Immunomodulation with ß-glucan from Pleurotus ostreatus in patients with endocrine-dependent breast cancer.

Aim: To evaluate the effect of pleuran (ß-glucan from Pleurotus ostreatus) administration on the immune profile of patients with endocrine-dependent breast cancer (clinical stages I-II) in clinical and imaging remission. Methodology: Antitumor cellular immunity (CD19+, CD3+, CD4+ and CD8+ T lymphocytes and natural killer cells) of 195 patients (49 in the pleuran group and 146 in the control group) was measured by flow cytometry. Results: We observed a significant increase in the absolute number of CD3+, CD19+, CD4+ and CD8+ T lymphocytes in the pleuran group compared with the control group. Conclusion: Our results suggest potential benefit of continuous pleuran administration on immune rehabilitation of cellular antitumor immunity and better prognosis in breast cancer patients in remission.

Lay abstract We aimed to evaluate the effect of pleuran (ß-glucan from oyster mushroom) on the selected immune parameters of patients with breast cancer in remission. We studied antitumor cellular immune parameters of 195 patients (49 in the pleuran group and 146 in the control group) by means of flow cytometry. After 12 months, we measured a significant increase of cytotoxic T lymphocytes in the pleuran group compared with a significant decrease in the control group. Our results suggest potential benefit of long-term administration of pleuran on antitumor cellular immunity and better prognosis in breast cancer patients in remission.

Increased Response of Human T-Lymphocytes by Dendritic Cells Pulsed with HPV16E7 and Pleurotus sajor-caju-ß-glucan (PBG).

BACKGROUND: Infection with human papillomavirus type 16 (HPV-16) is known to cause cervical cancer, hence the several HPV therapeutic vaccines are developed in E7 oncoproteins and targeted on cell-mediated immunity. Human dendritic cells (HuDCs) are extensively employed in HPV therapeutic vaccines as the carrier or platform for inducing adaptive immune responses. However, the immunomodulators need to be further investigated for vaccine effects. Gray oyster mushroom (Pleurotus sajor-caju) containing ß-glucans is a potent immunomodulator with potential to be used in vaccines. OBJECTIVE: To study the effect of Pleurotus sajor-caju-ß-glucan Polysaccharides (PBG) on human T-lymphocytes by use of the HuDCs' antigen presentation platform for HPV16 vaccine. METHODS: The HPV16-E7 recombinant proteins were constructed in E. Coli. HuDCs pulsed with E7 peptide were cocultured with the T-lymphocytes treated with and without PBG. The number of T-lymphocytes(CD4; CD8) was detected by flowcytometry, and the viral response of T-lymphocytes was measured via IFN-γ release. RESULTS: The PBG treated group of T-lymphocytes cocultured with the HuDCs pulsed by the HPV16-E7 proteins showed significantly higher numbers of T-lymphocytes and IFN-γ release compared with T-lymphocytes without PBG in vitro. Moreover, a significant improvement in the level of specific IgG neutralizing antibodies to HPV was found in a murine model. Further observed was an increase in the expansion of helper and cytotoxic T-cells and IFN-γ releases in human system. CONCLUSION: PBG treatment of T-lymphocytes could be a useful option for prophylactic and therapeutic vaccines in cervical cancer.

Activity-guided isolation and structural identification of immunomodulating substances from Pleurotus eryngii byproducts.

A two-step extraction process was used to investigate the bioactive substances from the byproducts of the fungus Pleurotus eryngii. Methanol extraction followed by extraction using different polar solvents coupled with silica column chromatography was carried out to separate the bioactive components. The immunomodulating activity of the extracts was investigated using Ana-1 mouse macrophage cells. At 25µg/mL, the butyl alcohol extract showed the highest enhancement of Ana-1 cell proliferative activity and secretion of cytokines. Using silica-gel column chromatography, the butyl alcohol extract was purified to seven elution fractions. Fr. 2 showed the highest promotion of Ana-1 macrophage cell proliferation and TNF-α, IL-6 secretion. HPLC-QTOF-MS/MS was used to determine the structure of the active substance in Fr. 2. Adenosine was identified as the primary active component in Fr. 2 with a chemical formula of C10H13N5O4. Adenosine may be useful as a natural immunostimulatory agent and anti-tumor agent.

A recombinant fungal compound induces anti-proliferative and pro-apoptotic effects on colon cancer cells.

