RESUMEN
Brazil-grown outdoor-cultivated Agaricus brasiliensis KA21 fruiting body (KA21) significantly increases the production of serum anti-beta-glucan antibody. Therefore, KA21 ingestion may be useful for the prevention and alleviation of fungal infections. This study aimed to determine the effects of KA21 in fungal infections in animals. KA21 was administered to nine dogs infected with Malassezia. Notably, the anti-beta-glucan antibody titer remained unchanged or tended to decrease in the oral steroid arm, whereas in the non-steroid arm, antibody titer increased in almost all animals after KA21 ingestion. Dogs showing improved clinical symptoms exhibited increased anti-beta-glucan antibody titers. The results of this study suggest that KA21 ingestion may alleviate the symptoms of Malassezia and other fungal infections and that continuous ingestion may help prolong recurrence-free intervals. Additionally, the ingestion of KA21 during oral steroid dosage reduction or discontinuation may enable smoother steroid withdrawal.
Asunto(s)
Agaricus , Enfermedades de los Perros , Cuerpos Fructíferos de los Hongos , Malassezia , Animales , Perros , Agaricus/química , Cuerpos Fructíferos de los Hongos/química , Malassezia/efectos de los fármacos , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/tratamiento farmacológico , Dermatomicosis/veterinaria , Dermatomicosis/prevención & control , Dermatomicosis/tratamiento farmacológico , Dermatomicosis/microbiología , beta-Glucanos/administración & dosificación , beta-Glucanos/farmacología , Masculino , Brasil , Dermatitis/tratamiento farmacológico , Dermatitis/veterinaria , Dermatitis/microbiología , Dermatitis/prevención & control , Femenino , Anticuerpos Antifúngicos/sangreRESUMEN
ß-1,3-D-glucan (BG) activates innate immunity and enhances immune responses. Fungi, such as mushrooms, produce a relatively large amount of BG, the structure and molecular weight of which varies depending on the species of fungi. This study was conducted to develop a detection probe for quantifying or detecting BG from fungi using BG-binding proteins. The binding properties of a new ß-glucan recognition protein (BGRP) against various BGs were compared. With reference to the amino acid sequences of BGRP in insects, an artificial BGRP (supBGRP) was designed with higher production efficiency using gene recombination technology. SupBGRP was produced in Escherichia coli with high efficiency, and its reactivity with BG from fungi was the highest among the BG-binding proteins examined. SupBGRP exhibited high reactivity with 1,6-branched BG and will be useful for the quantification and detection of fungal BG.
Asunto(s)
Agaricales/química , beta-Glucanos/aislamiento & purificación , beta-Glucanos/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , beta-Glucanos/químicaRESUMEN
The edible mushroom Agaricus brasiliensis contains a large amount ß-glucan, which is mainly composed of a ß-1,6-glucan structure. In this study, we investigated the effect of A. brasiliensis strain KA21 on the anti-ß-glucan antibody titer in healthy humans and the role of antibodies as an immunomodulator. Twenty-two healthy volunteers were fed the dried fruiting body of A. brasiliensis (900 or 1500 mg/day) for 12 weeks. The anti-ß-glucan antibody titer in the serum was determined by enzyme-linked immunosorbent assay. Immunoglobulin G (IgG) against ß-glucan was significantly upregulated after intake of A. brasiliensis. Murine experiments demonstrated improvement of anti-ß-glucan antibody production after intraperitoneal injection of Agaricus-derived ß-glucan. To understand the role of antibody against ß-glucan in exclusion of pathogenic fungi, we examined the interaction between HL-60 cells and antibody-treated heat-killed Candida albicans. Flow cytometry analysis indicated the upregulation of Candida-positive HL-60 cells after treatment with human IgG, whereas the competitive assay demonstrated that the main epitope of Candida-reacted IgG was the ß-1,6-glucan structure. Binding between HL-60 and IgG-opsonized C. albicans was suppressed by anti-Fcγ receptor 1 (FcγRI) neutralizing antibody. Finally, using FcγRI-expressed cells with the nuclear factor of activated T-cell reporter assay, we demonstrated that higher titers of anti-ß-glucan IgG can induce stronger Fc receptor-mediated cell activation through the formation of an antibody-ß-glucan complex. In conclusion, oral ingestion of A. brasiliensis KA21 promotes anti-ß-glucan antibody production and may contribute to preventing fungal infection through the activation of immune cells by forming antibody-ß-glucan complexes via an FcγR-dependent pathway.
