Immunomodulatory Properties of Plants and Mushrooms.

Plants and mushrooms are used for medicinal purposes and the screening of molecules possessing biological activities. A single plant or mushroom may produce both stimulatory and inhibitory effects on immune cells, depending on experimental conditions, but the reason behind this dichotomy remains obscure. We present here a large body of experimental data showing that water extracts of plants and mushrooms usually activate immune cells, whereas ethanol extracts inhibit immune cells. The mode of extraction of plants and mushrooms may thus determine the effects produced on immune cells, possibly due to differential solubility and potency of stimulatory and inhibitory compounds. We also examine the possibility of using such plant and mushroom extracts to treat immune system disorders.

Characterisation of a New Fungal Immunomodulatory Protein from Tiger Milk mushroom, Lignosus rhinocerotis.

Lignosus rhinocerotis (Tiger milk mushroom) is an important folk medicine for indigenous peoples in Southeast Asia. We previously reported its de novo assembled 34.3 Mb genome encoding a repertoire of proteins including a putative bioactive fungal immunomodulatory protein. Here we report the cDNA of this new member (FIP-Lrh) with a homology range of 54-64% to FIPs from other mushroom species, the closest is with FIP-glu (LZ-8) (64%) from Ganoderma lucidum. The FIP-Lrh of 112 amino acids (12.59 kDa) has a relatively hydrophobic N-terminal. Its predicted 3-dimensional model has identical folding patterns to FIP-fve and contains a partially conserved and more positively charged carbohydrates binding pocket. Docking predictions of FIP-Lrh on 14 glycans commonly found on cellular surfaces showed the best binding energy of -3.98 kcal/mol to N-acetylgalactosamine and N-acetylglucosamine. Overexpression of a 14.9 kDa soluble 6xHisFIP-Lrh was achieved in pET-28a(+)/BL21 and the purified recombinant protein was sequence verified by LC-MS/MS (QTOF) analysis. The ability to haemagglutinate both mouse and human blood at concentration ≥0.34 µM, further demonstrated its lectin nature. In addition, the cytotoxic effect of 6xHisFIP-Lrh on MCF-7, HeLa and A549 cancer cell lines was detected at IC50 of 0.34 µM, 0.58 µM and 0.60 µM, respectively.

Immunomodulatory properties of medicinal mushrooms: differential effects of water and ethanol extracts on NK cell-mediated cytotoxicity.

Medicinal mushrooms have been used for centuries in Asian countries owing to their beneficial effects on health and longevity. Previous studies have reported that a single medicinal mushroom may produce both stimulatory and inhibitory effects on immune cells, depending on conditions, but the factors responsible for this apparent dichotomy remain obscure. We show here that water and ethanol extracts of cultured mycelium from various species (Agaricus blazei Murrill, Antrodia cinnamomea, Ganoderma lucidum and Hirsutella sinensis) produce opposite effects on NK cells. Water extracts enhance NK cell cytotoxic activity against cancer cells, whereas ethanol extracts inhibit cytotoxicity. Water extracts stimulate the expression and production of cytolytic proteins (perforin and granulysin) and NKG2D/NCR cell surface receptors, and activate intracellular signaling kinases (ERK, JNK and p38). In contrast, ethanol extracts inhibit expression of cytolytic and cell surface receptors. Our results suggest that the mode of extraction of medicinal mushrooms may determine the nature of the immunomodulatory effects produced on immune cells, presumably owing to the differential solubility of stimulatory and inhibitory mediators. These findings have important implications for the preparation of medicinal mushrooms to prevent and treat human diseases.

Adaptabilidad de cepas brasileñas de Agaricus subrufescens Peck a la fructificación sobre diferentes capas de cobertura en cultivo comercial / Adaptability of Brazilian strains of Agaricus subrufescens Peck to fruiting on various casing materials in commercial crops

