-12) However, the role of KC, which could be an important source of these mediators, has not been completely evaluated to date.We have reported that an acidic mannan fraction (WAM025) of bakers' yeast confers significant protection against the ascites forms of Ehrlich, sarcoma 180, and Meth A tumors, 13) and Candida albicans infections 14) in mice. Until now, there have been few reports of the production of IL-1 from phagocytes by mannan, except for our previous report that polymorphonuclear leukocytes (PMN) in peritoneal exudate cells, but not macrophages, of mice administered WAM025 produced larger amounts of IL-1. 15) In this study, we describe the production of IL-1 activity by KC treated with WAM025 in vivo and in vitro to clarify further the antitumor and infection-protective mechanisms. MATERIALS AND METHODSMice Specific pathogen-free (SPF) male BALB/c mice, 5 to 6 weeks old, were obtained from the Shizuoka Laboratory Animal Center, Hamamatsu, Japan. Preparation of Mannan FractionsThe mannan subfractions were prepared as previously reported.16) Briefly, the bulk mannan fraction extracted from the parent bakers' yeast, a wild-type strain of the Saccharomyces cerevisiae cells, was fractionated by DEAE-Sephadex A-50 column chromatography, and the neutral and the strongly acidic fractions, which were eluted with water and 0.25 M NaCl (designated as WNM and WAM025, respectively), were used in this study.Treatment of Mice The mice were intraperitoneally (i.p.) administered the mannan fraction (WNM or WAM025) at a dose of 150 mg/kg/d for 5 d.Preparation of KC The KC were prepared by a method based on that of Hashimoto et al.17) Briefly, the liver was perfused with 0.2% Pronase E (Kaken Pharmaceutical), then the small fragments were digested for 20 min in the same solution at 37°C. After the incubation, 0.5 mg of deoxyribonuclease (DNase) type I (Merck) was added to digest the cellular debris. After 2 repeated 20 min-DNase treatments, the KC were then separated by Percoll (Pharmacia) gradient centrifugation (density 1.080). The KC were washed three times with Hanks balanced salt solution and suspended in 10% FBS-RPMI-1640 medium. The KC were estimated to be 98% viable on the basis of trypan blue dye exclusion. The KC preparations were judged to be 90 to 95% pure after peroxidase staining. Thymocyte Proliferation Assay IL-1 activity in culture supernatants was assayed by the enhancement of the thymocyte proliferation activity as described by Vacheron et al. ...

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Production of Interleukin-1 Activity of Kupffer Cells from Mice Treated with the Acidic Mannan Fraction of Bakers' Yeast.

Biological & Pharmaceutical Bulletin

Abe

Watanabe

et al. 2002

Self Cite

“…14) Preparation of the peripheral blood phagocytes (monocytes and PMNs) was conducted in accordance with the previous description.…”

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confidence: 99%

Role of Polymorphonuclear Leukocytes in the Resistance of Tumor-Bearing Mice against Candida albicans Infection

Biological & Pharmaceutical Bulletin

Kobayashi

Sakai

et al. 2004

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Opportunistic fungal infection occurring frequently in a compromised host is one of the most important problems in modern medicine, because of the difficulty of treatment and the frequency with which such infections progress to a lifethreatening condition. There are many papers reports on the increased susceptibility of tumor-bearing hosts to systemic fungal infections. [2][3][4][5][6] The protective activity of allogeneic sarcoma 180 tumor-bearing mice against Candida albicans and Aspergillus fumigatus infections has been reported in our previous papers. [7][8][9][10][11] It has also been reported that there is a significant delay in the lethal response to C. albicans in mice bearing Lewis lung carcinoma. 12) However, the mechanisms of such an effect have yet to be fully elucidated. We reported that the number of peripheral blood polymorphonuclear leukocytes (PMNs) dramatically increased in sarcoma 180 tumor-bearing mice. [7][8][9] We also investigated the producibility of the granulocyte-macrophage colony-stimulating factor from the tumor cells. 9,10) We thought that this phenomenon should be investigated not only in allogeneic tumor-bearing mice but also in syngeneic tumor-bearing mice. In the present study, to demonstrate further the importance of PMNs in the resistance of tumor-bearing mice against C. albicans, we conducted a protection assay against the infection using allogeneic (sarcoma 180) and syngeneic (Meth A fibrosarcoma [Meth A] and MM 46 mammary carcinoma [MM46]) solid tumor-bearing mice over the course of 3 weeks after tumor cell transplantation. Furthermore, we investigated in detail the involvement of PMNs from tumor-bearing mice as the likely mechanism of such protection. MATERIALS AND METHODS Mice and TumorsMale 6-to 8-week-old ddY, BALB/c, and C3H/He mice were obtained from the Shizuoka Laboratory Animal Center, Hamamatsu, Japan. Sarcoma 180 (1ϫ10 6 ), 3-methylcholanthrene-induced Meth A fibrosarcoma (Meth A, 1ϫ10 6 ), and MM 46 mammary carcinoma (MM46, 3ϫ10 6 ) were transplanted subcutaneously in the ddY, BALB/c, and C3H/He mice, respectively. Cyclophosphamide (CY)-treated mice were prepared as follows: 16 d after tumor transplantation, the mice were injected intraperitoneally with a single dose of 15, 50, or 200 mg/kg of CY (Cytoxan, Shionogi Co., Ltd., Osaka, Japan).Fungus The C. albicans NIH A-207 strain was kindly donated by Dr. T. Shinoda, Department of Microbiology, Meiji College of Pharmacy, Tokyo, Japan, and was maintained on Sabouraud agar slants. For this study, the fungus was cultured in a Sabouraud liquid medium which was shaken at 27°C for 48 h.Infection-Protection Assay One, 2, and 3 weeks after tumor cell transplantation, the mice were inoculated intravenously with 2ϫ10 6 viable C. albicans cells. The mice bearing only the solid tumor began dying by week 4; therefore reliable data could not be obtained beyond 3 weeks. In the CY experiment, 4 d after CY administration, the mice were infected intravenously with the fungal cells. The mortality rate of the mice was recorded for...