Asunto(s)
Agaricus , Animales , Formación de Anticuerpos , Glucanos , Voluntarios Sanos , Humanos , Ratones , beta-GlucanosRESUMEN
The royal sun medicinal mushroom, Agaricus brasiliensis, is a health food material that helps to improve quality of life. A. brasiliensis has long been used as a tea by extraction with cold and hot water. Our group has been investigating the immunopharmacological activities of the A. brasiliensis KA21 strain, which is cultivated outdoors. We prepared cold water (AgCWE) and hot water (AgHWE) extracts of this strain. AgCWE contained a larger proportion of proteins, including enzymes, and showed a brownish color during the extraction process. By contrast, chemical and immunochemical analyses revealed that AgHWE contained large amounts of ß-1,3-/1,6-glucans. In an attempt to elucidate the immunochemical characteristics of AgCWE, reactivities to immunoglobulin (Ig) preparations for intravenous injection were analyzed and compared with standard materials. To characterize brownish high-molecular weight components, standard phenol compounds such as caffeic acid (CA), trans-ferulic acid (FA), and coumaric acid (CouA) were polymerized to brownish polymerized polyphenols (PPPs) (i.e., polymerized CA, polymerized FA, and polymerized CouA) by laccase or peroxidase. The results obtained revealed that intravenous Ig reacted with all PPPs and PPPs cross-reacted with AgCWE and AgHWE. The isotype of the anti-PPP antibody was found to be IgG1, in contrast to that of the ß-glucan antibody, which was mainly IgG2. These results strongly suggest that A. brasiliensis extracts contain immunoreactive components against various classes of Igs.
Asunto(s)
Agaricus/inmunología , Anticuerpos Antifúngicos/inmunología , Mezclas Complejas/inmunología , Alimentos , Humanos , Inyecciones Intravenosas , Lignanos/inmunología , Agua , beta-Glucanos/inmunologíaRESUMEN
ãAgaricus is known to have immunostimulatory and anti-tumor effects. However, the antiviral effects of Agaricus have not yet been examined. In the present study, the antiviral effects of an extract of Agaricus brasiliensis KA21 (AE) on the H1N1 influenza virus (PR8 strain) were investigated.ãThe anti-influenza virus effects of AE were examined by using the plaque formation inhibition test. AE inhibited the plaque formation of PR8 in a dose-dependent manner: 98 and 50% (IC50) inhibition at 2.5 and 0.99 mg/mL, respectively. To elucidate the mechanisms of AE, the direct actions and adsorption and invasion inhibition of AE were examined, and were found to have no inhibitory effect on PR8 infection. Thus, in vitro antiviral effects may somehow inhibit PR8 after the viral invasion of cells. These results demonstrated that it is expected that AE can effectively prevent the spread of the influenza virus.
Asunto(s)
Agaricus/química , Antivirales/farmacología , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Línea Celular , Relación Dosis-Respuesta a Droga , Humanos , Gripe Humana/tratamiento farmacológico , Concentración 50 Inhibidora , Replicación ViralRESUMEN
We conducted an open-label study in which food containing Agaricus brasiliensis KA21 was consumed continuously for 12 weeks. A questionnaire for subjective evaluation of the efficacy of this food (hereafter, subjective evaluation questionnaire) revealed significant improvements compared with before its intake; there were improvements in the scores of the amounts of hair loss and gray hair, fatigue and general malaise, eye strain, shoulder stiffness, coldness of extremities, difficulty staying awake during the day, and ease of getting out of bed. These findings suggest that intake of food containing A. brasiliensis KA21 results in the above-mentioned subjectively evaluated improvements, and the possibility that A. brasiliensis KA21 improves the body's immunity. Moreover, no issues regarding the safety of the test food were found.
