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1.
Med Mycol ; 58(8): 1120-1125, 2020 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-32396166

RESUMEN

Pythium insidiosum is an oomycete that affects mammals, especially humans and horses, causing a difficult-to-treat disease. Typically, surgical interventions associated with antimicrobial therapy, immunotherapy, or both are the preferred treatment choices. PitiumVac® is a therapeutic vaccine prepared from the mycelial mass of P. insidiosum and is used to treat Brazilian equine pythiosis. To better understand how PitiumVac® works, we analyzed the composition of PitiumVac® and the immune response triggered by this immunotherapy in mice. We performed an enzymatic quantification that showed a total glucan content of 21.05% ± 0.94 (α-glucan, 6.37% ± 0.77 and (1,3)(1,6)-ß-glucan, 14.68% ± 0.60) and mannose content of 1.39% ± 0.26; the protein content was 0.52 mg ml-1 ± 0.07 mg ml-1. Healthy Swiss mice (n = 3) were subcutaneously preimmunized with one, two, or three shots of PitiumVac®, and immunization promoted a relevant Th1 and Th17 responses compared to nonimmunization of mice. The highest cytokine levels were observed after the third immunization, principally for IFN-γ, IL-17A, IL-6, and IL-10 levels. Results of infected untreated (Pythiosis) and infected treated (Pythiosis + PVAC) mice (n = 3) showed that PitiumVac® reinforces the Th1/Th17 response displayed by untreated mice. The (1,3)(1,6)-ß-glucan content can be, at least in part, related to this Th1/Th17 response.


Asunto(s)
Inmunoterapia , Pitiosis/terapia , Pythium/inmunología , Células TH1/inmunología , Células Th17/inmunología , Animales , Citocinas/inmunología , Glucanos/análisis , Glucanos/inmunología , Inmunización , Ratones , Micelio/química , Micelio/inmunología , Pitiosis/inmunología , Vacunas/administración & dosificación , Vacunas/química , Vacunas/inmunología
2.
FEMS Yeast Res ; 18(3)2018 05 01.
Artículo en Inglés | MEDLINE | ID: mdl-29462298

RESUMEN

Candida albicans is a human commensal that causes opportunistic infections. Th17 cells provide resistance against mucosal infection with C. albicans; however, the T cell antigens remain little known. Our final goal is to find effective T cell antigens of C. albicans that are responsible for immunotherapy against candidiasis. Here, we prepared fractions including cytosol, membrane and cell wall from yeast and mycelial cells. Proteins derived from a membrane fraction of mycelial cells effectively induced differentiation of CD4+ T cells into IL-17A-producing Th17 cells. To confirm the immunological response in vivo of proteins from mycelial membrane, we performed adoptive transfer experiments using ex vivo stimulated CD4+ T cells from IL-17A-GFP reporter mice. Mycelial membrane-differentiated CD4+ Th17 cells adoptively transferred intravenously prevented oral candidiasis by oral infection of C. albicans, compared with control anti-CD3-stimulated CD4+ T cells. This was confirmed by the clinical score and the number of neutrophils on the infected tissues. These data suggest that effective T cell antigens against candidiasis could be present in the membrane protein fraction of mycelial cells. The design of novel vaccination strategies against candidiasis will be our next step.


Asunto(s)
Candidiasis Bucal/prevención & control , Proteínas Fúngicas/farmacología , Micelio/química , Células Th17/inmunología , Traslado Adoptivo , Animales , Antígenos Fúngicos/inmunología , Antígenos Fúngicos/farmacología , Candida albicans/inmunología , Candidiasis Bucal/inmunología , Diferenciación Celular , Femenino , Proteínas Fúngicas/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Micelio/inmunología , Células Th17/citología
3.
J Mycol Med ; 28(2): 285-288, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29482975

RESUMEN

PURPOSE: The aim of this study was to develop and evaluate the diagnostic potential of indirect enzyme-linked immunosorbent assay (I-ELISA) for the rapid and precise diagnosis of Microsporum canis infection in humans. BASIC PROCEDURES: The present study reports the production, partial purification and SDS-PAGE analysis of M. canis mycelial antigens and production of specific polyclonal antibodies. It also reports the development and optimization of indirect ELISA and evaluation of its potential for the diagnosis of M. canis infection in humans. MAIN FINDINGS: An I-ELISA showed the sensitivity of 94.55% and specificity of 93.33%. Positive and negative predictive values were 96.30% and 90.32% respectively. Receiver operating characteristic (ROC) curve analysis of the data showed higher diagnostic accuracy. The area under the curve (AUC) was 0.925. A significant correlation coefficient of 0.8771 (P<0.0001) was obtained between I-ELISA and fungal culture method. PRINCIPAL CONCLUSIONS: In conclusion, the present study clearly shows the detection of specific antibodies by indirect ELISA using M. canis antigens. The assay is sensitive, specific and easy to perform, could enable rapid and more convenient diagnosis of dermatophytosis in humans.


