Tumor-specific cytocidal and immunopotentiating effects of relatively low molecular weight products derived from the basidiomycete, Agaricus blazei Murill.

Fujimiya Y; Suzuki Y; Katakura R; Ebina T

Division of Immunology, Miyagi Cancer Center Research Institute, Natori, Jp

ABSTRACT

 Currently, some natural herbal extracts are believed to have a marked tumoricidal effect and low toxicity for normal tissues. We investigated the effect of relatively low molecular weight products extracted from the basidiomycete, Agaricus blazei Murill, on MethA tumor cell growth with the aim of producing synthetic derivatives based on these products. Inoculation of the low molecule fraction (LM) into the primary tumor of a two-tumor model resulted in the marked inhibition of the tumor, not only in the right flank, but also in the non-injected left flank. Chromatographic purification and physicochemical characterization showed the main tumoricidal activity to be located in a low molecule fraction-3 (LM-3), containing alpha-1,4-glucan-beta-1,6- glucan complex with an average molecular weight of 20 kDa. A11 LM fractions and crude ATF showed in vitro selective cytotoxicity for MethA tumor cells, having no effect on normal cells. Serum levels of immunosuppressive acidic protein (IAP) in mice receiving LM fractions, particularly LM-3, significantly increased, indicating the possible activation of granulocytes. We speculate that the inhibition of the distant tumor might be due to the increased migration of granulocytes, enhanced by the effect of extract injections at the primary tumor site.

Anticancer Res,  1999 Jan-Feb;19(1A):113-8 - PMID: 10226531 UI: 99243118

Cancer Immunol Immunother; 46(3):147-59 1998

Selective Tumoricidal Effect of Soluble Proteoglucan Extracted from the Basidiomycete, Agaricus blazei Murill, Mediated Via Natural Killer Cell Activation and Apoptosis

Fujimiya Y, Suzuki Y, Oshiman K, Kobori H, Moriguchi K, Nakashima H, Matumoto Y

Division of Immunology, Miyagi Cancer Center Research Institute, Natori, Japan.

ABSTRACT

We have isolated a novel type of natural tumoricidal product from the basidiomycete strain, Agaricus blazei Murill. Using the double-grafted tumor system in Balb/c mice, treatment of the primary tumor with an acid-treated fraction (ATF) obtained from the fruit bodies resulted in infiltration of te distant tumor by natural killer (NK) cells with marked tumoricidal activity. As shown by electrophoresis and DNA fragmentation assay, the ATF also directly inhibited tumor cell growth in vitro by inducing apoptotic processing; this apoptotic effect was also demonstrated by increased expression of the Apo2.7 antigen on the mitochondrial membranes of tumor cells, as shown by flow-cytometric analysis. The ATF had no effect on normal mouse splenic or interleukin-2-treated splenic mononuclear cells, indicating that it is selectively cytotoxic for the tumor cells. Cell-cycle analysis demonstrated that ATF induced the loss of S phase in MethA tumor cells, but did not affect normal splenic mononuclear cells, which were mainly in the G0G1 phase. Various chromatofocussing purification steps and NMR analysis showed the tumoricidal activity to be chiefly present in fractions containing 1-->4)-alpha-D-glucan and (1-->6)-beta-D-glucan, present in a ratio of approximately 1:2 in the ATF (molecular mass 170 kDa), while the final purified fraction, HM3-G (molecular mass 380 kDa), with the highest tumoricidal activity, consisted of more than 90% glucose, the main component being (1-->4)-alpha-D-glucan with (1-->6)-beta branching, in the ratio of approximately 4:1.

Cancer Immunol Immunother; 46(3):147-59 1998

Antitumor Effects of a New Polysaccharide-protein Complex (ATOM) Prepared from Agaricus blazei (Iwade strain 101) "Himematsutake" and Its Mechanisms in Tumor-bearing Mice.

Ito H, Shimura K, Itoh H, Kawade M

Department of Pharmacology, Mie University School of Medicine, Japan.