Asunto(s)
Agaricus , Dieta , Calidad de Vida , Actividades Cotidianas , Adulto , Ojo , Fatiga/dietoterapia , Femenino , Cabello , Voluntarios Sanos , Humanos , Inmunidad , Masculino , Persona de Mediana Edad , Rango del Movimiento Articular , Autoinforme , Hombro , Sueño , Encuestas y Cuestionarios , VerdurasRESUMEN
BACKGROUND: Agaricus brasiliensis (A. brasiliensis) is a medicinal mushroom that exerts various pharmacological actions. We previously demonstrated that different cultivation conditions altered the activity of the polyphenol-related enzymes from this mushroom. However, the influence of cultivation conditions on the antioxidant activity of the fruiting bodies remains unclear. Therefore, in this study we compared the antioxidative effects of fruiting bodies of A. brasiliensis cultivated outdoors and indoors. In addition, we assessed whether different cultivation methods affected the hepatoprotective effects against CCl4-induced liver injury. METHODS: We assessed the antioxidative effects of mushrooms cultivated in open-air or indoors using the DPPH radical-scavenging assay. Furthermore, we prepared experimental feeds containing outdoor- or indoor-cultivated A. brasiliensis. Acute liver injury was induced by CCl4 injection in mice that consumed feed containing outdoor- or indoor-cultivated A. brasiliensis. The hepatoprotective effects of these mushrooms were then evaluated by monitoring the reduction in the circulating levels of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase. The significance of the differences between the means was assessed using Student's t-test. Finally, histopathological analysis of liver was performed. RESULTS: In the DPPH assay, the antioxidant activity of outdoor-cultivated A. brasiliensis was higher than that of indoor-cultivated mushroom. Moreover, in the mouse model of CCl4-induced hepatitis, the oral administration of outdoor-cultivated A. brasiliensis reduced liver damage significantly, but indoor-cultivated mushrooms failed to inhibit hepatitis. The hepatoprotective effects of outdoor-cultivated A. brasiliensis were observed even when ingestion commenced only 1 day before CCl4 injection, and these effects were not affected by excessive heat treatment. CONCLUSIONS: Outdoor cultivation significantly enhanced the antioxidative activity of A. brasiliensis fruiting bodies. In addition, outdoor-cultivated A. brasiliensis was more effective at protecting against CCl4-induced liver injury in mice than mushrooms grown in a greenhouse.
Asunto(s)
Agaricus/química , Agricultura/métodos , Antioxidantes/uso terapéutico , Productos Biológicos/uso terapéutico , Enfermedad Hepática Inducida por Sustancias y Drogas/tratamiento farmacológico , Hígado/efectos de los fármacos , Alanina Transaminasa/sangre , Animales , Antioxidantes/farmacología , Aspartato Aminotransferasas/sangre , Productos Biológicos/farmacología , Compuestos de Bifenilo/metabolismo , Tetracloruro de Carbono , Enfermedad Hepática Inducida por Sustancias y Drogas/sangre , Cuerpos Fructíferos de los Hongos , Hepatitis/sangre , Hepatitis/tratamiento farmacológico , L-Lactato Deshidrogenasa/sangre , Hígado/enzimología , Masculino , Ratones Endogámicos ICR , Oxidación-Reducción , Picratos/metabolismo , Polifenoles/farmacología , Sustancias Protectoras/farmacología , Sustancias Protectoras/uso terapéuticoRESUMEN
Laccase isozymes have been identified in several fungi. We report the cloning of 4 laccase genes from the medicinal mushroom Agaricus brasiliensis. The lac1 gene contained a 1560-base pair (bp) open reading frame (ORF) encoding 520 amino acids that was interrupted with 14 introns in genomic DNA. The deduced amino acid sequence indicated a multicopper oxidase signature 1 and 2 multicopper oxidase signature 2. The lac2 gene contained a 1566-bp ORF encoding 522 amino acids that was interrupted with 13 introns in genomic DNA. A number of different nucleotides were observed in whole regions containing the substitution of amino acid residues (lac2a and lac2b). The partial DNA fragments of lac3 and lac4 genes were subcloned using the semi-random two-step polymerase chain reaction method. The lac3 and lac4 genes contained coding sequences with a 1575-bp ORF encoding 525 amino acids and a 1584-bp ORF encoding 528 amino acids, respectively. However, the whole complementary DNA fragment of both laccases could not be amplified with polymerase chain reaction against the complementary DNA library; therefore, introns were deduced based on the GT-AG rule and multiple alignment of laccases from other fungi, which showed high identity. All laccases from A. brasiliensis conserved the fungal laccase signature sequence and suggest 2 subfamilies according to the location of introns and phylogenetic analysis. The genes lac2 and lac4 had a high degree of identity, and the lac2a gene was located upstream of the lac4 gene.