Antecedentes. Agaricus subrufescens Peck es un hongo cuyo cultivo ha despertado gran interés en todo el mundo en los últimos años, adquiriendo gran popularidad. Sus propiedades medicinales y culinarias hacen prever una rápida expansión del cultivo en todo el mundo. Objetivos. El trabajo plantea como objetivo la evaluación del efecto sobre los principales parámetros de producción de 3 cepas de Agaricus subrufescens que se han hecho fructificar sobre 5 capas de cobertura diferentes. Métodos. Se ha llevado a cabo un ciclo de cultivo de Agaricus subrufescens en condiciones controladas en el que se han evaluado los principales parámetros de producción. Resultados. Los mejores resultados han sido proporcionados por la cepa ABL 99/30. Las coberturas basadas en turba presentan mejor comportamiento que las basadas en suelo mineral. El mayor rendimiento (6,75 kg/m−2, eficiencia biológica 27,57 kg/dt) ha sido proporcionado por la combinación ABL 99/30-Euroveen. Conclusiones. Nuestros resultados evidencian que la combinación de la cepa ABL 99/30 utilizando una capa de cobertura basada en turba (Euroveen) ofrece un alto potencial para ser utilizada a escala comercial por el sector productor de hongos. La disponibilidad de alternativas a las especies de hongos comestibles cultivadas habitualmente puede suponer un mejor aprovechamiento de recursos y una mayor rentabilidad económica de la actividad (AU)

Background. Agaricus subrufescens Peck is a mushroom whose cultivation has aroused great interest worldwide in recent years, and is becoming increasingly popular. A rapid expansion of culture throughout the world is foreseen because of its medicinal and culinary properties. Aims. This work assesses the effect of 5 different casing layers on the production of 3 strains of Agaricus subrufescens. Methods. A growth cycle of Agaricus subrufescens under controlled conditions has been carried out. The main production parameters were evaluated. Results. The best results were provided by the ABL 99/30 strain. Peat-based casings have a better yield than those based on mineral soil. The highest yield (6.75 kg/m2, biological efficiency 27.57 kg/dt) was provided by the combination ABL 99/30-Euroveen. Conclusions. Our results suggest that the combination of the strain ABL 99/30 using a peat-based casing layer (Euroveen) offers a high potential for use on a commercial scale by the edible mushroom production sector. The availability of alternatives to the usually cultivated species can make better use of resources, and increase the profitability of this activity (AU)

Effect of immunomodulating and antiviral agent of medicinal mushrooms (immune assist 24/7) on CD4+ T-lymphocyte counts of HIV-infected patients.

Immune enhancement through the use of natural products is a potentially valuable therapeutic modality in HIV-infected people, especially those who are not good candidates for aggressive ARV therapy. One such immune enhancement, a medicinal mushroom product from the United States, is Immune Assist 24/7. In this study the effect of Immune Assist 24/7, which is a naturally derived immune-modulating and antiviral agent, on CD4+ T-lymphocyte counts was evaluated in 8 HIV-infected patients at the Sunyani Regional Hospital (Ghana). The subjects were administered three tablets of 800 mg Immune Assist 24/7 once daily (2.4 g/day) and peripheral blood samples were drawn at baseline, day 30, and day 60, and the CD4+ count measured. The study revealed that Immune Assist 24/7, used as a sole therapeutic agent without additional ARV drugs, significantly increased CD4+ T-lymphocyte populations in all of the patients. In one patient, the CD4+ T-lymphocyte count went from 4 at the baseline, to 170 cells in 60 days, representing an increase of more than 4000%. In another patient, the CD4+ count went from 88 to 470 cells within the same period. Even in the patients with the highest CD4+ counts of around 800, there was a significant elevation in the CD4+ count noted. This study did not deal with the effect of Immune Assist 24/7 on other immune parameters such as CD3+ T-lymphocyte count, natural killer cells count, or viral load among HIV-infected patients. These initial results are promising, and indicate the potential value of further evaluating the effects of Immune Assist 24/7 on other immune parameters and viral load among HIV patients, administered either as a sole therapeutic agent, as an adjuvant with standard ARV therapy, or in comparison with standard ARV therapy alone.

Preparation of Black Hoof medicinal mushroom Phellinus linteus (Berk. et M.A. Curt.) Teng (Aphyllophoromycetideae) beta-glucan sulfate and in vitro tumor cell growth inhibitory activity.