ABSTRACT

The antitumor activity of the i.p. or p.o. administration of polysaccharide-protein complex, ATOM (antitumor organic substance Mie) prepared from cultured mycelia of Agaricus blazei (Iwade strain 101) "Himematsutake" examined against four kinds of established mouse tumors. ATOM was highly effective at the doses of 10 and 20 mg/kg/day x 10 on subcutaneously implanted Sarcoma 180 in mice, and was also active against Ehrlich ascites carcinoma, Shionogi carcinoma 42 and Meth A fibrosarcoma at doses of 50 and 100 mg/kg/day x 10. ATOM has no direct cytotoxic action on tumor cells in vitro. Thus the tumor growth-inhibitory effect of ATOM is apparently due to immunological host-mediated mechanisms. The number of peritoneal macrophages, the phagocytosis of polystyrene latex beads and the proportion of the third component of complement (C3)-positive fluorescent cells were increased in the mice treated with ATOM. These results suggest that the macrophage activation and alterations of the C3 are necessary for the induction of an antitumor effect of ATOM.

Anticancer Research; 17(1A):277-84 1997

Inhibitory action of a (1 fwdarw 6)-beta-D-Glucan-protein complex (FIII-2-b) isolated from Agaricus blazei Murill ("Himematsutake") on Meth A fibrosarcoma-bearing mice and its antitumor mechanism.

Itoh-H; Ito-H; Amano-H; Noda-H

Lab. Marine Biochemistry, Faculty Bioresources, Mie Univ., Tsu, Mie 514, Japan

ABSTRACT

The effects of F III-2-b (Agaricus blazei Murill polysaccharide) with or without 5-fluorouracil (5-FU) on immune responses were investigated in Meth A tumor-bearing and normal mice. The i.p. administration of F III-2-b (10 mg/kg/day times 30) moderately inhibited the growth of Meth A tumor cells implanted s.c. in mice. Development of implanted tumors was strongly inhibited by the combination of FIII-2b and 5-FU. The picryl chloride-induced delayed type hypersensitivity (PC-DTH) response in mice was depressed after the implantation of tumor and treatment with 5-FU. FIII-2-b restored the suppression of PC-DTH by 5-FU, but did not increase the PC-DTH of normal mice. FIII-2-b not only enhanced the degree of spleen cell-mediated sheep red blood cells (SRBC) hemolysis (quantitative hemolysis of SRBC), the indexes of the spleen and thymus, and the number of spleen cells but also restored the suppressive effect of 5-FU. In the group receiving F III-2-b, the percentages of splenic Thy1.2-, L3T4- and asialo GM1-positive cells were significantly increased as compared with the tumor-bearing mice treated with saline. Furthermore, the L3T4+/Lyt2+ ratio showed a tendency to increase, and the Lyt2+/Thy1.2+ ratio was markedly decreased. These results suggest that the antitumor effect of F III-2-b may be correlated with the changing pattern of the Thy1.2-, L3T4- and asialo GM1-positive cells.

Biological Abstracts Vol. 99, Iss. 3, Ref. 36601.

Japanese Journal of Pharmacology 66(2): 265-271 - 1994

Antimutagenic and bactericidal substances in the fruit body of a Basidiomycete Agaricus blazei

Osaki-Y; Kato-T; Yamamoto-K; Okubo-J; Miyazaki-T

Tokyo Coll. Pharmacy, 1432-1 Horinouchi, Hachioji, Tokyo 192-03, JAP

ABSTRACT

The fruit body of a Basidiomycete Agaricus blazei, Jun-17 (Himematsutake) was extracted with hexane and chloroform-methanol (2:1, v/v), and the antimutagenic effect of the extracts was examined using an Ames/Salmonella/microsome assay. Both extracts of Agaricus inhibited the mutagenicity of benzo(a)pyrene(B(a)P). The hexane extract was purified by silica gel column chromatography and high performance liquid chromatography (HPLC), and linoleic acid was isolated as a main substance having antimutagenic activity. Fr. IIa, IIb, IIc and IIb, which reduced the number of His+ revertant colonies induced by B(a)P, were separated from the chloroform-methanol extract by silica gel column chromatography and HPLC. An antimutagenic substance in Fr. IIa was linoleic acid. From Fr. IIb, a bactericidal, not antimutagenic, substance was isolated and identified as 13-hydroxy cis-9, trans-11-octadecadienoic acid (13ZE-LOH). Antimutagenic substances in Fr. IIc and IId were not purified. The possible source and mechanism of formation of 13ZE-LOH are discussed.