Asunto(s)
Agaricus/enzimología , Agaricus/genética , Lacasa/genética , Secuencia de Aminoácidos , Secuencia de Bases , Basidiomycota , Clonación Molecular , Análisis por Conglomerados , ADN de Hongos/química , ADN de Hongos/genética , Hongos , Intrones , Lacasa/química , Modelos Moleculares , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Filogenia , Reacción en Cadena de la Polimerasa , Conformación Proteica , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de SecuenciaRESUMEN
The present study aimed to clarify the effects of Agaricus brasiliensis KA21 (i.e., Agaricus blazei) mushroom on circulatory function. Spontaneously hypertensive rats (SHRs) were fed 10% A. blazei-containing pellets (agaricus group) or normal pellets (control group) for 5 weeks from 6 to 11 weeks of age. For Experiment 1, tail blood pressure and heart rate were measured in the conscious SHRs. For Experiment 2, echocardiographic and blood biochemical measurements were performed in the anesthetized SHRs. In Experiment 1, blood pressure and heart rate were significantly lower in the agaricus group compared with the control group throughout the observation period. In Experiment 2, the agaricus group also showed a significant decrease in cardiac output accompanied by a decrease in heart rate and an increase in early and late ventricular filling velocity (E/A ratio). Moreover, levels of escape enzymes such as creatine kinase (CK), CK-BB, CK-MB, asparate aminotransferase, lactate dehydrogenase, and aldolase were significantly lower than in the control group. We concluded that the ingestion of feed containing A. brasiliensis KA21 can improve hypertensive cardiovascular hemodynamics by decreasing the working load of the heart, presumably by lowering the sympathetic nervous tone in SHRs.
Asunto(s)
Agaricus/metabolismo , Antihipertensivos/metabolismo , Circulación Sanguínea , Hipertensión/dietoterapia , Animales , Frecuencia Cardíaca , Humanos , Hipertensión/fisiopatología , Ratas , Ratas Endogámicas SHRRESUMEN
Lignin-like polymerized polyphenols strongly activate lymphocytes and induce cytokine synthesis. We aimed to characterise the mechanisms of action of polymerized polyphenols on immunomodulating functions. We compared the reactivity of leukocytes from various organs to that of polymerized polyphenols. Splenocytes and resident peritoneal cavity cells (PCCs) responded to polymerized polyphenols and released several cytokines, whereas thymocytes and bone-marrow cells showed no response. Next, we eliminated antigen-presenting cells (APCs) from splenocytes to study their involvement in cytokine synthesis. We found that APC-negative splenocytes showed significantly reduced cytokine production induced by polymerized polyphenols. Additionally, adequate interferon-γ (IFN-γ) induction by polymerized polyphenols was mediated by the coexistence of APCs and T cells because the addition of T cells to PCCs increased IFN-γ production. Furthermore, inhibition of the T cell-APC interaction using neutralising antibodies significantly decreased cytokine production. Thus, cytokine induction by polymerized polyphenols was mediated by the interaction between APCs and T cells.
Asunto(s)
Células Presentadoras de Antígenos/inmunología , Comunicación Celular/efectos de los fármacos , Factores Inmunológicos/farmacología , Interferón gamma/biosíntesis , Polifenoles/farmacología , Animales , Células Presentadoras de Antígenos/citología , Células Presentadoras de Antígenos/efectos de los fármacos , Células Cultivadas , Factores Inmunológicos/química , Leucocitos/citología , Leucocitos/efectos de los fármacos , Leucocitos/inmunología , Lignina , Ratones , Ratones Endogámicos C57BL , Polimerizacion , Polifenoles/químicaRESUMEN
The Royal Sun medicinal mushroom, Agaricus brasiliensis, is used as a natural health product. In Japan, however, the quality control of some of these mushroom products has been viewed as a safety problem. Focusing on the quality control of A. brasiliensis KA21, we have performed several safety studies. To date, we have established evidence that this mushroom can be used safely as an immunostimulant and to mediate biochemical parameters associated with obesity or diabetes. Furthermore, to improve the manufacturing process of this mushroom, we have studied the relationship between its pharmaceutical actions and the conditions of its cultivation and thermal management. The purpose of this review is to report the findings of basic and clinical studies of the fruit body of A. brasiliensis KA21.