Polysaccharide beta-glucans were extracted from the medicinal mushroom Phellinus linteus (Hymenochaetaceae, Aphyllophoromycetideae) and subjected to sulfation. Chemical modification of the beta-glucan was confirmed by structural analysis, and its biological properties were compared with those of native beta-glucan. The results of Fourier transform infrared spectroscopy and elemental analysis indicated that successive preparation of the sulfated derivative yielded a degree of substitution of 0.47. Nitric oxide production measured by the bronchoalveolar lavage (BAL) experiments increased 1.5-fold after sulfation. In addition, the introduction of sulfate groups into the beta-glucan chains improved in vitro growth inhibitory activity against SNU-C2A cells. Therefore, sulfated beta-glucan extracted from Ph. linteus may be beneficial for immune support due to its incorporation of functional groups into its polymer structure.

Anticancer effects of fraction isolated from fruiting bodies of Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromycetideae): in vitro studies.

The medicinal mushroom Chaga, Inonotus obliquus (Pers.:Fr.) Pilát (Hymenochaetaceae), has been used in folk medicine in Russia, Poland, and most of the Baltic countries, as a cleansing and disinfecting measure, and as decoctions for stomach diseases, intestinal worms, liver and heart ailments, and cancer treatment. Many reports have been published concerning the health promoting functions of this mushroom, including antibacterial, hepatoprotective, anti-inflammatory, antitumor, and antioxidant activities. The purpose of the present study was evaluation of in vitro anticancer activity of fraction IO4 isolated from I. obliquus. The effect on cell proliferation, motility and viability was assessed in a range of cancer and normal cells. Chaga fraction prepared from dried fruiting bodies was subjected to anticancer evaluation in human lung carcinoma (A549), colon adenocarcinoma (HT-29), and rat glioma (C6) cell cultures. Human skin fibroblasts (HSF), bovine aorta endothelial cells (BAEC), models of rat oligodendrocytes (OLN-93), hepatocytes (Fao), rat astroglia, and mouse neurons (P19) were applied to test toxicity in normal cells. The following methods were applied: tumor cell proliferation (MTT assay and BrdU assay), cytotoxicity (LDH assay), tumor cell motility (wound assay), tumor cell morphology (May-Grünwald-Giemsa staining), and death detection (ELISA). Chaga fraction elicited anticancer effects which were attributed to decreased tumor cell proliferation, motility and morphological changes induction. Of note is the fact that it produced no or low toxicity in tested normal cells. The data presented could open interesting paths for further investigations of fraction IO4 as a potential anticancer agent.

Antitumor properties of ganoderma lucidum polysaccharidesand terpenoids: [review]

Ganoderma lucidum is an edible medicinal mushroom withimmunomodulatory and antitumor properties, which are mainly attributed to polysaccharides and triterpenesthat can be isolated from mycelia, fruiting bodies and spores. G. lucidum has been used in a powdered form, asa medicinal beverage and a nutraceutical food (usually dried). In the present review we report some historicalfacts and the experimental evidence that polysaccharides and triterpenes obtained from this mushroom presentpotential antitumor activity. Direct effects on tumor cells include induction of apoptosis and interference in thecell cycle, whereas indirect effects are based on the modulation of immune response, usually impaired bycancer cells. Data indicate that G. lucidum can be used as a complementary tool for treatment of cancerpatients.

A medicinal mushroom: Phellinus linteus.

Phellinus Linteus (Berkeley & M. A. Curtis) Teng (PL) is a medicinal mushroom that has been practiced in oriental countries for centuries to prevent ailments as diverse as gastroenteric dysfunction, diarrhea, haemorrhage and cancers. In an effort to translate the Asian traditional medicines into western-accepted therapies, scientists have demonstrated that the extracts from fruit-bodies or mycelium of PL not only stimulate the hormonal and cell-mediated immune function and quench the inflammatory reactions caused by a variety of stimuli, but also suppress the tumor growth and metastasis. Mounting evidence from different research groups has shown that PL induces apoptosis in a host of murine and human carcinomas without causing any measurable toxic effects to their normal counterparts. Recently, research has been focused on the anti-tumor effect of PL, and in particular, on its ability to enhance some conventional chemotherapeutic drugs. These studies suggest PL to be a promising candidate as an alternative anticancer agent or a synergizer for existing antitumor drugs. Hereinafter, we summarize the present progress in elucidating the mechanisms underlying the potency of PL and its anti-tumor function. The fractionation and identification of the biologically active components from PL are also briefly introduced.