Finding intracellular pathways and molecules that can prevent the proliferation of colon cancer cells can provide significant bases for developing treatments for this disease. Ostreolysin (Oly) is a protein found in the mushroom Pleurotus ostreatus, and we have produced a recombinant version of this protein (rOly).We measured the viability of several colon cancer cells treated with rOly. Xenografts and syngeneic colon cancer cells were injected into in vivo mouse models, which were then treated with this recombinant protein.rOly treatment induced a significant reduction in viability of human and mouse colon cancer cells. In contrast, there was no reduction in the viability of normal epithelial cells from the small intestine. In the search for cellular targets of rOly, we showed that it enhances the anti-proliferative activity of drugs targeting cellular tubulin. This was accompanied by a reduction in the weight and volume of tumours in mice injected with rOly as compared to their respective control mice in two in vivo models.Our results advance the functional understanding of rOly as a potential anti-cancer treatment associated with pro-apoptotic activities preferentially targeting colon cancer cells.

The combination of Pleurotus ferulae water extract and CpG-ODN enhances the immune responses and antitumor efficacy of HPV peptides pulsed dendritic cell-based vaccine.

Our previous study reported that the combination of Pleurotus ferulae water extract (PFWE) and CpG (PFWE+CpG) enhanced the maturation and function of dendritic cells (DCs). Here, we investigated the effects of PFWE+CpG on the immune responses and antitumor efficacy of DC-based vaccine. We observed that all of HPV E6 and E7 peptides pulsed DCs (HPV-immature DCs, HPV+PFWE-, +CpG- or +PFWE+CpG-DCs) induced antigen-specific CD8(+) T cell responses and HPV+PFWE+CpG-DCs induced highest level of CD8(+) T cell responses. The antitumor efficacy of HPV-DCs vaccines was evaluated in TC-1 tumor mouse model. The early therapeutic study showed that HPV+PFWE-, +CpG- and +PFWE+CpG-DCs greatly inhibited tumor growth. Moreover, HPV+PFWE+CpG-DCs controlled tumor growth at a faster rate compared to other groups. These three groups induced HPV-specific CD8(+) T cell responses and significantly decreased the frequencies of induced regulatory T cells (iTregs: CD4(+)CD25(-)Fopx3(+)). However, only HPV+PFWE+CpG-DCs significantly decreased the frequency of natural Tregs (nTregs: CD4(+)CD25(+)Fopx3(+)). Furthermore, HPV+PFWE+CpG-DCs also significantly inhibited tumor growth in the late therapeutic study. The results showed that PFWE+CpG enhanced the immune responses and antitumor efficacy of DC-based vaccine, suggesting that PFWE+CpG might be the potential candidate for the generation of clinical-grade mature DCs.

Occupational asthma in a fruit and vegetables vendor.

BACKGROUND: This study presents a case report of a woman running a fruit and vegetables market stall, occupationally exposed to mushroom spores, who developed asthmatic symptoms. Basidiospores are allergens that can give rise to sensitization and subsequently trigger an allergic reaction, such as contact dermatitis, rhino-conjunctivitis, asthma and hypersensitivity pneumonitis. METHODS: A 40-year-old atopic woman, who had worked at a market stall selling fruit and vegetables for the past 26 years in a small town market in southern Italy, came to our attention at the Occupational Medicine Unit, complaining of a 3-year history of dyspnoea, cough, and wheezing during autumn and winter. RESULTS: Spirometry showed a severe mixed type respiratory deficit with a significant bronchodilator response obtained with salbutamol administration. Skin prick tests to common aeroallergens were negative. Prick-to-prick test with fresh Pleurotus ostreatus was positive. We evaluated the size of the skin wheals and compared them to the positive control according to the Skin Index. An on-the-spot inspection of her store was performed and environmental conditions were identified that did not allow a satisfactory air exchange. CONCLUSIONS: The clinical history and the results of the allergologic investigations, plus an on-the-spot inspection, permitted us to ascertain that this subject had developed occupational asthma due to Pleurotus ostreatus spores. We advised her to stop handling and selling mushrooms, and she has no longer suffered asthmatic symptoms, over seven years of follow-up, and regularly continued to work without needing to take anti-asthma drugs.

Induction of a T-Helper 1 (Th1) immune response in mice by an extract from the Pleurotus eryngii (Eringi) mushroom.