Asunto(s)
Agaricus/química , Alimentos Funcionales , HumanosRESUMEN
The aim of this work was to characterize an antibody response to ß-glucan (BG), a major component of the fungal cell wall, at each isotype in human sera. The titer and reactivity of the anti-human BG antibody were examined using enzyme-linked immunosorbant assay plates coated with Candida soluble cell wall ß-glucan as a standard antigen. The antibody was detected using anti-human immunoglobulin (Ig) G, IgM, and IgA. Its major class was IgG in all subjects. The antibody titer varied significantly. The anti-BG antibody showed greater reactivity to ß-glucans derived from pathogenic fungi than monoglycosyl-branched ß-glucans derived from mycelia culture medium. In addition, it was suggested that the anti-BG IgM antibody was bound relatively strongly to the ß1,3-glucan backbone and the anti-BG IgG antibody to ß1,6-glucan. The anti-BG antibody plays a variety of roles, according to class, in the host's response to fungi. We propose a new index of human response to ß-glucan that effects the understanding of the response to ß-glucan in humans.
Asunto(s)
Anticuerpos/sangre , Aspergillus/citología , Pared Celular/química , Pared Celular/inmunología , beta-Glucanos/inmunología , Adulto , Especificidad de Anticuerpos , Aspergillus/metabolismo , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
Agaricus brasiliensis is a well-known medicinal mushroom. We have previously demonstrated that Agaricus-derived polysaccharides exhibit potent antitumor effects; however, the underlying mechanism(s) have not been elucidated yet. In this study, we examined the immunoenhancing activities of Agaricus extracts. Agaricus-derived polysaccharides were characterized as 1,6-ß-glucan with a small amount of 1,3-ß-glucan using anti-ß-glucan antibody and nuclear magnetic resonance analysis. These polysaccharides strongly induced the production of various cytokines from both murine splenocytes and bone marrow-derived dendritic cells in the presence of exogenous granulocyte-macrophage colony-stimulating factor. Polysaccharide-induced cytokine production was significantly reduced in bone marrow-derived dendritic cells derived from dectin-1-deficient mice. Furthermore, a binding assay revealed that the Agaricus-derived polysaccharides can be recognized by dectin-1, a pivotal receptor for 1,3-ß-glucan. Taken together, our results clearly indicate that the immunostimulation induced by Agaricus-derived polysaccharides is exerted, at least in part, via dectin-1 in combination with granulocyte-macrophage colony-stimulating factor.
Asunto(s)
Agaricus , Factores Inmunológicos/farmacología , Lectinas Tipo C/inmunología , beta-Glucanos/farmacología , Animales , Células de la Médula Ósea/citología , Células Cultivadas , Citocinas/inmunología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Células HEK293 , Humanos , Lectinas Tipo C/deficiencia , Lectinas Tipo C/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Bazo/citologíaRESUMEN
As part of the safety evaluation of culinary-medicinal Royal Sun Agaricus, Agaricus brasiliensis KA21, for human consumption, we performed the bacterial reverse mutation test, the mouse micronucleus test, and mouse lymphoma test using A. brasiliensis KA21 as the test substance. The reverse mutation test utilized five bacterial strains, including Salmonella typhimurium TA100, TA1535, TA98, and TA1537, and Escherichia coli WP2 uvrAO. For the micronucleus test we used mice. For the mouse lymphoma test, we used one of the most commonly used mammalian cell mutagenesis systems; the L5178YTK +/- mouse lymphoma-TK assay detects the mutations at the thymidine kinase locus caused by base-pair changes, frameshift, and small deletions. All the tests were conducted according to the guidelines for genotoxicity testing of drugs by the Ministry of Health, Labor, and Welfare, Japan. In the bacterial reverse mutation test, no toxicity was observed up to a dose of 5,000 µg/plate. In the mouse micronucleus test, no toxicity was noted up to a dose of 1 g/kg body weight. In the mouse lymphoma test, frequency of the mutation was equal both in the presence or absence of KA21. Supporting the long history of human consumption of A. brasiliensis, the data shown in this study strongly indicate the safety of this mushroom.