Therapeutic effects for hypersensitivity pneumonitis induced by Japanese mushroom (Bunashimeji).

BACKGROUND: Bunashimeji-related hypersensitivity pneumonitis is found among workers who cultivate the mushroom in indoor facilities. An evaluation of protective measures was initiated using the outcomes of clinical, immunological, and radiological findings. METHODS: Twenty-two patients presented with symptoms of HP; all were employed cultivating Bunashimeji mushrooms in indoor facilities. After hospitalization, 6 of 22 patients quit their job to avoid exposure to spores (Avoidance group). Sixteen patients continued to work used a mask for 3 months, and were then divided into two subgroups: Mask alone (seven patients) and mask plus oral prednisolone (Mask + PSL) (nine patients). The erythrocyte sedimentation rate (ESR), serum Krebs von der Lungen-6 (KL-6), surfactant protein-D (SP-D), lymphocyte stimulation test (LST), ground-glass scores in chest high-resolution computed tomography (HRCT), and bronchoalveolar lavage (BAL) were assessed before and after treatment. RESULTS: Complete avoidance resulted in a significant decrease in LST. There was a significant decrease after PSL treatment in serum KL-6, SP-D, and total cell counts in the BAL fluid in the Mask + PSL group. In the Mask alone group, serum KL-6, SP-D, ground-glass scores in chest HRCT and total cell counts in BAL fluid showed high levels compared with the other two groups. CONCLUSIONS: Complete cessation was the best treatment for hypersensitivity pneumonitis. The use of a mask was ineffective for patients with a high serum KL-6 and SP-D concentration and severe ground-glass opacity on chest HRCT. Initial treatment with PSL is recommended for these patients with high levels of total cell counts in BAL fluid.

Successful treatment of hypersensitivity pneumonitis caused by Grifola frondosa (Maitake) mushroom using a HFA-BDP extra-fine aerosol.

We successfully treated a patient with occupational hypersensitivity pneumonitis (HP) caused by Grifola frondosa (Maitake) mushroom spore with an extra-fine aerosol corticosteroid; beclomethasone dipropionate (BDP) dissolved in hydrofluoroalkane-134a (HFA). A 49-year-old woman developed respiratory symptoms 3 months after beginning work on a mushroom farm. She was diagnosed as HP based on radiological and serological findings. Oral prednisolone therapy improved her HP and she returned to the same farm. Her HP relapsed after 5 months, and daily 400 microg of HFA-BDP was administered with gradual improvement. An extra-fine particle inhaled corticosteroid might reach appropriate alveoli to be effective therapy for mild HP.

Antibody to soluble 1,3/1,6-beta-D-glucan, SCG in sera of naive DBA/2 mice.

A branched beta-glucan from Sparassis crispa (SCG) is a major 6-branched 1,3-beta-D-glucan showing antitumor activity. In the present study, we examined the anti-SCG antibody in naive mice by ELISA. Using SCG coated plate, sera of naive DBA/1 and DBA/2 mice contained significantly higher titers of antibody than other strains of mice. Anti-SCG Ab titers of each DBA/1 and DBA/2 mice were significantly varied. Using various polysaccharide-coated plate, sera of DBA/2 mice also reacted with a beta-glucan from Candida spp. (CSBG) having 1,3-beta and 1,6-beta-glucosidic linkages. The SCG specific immunoglobulin (Ig) M but G was detected in sera. The reactivity of sera to coated SCG was neutralized by adding soluble SCG and CSBG as competitor. These results suggested that DBA/1 and DBA/2 strains carry specific and unique immunological characteristics to branched 1,3-/1,6-beta-glucan.

Treatment of cancer with mushroom products.