Asunto(s)
Dermatomicosis/diagnóstico , Ensayo de Inmunoadsorción Enzimática/métodos , Microsporum/aislamiento & purificación , Pruebas Serológicas/métodos , Antígenos Fúngicos/inmunología , Dermatomicosis/sangre , Pruebas de Enzimas/métodos , Humanos , Micelio/inmunología , Curva ROC , Sensibilidad y Especificidad
4.
Artículo en Inglés | MEDLINE | ID: mdl-26264521

RESUMEN

The host pulmonary response to the fungus Histoplasma capsulatum was evaluated, through the profile of cytokines detected by the MagPix magnetic beads platform in lung homogenates and by lung-granulomas formation, from mice intra-nasally infected with mycelial propagules (M-phase) of two virulent H. capsulatum strains, EH-46 and G-217B. Results highlight that mice lung inflammatory response depends on the H. capsulatum strain used, during the first step of the fungal infection. IL-1ß and TNF-α increased their concentrations in mice infected with both strains. The highest levels of IL-6, IL-17, and IL-23 were found in EH-46-infected mice, whereas levels of IL-22 were variable at all post-infection times for both strains. Significant increases of IL-12, IFN-γ, IL-4, and IL-10 were associated to EH-46-infected mice. Histological lung findings from EH-46-infected mice revealed incipient and numerous well-developed granulomas, distributed in lung-lobes at the 14th and the 21st days after infection, according to cytokine profiles.


Asunto(s)
Citocinas/inmunología , Citocinas/metabolismo , Histoplasma/inmunología , Histoplasmosis/inmunología , Pulmón/inmunología , Animales , Histoplasma/crecimiento & desarrollo , Histoplasma/patogenicidad , Histoplasmosis/microbiología , Interferón gamma/inmunología , Interferón gamma/metabolismo , Interleucina-12/inmunología , Interleucina-12/metabolismo , Interleucina-6/inmunología , Interleucina-6/metabolismo , Interleucinas/inmunología , Interleucinas/metabolismo , Pulmón/microbiología , Pulmón/patología , Pulmón/ultraestructura , Masculino , Ratones , Ratones Endogámicos BALB C , Micelio/inmunología , Micelio/patogenicidad , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Interleucina-22
5.
Rev Iberoam Micol ; 31(1): 76-80, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24252831

RESUMEN

ß-1,3-Glucan is important for infective forms (mycelial phase) of Histoplasma capsulatum and shares many features allotted to pathogen-associated molecular patterns. These cell wall carbohydrates interact with phagocytes by binding to Toll and lectin-like receptors, present on cell surfaces of macrophages, neutrophils, and dendritic cells. This review focuses on recent findings of the major H. capsulatum and host carbohydrate-driven interactions that account for internalization of fungal infective forms into phagocytes, and its subsequent avoidance of intracellular elimination. The yeast phase of H. capsulatum possesses different modulating factors of the macrophagic-anti-fungal mechanisms, mainly α-1,3-glucan, which is considered relevant for virulence. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012).


Asunto(s)
Glucanos/inmunología , Histoplasma/inmunología , Histoplasmosis/inmunología , Interacciones Huésped-Patógeno/inmunología , Inmunidad Innata , beta-Glucanos/inmunología , Conformación de Carbohidratos , Secuencia de Carbohidratos , Pared Celular , Células Dendríticas/inmunología , Humanos , Lectinas/inmunología , Macrófagos/inmunología , Datos de Secuencia Molecular , Micelio/inmunología , Neutrófilos/inmunología , Fagocitos/fisiología , Fagocitosis , Receptores Mitogénicos/inmunología , Receptores Toll-Like/inmunología , Virulencia/inmunología
6.
J Agric Food Chem ; 61(22): 5207-14, 2013 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-23638999

RESUMEN

Preparations with elicitation activity were obtained from the mycelium of Leptosphaeria maculans , a fungal pathogen of oilseed rape (Brassica napus). Crude delipidated and deproteinized extract from fungal cell walls induced expression of pathogenesis related gene 1 (PR1), hydrogen peroxide accumulation, and enhanced resistance of B. napus plants toward infection by L. maculans. Elicitation activity significantly decreased after treatment of a crude extract with α- or ß-glucanase. Monosaccharide composition analysis of a crude extract purified by ion-exchange chromatography revealed glucose (∼58 mol %), mannose (∼22 mol %), and galactose (∼18 mol %) as the major sugars. FT-IR and NMR spectra confirmed the presence of both carbohydrate and polypeptide components in the purified product. Correlation NMR experiments defined trisaccharide bound to O-3 of serine residue α-D-Glcp-(1→2)-ß-D-Galf-(1→6)-α-D-Manp-(1→3)-L-Ser. Terminal α-D-Glcp and (1→6)-ß-D-glucan were also detected. The obtained results strongly support the conclusion that these carbohydrates induce defense response in B. napus plants.