Asunto(s)
Agaricus/química , Alimentos/efectos adversos , Animales , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Inocuidad de los Alimentos , Humanos , Linfoma , Masculino , Ratones , Pruebas de Mutagenicidad , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genéticaRESUMEN
High-molecular weight polymerised polyphenols have been shown to exhibit anti-influenza virus, anti-HIV, and anti-cancer activities. The purpose of this study was to evaluate the immunomodulating activities of enzymatically polymerised polyphenols, and to clarify the underlying mechanisms of their effects. The cytokine-inducing activity of the enzymatically polymerised polyphenols derived from caffeic acid (CA), ferulic acid (FA), and p-coumaric acid (CoA) was investigated using murine splenocytes. Polymerised polyphenols, but not non-polymerised polyphenols, induced cytokine synthesis in murine splenocytes. Polymerised polyphenols induced several cytokines in murine splenocytes, with interferon-γ (IFN-γ) and granulocyte-macrophage colony-stimulating factor (GM-CSF) being the most prominent. The underlying mechanisms of the effects of the polymerised polyphenols were then studied using neutralising antibodies and fluorescent-activated cell sorting (FACS) analysis. Our results show that polymerised polyphenols increased IFN-γ and GM-CSF production in splenocytes. In addition, the anti-CD4 neutralised monoclonal antibody (mAb) inhibited polymerised polyphenol-induced IFN-γ and GM-CSF secretion. Moreover, polymerised polyphenols bound directly to a recombinant CD4 protein, and FACS analysis confirmed that interaction occurs between polymerised polyphenols and CD4 molecules expressed on the cell surface. In this study, we clearly demonstrated that enzymatic polymerisation confers immunoactivating potential to phenylpropanoic acids, and CD4 plays a key role in their cytokine-inducing activity.
Asunto(s)
Antígenos CD4/metabolismo , Citocinas/metabolismo , Factores Inmunológicos/farmacología , Polifenoles/farmacología , Bazo/metabolismo , Animales , Anticuerpos Monoclonales/inmunología , Antígenos CD4/genética , Antígenos CD4/inmunología , Células Cultivadas , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Factores Inmunológicos/química , Interferón gamma/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Polimerizacion , Polifenoles/química , Unión Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Bazo/citología , Bazo/efectos de los fármacosRESUMEN
Agaricus brasiliensis (Agaricus blazei Murrill) is well known as a medicinal mushroom. Fruit body of A. brasiliensis is rich in ß-glucan and has shown benefits for various diseases. Both hot and cold water extraction are traditional methods for intake of this mushroom extract. In the present study, we prepared cold water extract of the fruit body of A. brasiliensis (ACWS). The 1,3-ß-glucan segment of this fraction was too small and did not interact with the 1,3-ß-glucan receptor, dectin-1. However, ACWS could induce production of various cytokines including IL-6 from murine splenocytes. Therefore, we aimed to identify the receptor that modulates IL-6 production using ACWS. We focused our attention on Toll-like receptors (TLRs) and examined them as follows. (i) The interaction between TLRs and ACWS was screened using HEK293 cells transfected with TLR plasmid. (ii) IL-6 production from splenocytes induced by ACWS was inhibited by treatment of anti-TLR antibodies. (iii) Direct binding activity between TLR protein and ACWS was assessed by ELISA-like assay. ACWS was found to activate HEK293 cells via TLR2, 4 and 5. However, only anti-TLR2 monoclonal antibody suppressed IL-6 production from splenocytes. In addition, ACWS has the ability to bind directly to TLR2 protein. Accordingly, we suggest that fruit body of A. brasiliensis has some water-soluble TLR ligand complexes, and TLR2 on splenocytes strongly induces IL-6 production.
Asunto(s)
Agaricus/química , Mezclas Complejas , Interleucina-6/biosíntesis , Receptor Toll-Like 2/metabolismo , beta-Glucanos , Animales , Mezclas Complejas/química , Mezclas Complejas/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Células HEK293 , Humanos , Interleucina-6/genética , Lectinas Tipo C/agonistas , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Masculino , Ratones , Receptor Toll-Like 2/genética , beta-Glucanos/química , beta-Glucanos/farmacologíaRESUMEN
The Royal Sun mushroom, the Himematsutake culinary-medicinal mushroom, Agaricus brasiliensis has several polyphenoloxidase activities in a broad sense. Here we report the partial purification of tyrosinase-type polyphenoloxidase (PPO). PPO is purified from A. brasiliensis without browning using a two-phase partitioning with Triton X-114 and ammonium sulfate fractionation. Partially denaturing SDS-PAGE (sodium dodecyl sulfate-polyacrylamide electrophoresis) staining with L-3,4-dihydroxyphenylalanine was performed and the indicated molecular sizes were approximately 70 kDa and 45 kDa. The purified enzyme is in its latent state and can be activated maximally in the presence of 1.6 mM sodium dodecyl sulfate (SDS). This enzyme catalyzes two distinct reactions, monophenolase and diphenolase activity, and the monophenolase activity showed a lag time typical of polyphenoloxidase. The K(m) value for 4-tert-butylcatechol was quite similar in the presence and absence of SDS, but the apparent V(max) value was increased 2.0-fold by SDS. Mimosine was a typical competitive inhibitor with K(i) values of 138.2 microM and 281.0 microM n the presence and absence of SDS, respectively.