Biological Abstracts Vol. 98, Iss. 7, Ref. 93597

Yakugaku Zasshi 114(5): 342-350 - 1994

Antitumor activity and some properties of water-soluble polysaccharides from "Himematsutake", the fruiting body of Agaricus blazei Murill.

MIZUNO-T; HAGIWARA-T; NAKAMURA-T; ITO-H; SHIMURA-K; SUMIYA-T; ASAKURA-A

Faculty Agriculture, Shizuoka Univ., Ohya, Shizuoka 422, Jpn 

ABSTRACT

Polysaccharides extracted from Himematsutake, the fruiting body of Agaricus blazei Murill with hot water were fractionated and purified by ethanol percipitation, ion-exchange chromatography, gel-filtration, affinity chromatography, etc. A total of 17 polysaccharide samples thus obtained were given an antitumor activity test (Sarcoma 180/mice i.p. p.o. method) and traces of their activities through the fractionation and purification processes were found. FI-0-a-beta, FA-1-a-alpha, FA-1-a-beta, and FA-2-b-beta, were obtained as water soluble polysaccharides fractions having great antitumor activities. Analyses of physico-chemical properties and IR- and NMR-spectra of these active fractions showed that their main components were: FI-0-a-beta, (1 fwdarw 6)-; (1 fwdarw 3)-beta-D-glucan; FA-1-a-alpha, acidic (1 fwdarw 6)-; (1 fwdarw 4)-alpha-D-glucan; FA-1-a-beta, acidic (1 fwdarw 6)-; (1 fwdarw 3)-alpha-D-glucan; and FA-2-b-beta, acidic RNA-protein complex.

Japan Journal of Pharmacology; 66(2):265-71 1994

AGRICULTURAL AND BIOLOGICAL CHEMISTRY 54(11): 2897-2906 - 1990

Antitumor activity and some properties of water-insoluble hetero-glycans from "Himematsutake", the fruiting body of Agaricus blazei Murill.

MIZUNO-T; INAGAKI-R; KANAO-T; HAGIWARA-T; NAKAMURA-T; ITO-H; SHIMURA-K; SUMIYA-T; ASAKURA-A

Faculty Agriculture, Shizuoka Univ., Ohya, Shizuoka 422, Jpn 

AGRICULTURAL AND BIOLOGICAL CHEMISTRY 54(11): 2897-2906 - 1990

ABSTRACT

After extraction of a hot-water-soluble polysaccharide (FI) from the fruiting bodies of Himematsutake (Agaricus blazei Murill), water-insoluble polysaccharides were obtained by successive extraction with 1% ammonium oxalate solution (FII), 5% sodium hydroxide solution (FIII and FIV), 20% sodium hydroxide solution (FV), and 5% lithium chloride-dimethylacetamide solution (FVI) in that order. These water-insoluble fractions were further fractioned by ethanol precipitation, gel-filtration, etc. Polysaccharides, polysaccharide-protein complexes, and chitin substances thus obtained were assayed for their antitumor activities using the Sarcoma 180/mice i.p., p.o. method. The heteroglycan-protein complexes, FII-a, -b and -c, obtained from FII had weak antitomor activities. A remarkable antitumor activity was found in a glycoprotein, FIII-2-b, fractionated and purified from FIII. The polysaccharide portion of this polysaccharide-protein complex (polysaccharide, 50.2% and protein, 43.3% each on a weight basis) consisted of (1 fwdarw 6)-7b-D-glucan, and its protein portion was rich in Asx, Glx, Ala, Leu, and Pro. A high antitumor activity was found in a xyloglucan-protein complex, FIV-2-b, fractionated and purified from FIV. Antitumor activity was found also in a glucosylan, FV-2-a, obtained from FV. No significant antitumor activity was found in a chitin substance, FVI.