“…[1][2][3][4] In order to identify the competent cells in acidic mannan (WAM025)-treated peritoneal exudate cells (PEC), we carried out an in vitro cytolysis assay by macrophages and polymorphonuclear leucocytes (PMN) separated from the PEC by the Percoll gradient centrifugation technique. 5) The results indicated that WAM025-treated macrophages were able to display a marked cytolytic effect, while PMN from the same source did not show the effect. Because the chemical entity involved in the cytolysis of tumor cells by WAM025-treated macrophages remained to be identified, we attempted to characterize the cytolytic factor elicited in mouse macrophages by the administration of the yeast mannans.…”

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confidence: 93%

Correlation between Tumor Cell Cytotoxicity and Serine Protease Activity of Peritoneal Macrophages from Mice Treated with Bakers' Yeast Mannans

CHEMICAL & PHARMACEUTICAL BULLETIN

Osuga

Suzuki

et al. 1987

Self Cite

Peritoneal exudate cells (PEC) from C3H/He mice given bakers' yeast mannans, WNM and WAM025, showed marked cytolytic activity against MM46 tumor cells in vitro. Because the cytotoxicity of the PEC was strongly inhibited by actinomycin D, it was assumed that the cytolytic factor was a proteinous material(s). Of the macrophages and polymorphonuclear leucocytes (PMN ) separated from the PEC, only the former cells were found to display cytolytic activity. Quinacrine, a phospholipase A2, inhibitor, and sodium azide, a myeloperoxidase inhibitor, did not reduce the cytotoxicity of macrophages from mice treated with acidic mannan (WAM025). In contrast, diisopropylfluorophosphate, a serine protease inhibitor, inhibited the cytolysis of the MM46 target cells by WAM025-treated macrophages. Chymostatin, a chymotrypsin inhibitor, and elastatinal, an elastase inhibitor, also inhibited the cytolytic effect. It is therefore concluded that serine proteases secreted into the medium from the macrophages activated by mannans participated in the cytolytic destruction of neoplasmic cells.Macrophage activation for nonspecific tumoricidal activity against target tumor cells can be induced by various immunopotentiators. In. our previous reports, it was found that mannan of bakers' yeast was able to enhance the immunological response of mice to transplanted tumor cells and to lethal infection with pathogenic microbes.1-4) In order to identify the competent cells in acidic mannan (WAM025)-treated peritoneal exudate cells (PEC), we carried out an in vitro cytolysis assay by macrophages and polymorphonuclear leucocytes (PMN) separated from the PEC by the Percoll gradient centrifugation technique.5) The results indicated that WAM025-treated macrophages were able to display a marked cytolytic effect, while PMN from the same source did not show the effect. Because the chemical entity involved in the cytolysis of tumor cells by WAM025-treated macrophages remained to be identified, we attempted to characterize the cytolytic factor elicited in mouse macrophages by the administration of the yeast mannans.The relevancy of serine protease release to tumoricidal activity of macrophages has been reported in mice treated with BCG6,7) and Nocardia rubra cell-wall skeleton.8) However, the real cytolytic factors of antitumor polysaccharide-treated macrophages have not been elucidated to date, although many reports have been published on identifying various lytic factors produced by cytolytically activated macrophages such as oxygen intermediates,9,10) lysosomal enzymes,11,12) proteases,6,7) complement component C3,13) tumor necrotic factor,14) prostaglandins, 15) and myeloperoxidase.16) Therefore, the present study was designed to investigate the possible role of serine proteases in the cytolysis of target tumor cells by antitumor mannan-induced macrophages.