Asunto(s)
Ascomicetos/química , Brassica napus/efectos de los fármacos , Extractos Celulares/farmacología , Pared Celular/química , Resistencia a la Enfermedad/efectos de los fármacos , Fungicidas Industriales/farmacología , Regulación hacia Arriba/efectos de los fármacos , Ascomicetos/crecimiento & desarrollo , Ascomicetos/inmunología , Aspergillus niger/enzimología , Brassica napus/inmunología , Brassica napus/metabolismo , Brassica napus/microbiología , Extractos Celulares/química , Extractos Celulares/aislamiento & purificación , Química Agrícola/métodos , Regulación hacia Abajo , Proteínas Fúngicas/análisis , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Fungicidas Industriales/química , Fungicidas Industriales/aislamiento & purificación , Glicósido Hidrolasas/biosíntesis , Glicósido Hidrolasas/genética , Glicósido Hidrolasas/metabolismo , Glicósidos/análisis , Glicósidos/química , Glicósidos/metabolismo , Peróxido de Hidrógeno/metabolismo , Hidrólisis , Micelio/química , Micelio/crecimiento & desarrollo , Micelio/inmunología , Oligosacáridos/análisis , Oligosacáridos/química , Oligosacáridos/metabolismo , Fragmentos de Péptidos/análisis , Fragmentos de Péptidos/química , Fragmentos de Péptidos/metabolismo , Proteínas de Plantas/biosíntesis , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantones/efectos de los fármacos , Plantones/inmunología , Plantones/metabolismo , Plantones/microbiología
7.
J Sci Food Agric ; 93(12): 3081-6, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-23526300

RESUMEN

BACKGROUND: Soybean sprouts (Kongnamool) are one of the most popular and nutritive traditional vegetables in East Asia. Anthracnose caused by Colletotrichum gloeosporioides is one of the most serious diseases of soybean sprouts. In order to obtain basic information for breeding and/or selecting soybean genotypes with increased natural defense against anthracnose, phenolic compounds were profiled for healthy and infected soybean (Glycine max Merr.) sprouts by using high-performance liquid chromatography coupled with tandem mass spectrometry. RESULTS: Tryptophan and eight phenolic compounds (daidzin, genistin, malonyldaidzin, malonylgenistin, daidzein, glycitein, genistein and coumestrol) were determined from healthy and inoculated sprouts. Total identified phenolic content was 40.02 ± 0.03 mg kg⁻¹, 99.4% of which was isoflavones. CONCLUSION: The monitoring suggested that de novo induced glycitein appeared to act as a phytoalexin in the defence mechanism of the soybean sprouts against C. gloeosporioides, and constitutively formed seven phenolic components that functioned as phytoanticipins in the diseased soybean sprouts.


Asunto(s)
Colletotrichum/crecimiento & desarrollo , Glycine max/metabolismo , Glycine max/microbiología , Isoflavonas/biosíntesis , Enfermedades de las Plantas/microbiología , Plantones/metabolismo , Plantones/microbiología , Antifúngicos/análisis , Antifúngicos/química , Antifúngicos/metabolismo , Antifúngicos/farmacología , Cromatografía Líquida de Alta Presión , Colletotrichum/efectos de los fármacos , Colletotrichum/inmunología , Germinación , Glucósidos/análisis , Glucósidos/biosíntesis , Glucósidos/química , Glucósidos/farmacología , Isoflavonas/análisis , Isoflavonas/química , Isoflavonas/farmacología , Pruebas de Sensibilidad Microbiana , Micelio/efectos de los fármacos , Micelio/crecimiento & desarrollo , Micelio/inmunología , Fenoles/análisis , Fenoles/química , Fenoles/metabolismo , Fenoles/farmacología , Enfermedades de las Plantas/inmunología , Inmunidad de la Planta , República de Corea , Plantones/crecimiento & desarrollo , Plantones/inmunología , Sesquiterpenos/análisis , Sesquiterpenos/química , Sesquiterpenos/metabolismo , Sesquiterpenos/farmacología , Glycine max/crecimiento & desarrollo , Glycine max/inmunología , Espectrometría de Masa por Ionización de Electrospray , Esporas Fúngicas/efectos de los fármacos , Esporas Fúngicas/crecimiento & desarrollo , Esporas Fúngicas/inmunología , Espectrometría de Masas en Tándem , Fitoalexinas
8.
Plant Cell Rep ; 31(12): 2121-37, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22833277