Cancer has been attributed to 3 causes: pollution, infection, and poor nutrition. Conventional treatments include surgery, chemotherapy, and radiotherapy. The author proposes that immunotherapy also be considered. Among other environmental influences, dietary deficiencies and carcinogenic viral infections must be investigated and treated wherever possible. It has been suggested that mushrooms, in particular, have a structure that is immunomodulatory because it resembles the proteoglycan structure in the human extracellular matrix, and both are metabolically active. Inasmuch as mitochondria have a bacterial origin, proteoglycans may have a mushroom origin. The author describes a study which shows that natural killer cells can double in number with 8 wk of treatment with Coriolus versicolor. Also described is an epidemiological survey of cancer deaths among Flammulina velutipes farmers in Japan, which found that the mushroom farmers had lower rates of cancer deaths than controls who were not involved in mushroom farming.

Purification and characterization of the allergenic components of shimeji mushroom (Tricholoma conglobatum) spore for shimeji workers' hypersensitivity pneumonitis.

Respiratory symptoms and hypersensitivity pneumonitis (HP) among mushroom workers have been well documented. Inhalation of shimeji mushroom (Tricholoma conglobatum) spore has been assumed to be the cause of HP among indoor shimeji cultivating workers. We isolated and partially characterized the allergenic components of shimeji. The sera from 9 HP patients, 10 asymptomatic shimeji workers and 15 normal individuals were examined for shimeji specific IgG and IgA antibodies by ELISA using crude shimeji extract. Shimeji specific IgG- and IgA-antibodies were higher in sera from HP patients than in sera from control subjects. Crude shimeji spore extract was separated and purified by HPLC followed by SDS-PAGE, and their antigenic activity was studied by immunoblotting with a pool of sera from patients. Sera from all HP patients showed IgG and IgA antibody activities to 21, 16 and 14 kD proteins extracted from shimeji spore. The 21 kD protein contained internal peptide amino acid sequence of Gly-Gly-Thr-Val-Ile-Asn-Leu-Leu-Gly, Gln-Arg-Phe-Glu-Glu and Gln-Gly-Ile-Tyr. These results demonstrate that shimeji spore extract contains multiple proteinous components, which have antigenic activity to react with the sera from HP patients among shimeji workers. These proteins may be the potent sensitizing allergens to cause HP among shimeji cultivating workers.

Augmentation of host's immunity by combined cryodestruction of sarcoma 180 and administration of protein-bound polysaccharide, EA6, isolated from Flammulina velutipes (Curt. ex Fr.) Sing. in ICR mice.

Augmentation of the host's immunity by combined in situ freeze-destruction of the tumor (cryosurgery) and administration of the antitumor active protein-bound polysaccharide, EA6, isolated from a hot water extract of an edible mushroom, Flammulina velutipes (Curt. ex Fr.) Sing., was investigated in sarcoma 180 bearing ICR mice. Oral administration (p.o.) of the EA6 stimulated anti-sheep red blood cells (SRBC) IgM antibody-producing activity of the spleen cells, and also delayed hypersensitivity (DTH) against SRBC in swelling of the foot pads of the tumor-bearing hosts. When EA6 p.o. was combined with cryosurgery, further augmentation of IgM-producing activity and DTH reaction to SRBC was recognized as compared with the EA6 single use. But the reticuloendothelial system of the mice, estimated by carbon clearance test, was not activated by EA6 p.o. or combined with cryosurgery.

Intensification of antitumor-immunity by protein-bound polysaccharide, EA6, derived from Flammulina velutipes (Curt. ex Fr.) Sing. combined with murine leukemia L1210 vaccine in animal experiments.

The antitumor effect of protein-bound polysaccharide, EA6, derived from fruit bodies of Flammulina velutipes (Curt. ex Fr.) Sing., when combined with a vaccine treatment was studied by the challenge test in BDF1 mice and L1210 murine leukemia system. Intensification of the antitumor effect of EA6 was dependent on doses, timing, and frequency of intraperitoneal administration of the material to the immunization by concanavalin A and/or glutaraldehyde treated L1210 vaccine. Administration of EA6 prior to the injection of the vaccine, or repeated injection of more than 4 times did not increase the life span of the animals. But when EA6 was given (40 mg/kg) after the injection of the vaccine, marked prolongation of the life span (ILS of 223%) was observed against challenging of 1 x 10(2) cells of L1210. Combined treatment of EA6 with vaccine exhibited prolonged ILS in the mice challenged with 1 x 10(3) cells of L1210. The specific immunity for L1210 induced by the vaccine was not affected by EA6.