Asunto(s)
Agaricus/enzimología , Catecol Oxidasa/metabolismo , Cuerpos Fructíferos de los Hongos/enzimología , Catecol Oxidasa/antagonistas & inhibidores , Catecol Oxidasa/aislamiento & purificación , Electroforesis en Gel de Poliacrilamida , Activación Enzimática/efectos de los fármacos , Proteínas Fúngicas/aislamiento & purificación , Proteínas Fúngicas/metabolismo , Concentración de Iones de Hidrógeno , Cinética , Monofenol Monooxigenasa/aislamiento & purificación , Monofenol Monooxigenasa/metabolismo , Dodecil Sulfato de Sodio/farmacología , TemperaturaRESUMEN
ß-1,3-glucan is a major component of fungal cell walls with various biological activities, including effects on the production of inflammatory mediators in vivo and in vitro. However, few reports have examined its influence on the fungal cell itself. In this study, the influences of ß-1,3-glucan on the growth and cell wall structure of fungi was examined. Aspergillus fumigatus was cultured with a synthetic medium, C-limiting medium, in the presence or absence of ß-1,3-glucan. Hyphal growth was promoted in liquid and solid-cultures by adding ß-1,3-glucan. Glucose and dextran did not induce growth. The influence on cell wall structure of the ß-glucan-added cultures was examined by enzymolysis and NMR spectroscopy and the amount of ß-1,3-glucan found to be changed. ß-1,3-glucan has been widely detected in the environment. In this study, it was demonstrated that ß-1,3-glucan causes promotion of the growth, and a change in the cell wall architecture, of Aspergillus. Unregulated distribution of ß-1,3-glucan would be strongly related to the incidence of infectious diseases and allergy caused by Aspergillus spp.
Asunto(s)
Aspergillus/efectos de los fármacos , Pared Celular/efectos de los fármacos , beta-Glucanos/farmacología , Aspergillus/crecimiento & desarrollo , Pared Celular/química , Medios de Cultivo , beta-Glucanos/metabolismoRESUMEN
beta-Glucan is a major component of the cell walls and pathogen-associated microbial patterns of fungi. We previously reported the presence of an antibody which reacts to beta-glucan, anti-beta-glucan (BG) antibody, in human sera. In livestock and domestic pets, the antibody's response to fungal cell wall beta-glucan is little understood. In this study, we examined the existence and reactivity of anti-BG antibody in various animal species. We demonstrated the presence of the anti-BG antibody in each animal's serum. Individual differences in the titer existed. The antibody was highly reactive to Candida solubilized cell wall beta-glucan (CSBG) while reacting little to grifolan (GRN) from Grifola frondosa. This suggested that the anti-BG antibody interacted with fungal cell wall beta-glucan and participated in the immune-response to pathogenic fungi.
Asunto(s)
Anticuerpos Antifúngicos/sangre , Pared Celular/inmunología , Hongos/citología , Hongos/inmunología , beta-Glucanos/inmunología , Animales , Especificidad de Anticuerpos , Bovinos , Pollos , Perros , Ensayo de Inmunoadsorción Enzimática , Cabras , Cobayas , Haplorrinos , Caballos , Inmunidad Humoral , Micosis/inmunología , Micosis/microbiología , Conejos , Porcinos , TurquíaRESUMEN
We performed studies on murine models and human volunteers to examine the immunoenhancing effects of the naturally outdoor-cultivated fruit body of Agaricus brasiliensis KA21 (i.e. Agaricus blazei). Antitumor, leukocyte-enhancing, hepatopathy-alleviating and endotoxin shock-alleviating effects were found in mice. In the human study, percentage body fat, percentage visceral fat, blood cholesterol level and blood glucose level were decreased, and natural killer cell activity was increased. Taken together, the results strongly suggest that the A. brasiliensis fruit body is useful as a health-promoting food.