Biological Abstracts Vol. 91, Iss. 7, Ref. 75458.

Antitumor activity and some properties of water-soluble polysaccharides from "Himematsutake", the fruiting body of Agaricus blazei Murill.

MIZUNO-T; HAGIWARA-T; NAKAMURA-T; ITO-H; SHIMURA-K; SUMIYA-T; ASAKURA-A

Faculty Agriculture, Shizuoka Univ., Ohya, Shizuoka 422, Jpn 

ABSTRACT

Polysaccharides extracted from Himematsutake, the fruiting body of Agaricus blazei Murill with hot water were fractionated and purified by ethanol percipitation, ion-exchange chromatography, gel-filtration, affinity chromatography, etc. A total of 17 polysaccharide samples thus obtained were given an antitumor activity test (Sarcoma 180/mice i.p. p.o. method) and traces of their activities through the fractionation and purification processes were found. FI-0-a-beta, FA-1-a-alpha, FA-1-a-beta, and FA-2-b-beta, were obtained as water soluble polysaccharides fractions having great antitumor activities. Analyses of physico-chemical properties and IR- and NMR-spectra of these active fractions showed that their main components were: FI-0-a-beta, (1 fwdarw 6)-; (1 fwdarw 3)-beta-D-glucan; FA-1-a-alpha, acidic (1 fwdarw 6)-; (1 fwdarw 4)-alpha-D-glucan; FA-1-a-beta, acidic (1 fwdarw 6)-; (1 fwdarw 3)-alpha-D-glucan; and FA-2-b-beta, acidic RNA-protein complex.

Biological Abstracts Vol. 91, Iss. 7, Ref. 75457.

AGRICULTURAL AND BIOLOGICAL CHEMISTRY 54(11): 2897-2906 - 1990

Fractionation and antitumor activity of the water-in-soluble residue of Agaricus blazei fruiting bodies.

KAWAGISHI-H; INAGAKI-R; KANAO-T; MIZUNO-T; SHIMURA-K; ITO-H; HAGIWARA-T

Dep. Agric. Chem., Fac. Agric., Shizuoka Univ., 836 Ohya, Shizuoka 422, Jpn 

ABSTRACT

Some polysaccharide-containing materials were successively extracted from the fruiting bodies of Agaricus blazei with aqueous ammonium oxalate and sodium hydroxide, fractionated, and assayed for antitumor activity. From chemical analyses and n.m.r. data, it was concluded that the most active fraction, FIII-2-b, was comprised of protein and a (1 fwdarw 6)-beta-D-glucan.

Biological Abstracts Vol. 88, Iss. 2, Ref. 17651

CARBOHYDRATE RESEARCH 186(2): 267-274 - 1989.

Formolysis of a Potent Antitumor (1-6)-beta-D-Glucan-Protein Complex from Agaricus blazei Fruiting Bodies and Antitumor Activity of the Resulting Products.

Kawagishi H, Kanao T, Inagaki R, Mizuno T, Shimura K, Ito H, Hagiwara T, Nakamura T

Dept. Applied Biol. Chem, Fac. Agric., Shizuoku Univ., 836 Ohya, Shizuoku 422, Jp

ABSTRACT

A potent antitumor (1--6)-beta-D-glucan-protein complex, isolated from the water-insoluble residue of Agaricus blazei fruiting bodies, was formulysed in order to separate the complex into its polysaccharide and protein components and assay the antitumor activity of each component. A pure glucan obtained did not exibit strong activity. The results suggest that the protein component is necessary for the potent activity of the complex.

Agricultural and Biological Chemistry; 54(11):2889-2896 1990

Screening of Host-Mediated Antitumor Polysaccharides by Crossed Immunoelectrophoresis Using Fresh Human Serum

Shimura K, Ito H, Hibasami H

Institute of Laboratory Animals, Mie University School of Medicine, Tsu, Japan.