To assess the effect of edible mushroom extracts on the induction of T-helper 1 (Th1) immunity, we examined differences in interferon-gamma (IFN-γ) and interleukin (IL)-4 production in mice induced by hot-water extracts of 15 species of edible mushroom. Extracts from Agaricus bisporus, Flammulina velutipes, Hypsizigus marmoreus, Lentinula edodes, and Lyophyllum decastes induced both IFN-γ and IL-4 production in mice, whereas extracts from Pleurotus ostreatus only induced IL-4. In contrast, extracts from Agaricus blazei, Grifola frondosa, Morchella esculenta, Pholiota nameko, Pleurotus citrinopileatus, and Pleurotus eryngii induced only IFN-γ production. In particular, the extract from P. eryngii induced high levels of IFN-γ and reduced levels of IL-4. We further investigated the use of a trial immunogen using the P. eryngii extract as a Th1 immunostimulator. An oil-in-water emulsion of the hot-water extract from P. eryngii (immunostimulator) and ovalbumin (OVA; antigen) was used as a trial immunogen. This immunogen induced strong OVA-specific IgG2a antibody production in mice compared with the negative controls. In addition, OVA-specific IgG1 antibody levels were lower than those for the negative controls. Marked increases in serum IFN-γ levels and high-level production of IFN-γ in the culture supernatant from the CD4(+) spleen cells in the trial immunogen group mice were observed. Our results suggested that the hot-water extract from P. eryngii induced Th1 immunity by acting as an immunostimulator.

Heteroglycan from an alkaline extract of a somatic hybrid mushroom (PfloVv1aFB) of Pleurotus florida and Volvariella volvacea: structural characterization and study of immunoenhancing properties.

A water soluble heteroglycan, isolated from the alkaline extract of the fruit bodies of the somatic hybrid mushroom (PfloVv1aFB), raised through protoplast fusion between the strains of Pleorutus florida and Volverilla volvacea, was found to consist of (1→3)-, (1→6)-, (1→3,4)-linked, and terminal ß-D-Glcp along with (1→2,6)-α-D-Galp and terminal α-D-Manp in a relative proportion of approximately 1:1:1:1:1:1. This polysaccharide exhibited strong immunostimulating activity of macrophages as well as splenocytes and thymocytes. Structural investigation was carried out using sugar analysis, methylation analysis; periodate oxidation study, and NMR experiments ((1)H, (13)C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC). On the basis of the above mentioned experiments, the structure of the repeating unit of the polysaccharide was established as: [formula see text].

Response to oyster mushroom (Pleurotus ostreatus) extract by sensitized and nonsensitized guinea pig trachea.

Occupational exposure to oyster mushroom (Pleurotus ostreatus) has been associated with obstructive lung disease. Previously, we studied an extract of oyster mushroom (OME) and determined that it causes dose-dependent contractions of nonsensitized guinea pig trachea (GPT). We extend these studies to the investigation of sensitized tissue. In the present study 24 animals were sensitized using ovalbumin (OA) and subsequently challenged with an aerosol of 2.5% OA. A control group of 12 nonsensitized GPs was also studied. Tracheas were removed and were divided into rings in which the epithelium was retained (EP+) or removed (EP-). Dose-related contractions of sensitized and nonsensitized GPTs were elicited with OME. In nonsensitized animals the EP+ GPTs demonstrated a significantly greater response to OME (100-1000 µl) than did the EP- GPTs (p < 0.01). By contrast, in sensitized GPTs with and without epithelium there was no difference to challenge with OME. Finally, sensitized GPTs with and without epithelium and nonsensitized GPTs with epithelium responded similarly to challenge with OME. These findings suggest that in nonsensitized animals there is an enhancement of contractile response to OME which is in part mediated by the GPT epithelium. In sensitized animals with or without epithelium, the level of bronchoconstrictor response is similar to that of the nonsensitized animals with epithelium, suggesting an enhanced constrictor response independent of epithelium in the sensitized animals.