ABSTRACT

On crossed immunoelectrophoresis, human serum C3 (the third component of complement) converted by antitumor polysaccharides (ATSO [antitumor polysaccharide oral], AB-P [Agaricaus blazei polysaccharide], GU-P [Grifora umbellata polysaccharide], PS-K [polysaccharide Kureha] and zymosan) moved faster than native C3, appearing as the 3rd peak. The ratio of height of the 3rd peak to the alpha 2-macroglobulin (alpha 2-M) peak was linearly proportional to the dose of ATSO. At the dose of 500 micrograms/ml antitumor polysaccharides, the ratios were higher than 0.76, and the ratios for the serum treated with polysaccharide of no antitumor activity (dextran and gum arabic) were less than about 0.52. This ratio readily determined in vivo can be used as a measure for the antitumor activity of polysaccharides.

Japan Journal of Pharmacology; 33(2):403-8 1983

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Books about mushrrons & medicine

Chang, S.-t., J. Buswell & S-w. Chiu (eds). 1993. Mushroom Biology and Mushroom Products. Proceedings of the First international Congress. Hong Kong: The Chinese University Press. 370 pg.

Hobbs, C., 1995. Medicinal Mushrooms: an Exploration of Tradition, Healing and Culture. Botanica Press. 252 pg.

Jones, K., 1995. Shiitake: The Healing Mushroom. Healing Arts Press.

Jones, K., 1997. Cordyceps: Tonic Food of Ancient China. Sylvan Press.

Lee, W. & H & J.A. Friedrich, 1997. Medicinal Benefits of Mushrooms. Keats Publishing.

Royse, D.J. (ed). 1996. Mushroom Biology and Mushroom Products. Proceedings of the Second International Congress. PennState Univ Press. 581 pg.

Stamets, P. 1993. Growing Gourmet and Medicinal Mushrooms by Paul Stamets. Cultivation techniques and growth requirements for 25 types of mushrooms.

Wagner, H. and Proksch, A., 1985. Immunostimulatory Drugs of Fungi and Higher Plants, Ecorpomic and Medical Plant Research, Academic Press, New York.

 Willard, T., 1990. Reishi Mushroom: Herb of Spiritual Potency and Medical Wonder. Sylvan Press. 167 pg.

Publications about mushroons & health

Beinfield, H., 1997. Medicinal mushrooms: help yourself to a serving of health. Nature's Impact, December.

Chang R, 1996. Functional properties of edible mushrooms. Nutr Rev 54(11), S91-S93

Chilton, J. 1993. What are the health benefits of mushrooms? Let's Live, Dec., pp. 24-29.

Dharmananda, S., 1988. Medicinal Mushrooms. Bestways Magazine, July, pp. 54-58.

Hobbs, C. 1997. Overcoming Chronic Fatigue (Traditional remedies for a modern desease). Veggie Life, Jan. Vol 5: #5, pp. 56-59.

Hobbs, C., 1997. Mushroom medicine. Vegetarian Times, Nov. pp. 96-98.

Hobbs, C., 1997. Medicine mushrooms. Herbs for Health, pp. 52-53. Jan/Feb.

Jones, 1997. An ancient Chinese secret promotes longevity and endurance. Healthy & Natural Journal, vol. 3, issue 3, pp. 90-93.

Jones, K., 1997. Shiitake: medicine in a mushroom. Herbs for Health. pp. 48-50, 54. Jan/Feb.

Law, David, 1996. Fungi as a platform for new medicine. Mushroom World, December, 1996.

McDougall, H. 1998. Detoxification. Veggie Life, March, pp 30 - 35.

Smith, C. 1994. Gold medal herbs. Natural Health May/June, pp. 85-87

Shirota, M, 1996. What You Should Know About Medicinal Mushrooms. Explore! vol. 7, issue 2, pp. 48-52.

Stanislaus, C., 1996. Ling zhi- medicine of kings. New Editions Health World, pp. 38-41.

Steinman, D., 1995. Potent protectors. Natural Health, Nov-Dec. pp. 92-95; 134-135.

Weil, A. 1993. Boost immunity with mushrooms. Natural Health, May-June, pp. 12-16.

Wiley, C. 1991. The medicinal side of mushrooms. Vegetarian Times, March 1991.