Influencia del sustrato utilizado para el crecimiento de hongos comestibles sobre sus características nutraceúticas / The influence of the substrate used for growing edible fungi on their nutraceutic characteristics

Siendo Colombia un país en el cual hay un gran desarrollo de la actividad agraria, generadora de cantidades apreciables de desechos que dan lugar a un problema ambiental, el cultivo de los hongos presenta un potencial en aplicaciones biotecnológicas que derivan de la diversidad metabólica de los mismos. Dicho potencial está tanto en la producción de los hongos como alimentos con óptimas calidades nutricionales, como en la obtención a partir de las setas de productos bioactivos que permitirían en un futuro no muy lejano contribuir a la economía del país y a la descontaminación ambiental. Dentro del reino fungi, las especies de Pleurotus pueden crecer sobre una amplia variedad de sustratos y tienen especiales propiedades para degradar los componentes lignocelulósicos presentes en mayor proporción en los residuos de la agroindustria. El objetivo del presente estudio fue el de evaluar el efecto del sustrato sobre las propiedades nutricionales o nutriceúticas de hongos del género Pleurotus. Como resultado se determinó que efectivamente la composición de los hongos en cuanto al contenido de proteínas netas, fibra, humedad, cenizas, carbohidratos y grasas totales varía con el sustrato empleado, lo que incide directamente en las propiedades antes anotadas, si se tiene en cuenta que dentro de los componentes determinados se encuentran metabolitos como polisacáridos y esteroles con bioacciones previamente reportadas. De igual manera, se encontró que estas variaciones son diferentes dependiendo de la especie.

Being Colombia a country in which there is a great development of the generating agrarian activity of appreciable quantities of waste that they give place to an environmental problem, the cultivation of the mushrooms presents a potential in biotechnical applications that derive of the metabolic diversity of the same ones. This potential is in the production of the mushrooms like in foods with good nutritional qualities. For instance in the obtaining starting from the mushrooms of bioactive products that would not allow in a very distant future to contribute to the economy of the country and the environmental decontamination. Inside the fungi Kingdom, the Pleurotus species can grow about a wide substrate variety and they have special properties to degrade the lignocelulosic component present in bigger proportions in the residuals of the agroindustry. The objective of the present study was of evaluating the substrate effect on the nutritional and/or nutriceutical quality of mushrooms of the Pleurotus gender. As a result it was determined that indeed the composition of the mushrooms as for the content of net proteins, fiber, humidity, ashes, carbohydrates and total fats, varies with the substrate used, what impacts directly in their properties, taking into account that in certain components are metabolites such as polysaccharides and sterols with bioactions previously reported. Similarly it was found that these variations are different depending on the species.

[Hypersensitivity pneumonitis induced by Pleurotus eryngii spores--a case report].

A 52-year-old man was hospitalized because of recurrent fever, dyspnea and cough in February 2002. He had worked in a Eringi (Pleurotus eryngii) mushroom factory since 1996. Chest radiography showed diffuse fine nodular shadows. Chest computed tomography demonstrated centrilobular nodules and increased attenuation in both lungs. The patient suffered from hypoxemia (PaO2 = 65 torr) while breathing room air. The lymphocyte count in the bronchoalveolar lavage fluid was increased, and transbronchial lung biopsy specimens showed lymphocyte alveolitis with epithelioid cell granulomas in the alveolar spaces. After admission, the patient's symptoms improved rapidly without medication. However, on his return to work, fever and hypoxemia appeared again. The lymphocyte stimulating test was positive against extracts of Eringi spores. Precipitins against the extracts of Eringi spores were detected by the double immunodiffusion test. Our diagnosis was hypersensitivity pneumonitis (HP) caused by Eringi spores. In Japan, more than 30 cases of HP induced by mushroom spores have been reported. It is a matter of great urgency to prevent the occurrence of occupational HP in mushroom factories.

Hypersensitivity pneumonitis induced by the spore of Pleurotus Eryngii (Eringi).

We reported the first case of hypersensitivity pneumonitis (HP) by an edible mushroom, Pleurotus Eryngii (Eringi). A 54-year-old woman had worked in a Bunashimeji mushroom factory for 42 months, and she moved to a new factory producing Eringi. Two months after, she was found to have HP by the spore of Eringi. Although no radiological finding was detected 6 months before the onset of HP, serum surfactant protein D (SP-D) had been elevated. We speculated that type II pneumocyte activation might prepare the ground for HP during the former exposure to Bunashimeji, and serum SP-D levels might reflect their conditions.

Mushroom worker's lung resulting from indoor cultivation of Pleurotus osteatus.

Indoor cultivation of oyster mushroom Pleurotus osteatus lead to an outbreak of extrinsic allergic alveolitis in two workers. High titer of indirect fluorescent antibody and positive precipitins against basidiospores of P. osteatus were demonstrated in sera of the patients. Mushroom workers should protect themselves from the basidiospores, being aware of their pathogenicity.