RESUMEN

UNLABELLED: Wheat cultivar Xingzi 9104 (XZ) possesses adult plant resistance (APR) to stripe rust caused by Puccinia striiformis f. sp. tritici (Pst). In this study, histological and cytological experiments were conducted to elucidate the mechanisms of APR in XZ. The results of leaf inoculation experiments indicated that APR was initiated at the tillering stage, gradually increased as the plant aged and highly expressed after boot stage. The histology and oxidative burst in infected leaves of plants at seedling, tillering and boot stages were examined using light microscopic and histochemical methods. Subcellular changes in the host-pathogen interactions during the seedling and boot stages were analyzed by transmission electron microscopy. The results showed that haustorium formation was retarded in the adult plants and that the differentiation of secondary intercellular hyphae was significantly inhibited, which decreased the development of microcolonies in the adult plants, especially in plants of boot stage. The expression of APR to stipe rust during wheat development was clearly associated with extensive hypersensitive cell death of host cells and localized production of reactive oxygen species, which coincided with the restriction of fungal growth in infection sites in adult plants. At the same time, cell wall-related resistance in adult plants prevented ingression of haustorial mother cells into plant cells. Haustorium encasement was coincident with malformation or death of haustoria. The results provide useful information for further determination of mechanisms of wheat APR to stripe rust. KEY MESSAGE: The expression of APR to stipe rust in wheat cultivar Xingzi 9104 (XZ) was clearly associated with extensive hypersensitive cell death of host cells and the localized production of reactive oxygen species.


Asunto(s)
Basidiomycota/patogenicidad , Resistencia a la Enfermedad , Interacciones Huésped-Patógeno , Hojas de la Planta/microbiología , Triticum/citología , Muerte Celular , Histocitoquímica , Peróxido de Hidrógeno/metabolismo , Microscopía Electrónica de Transmisión , Micelio/inmunología , Micelio/ultraestructura , Oxidación-Reducción , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Hojas de la Planta/inmunología , Hojas de la Planta/ultraestructura , Especies Reactivas de Oxígeno/metabolismo , Plantones/inmunología , Plantones/microbiología , Factores de Tiempo , Triticum/inmunología , Triticum/metabolismo , Triticum/microbiología
9.
J Proteome Res ; 11(5): 2739-53, 2012 May 04.
Artículo en Inglés | MEDLINE | ID: mdl-22424419

RESUMEN

An immune response is triggered in host cells when host receptors recognize conserved molecular motifs, pathogen-associated molecular patterns (PAMPs), such as ß-glucans, and chitin at the cell surface of a pathogen. Effector-triggered immunity occurs when pathogens deliver effectors into the host cell to suppress the first immune signaling. Using a differential proteomic approach, we identified an array of proteins responding to aflatoxins in cotyledons of peanut (Arachis hypogaea) infected with aflatoxin-producing (toxigenic) but not nonaflatoxin-producing (atoxigenic) strains of Aspergillus flavus. These proteins are involved in immune signaling and PAMP perception, DNA and RNA stabilization, induction of defense, innate immunity, hypersensitive response, biosynthesis of phytoalexins, cell wall responses, peptidoglycan assembly, penetration resistance, condensed tannin synthesis, detoxification, and metabolic regulation. Gene expression analysis confirmed the differential abundance of proteins in peanut cotyledons supplemented with aflatoxins, with or without infection with the atoxigenic strain. Similarly, peanut germination and A. flavus growth were altered in response to aflatoxin B1. These findings show an additional immunity initiated by aflatoxins. With the PAMP- and effector-triggered immune responses, this immunity constitutes the third immune response of the immune system in peanut cotyledon cells. The system is also a three-grade coevolution of plant-pathogen interaction.


Asunto(s)
Aflatoxina B1/inmunología , Arachis/inmunología , Aspergillus flavus/patogenicidad , Inmunidad de la Planta , Proteoma/análisis , Aflatoxina B1/genética , Arachis/genética , Arachis/microbiología , Pared Celular , Clonación Molecular , Cotiledón/genética , Cotiledón/inmunología , Cotiledón/microbiología , Regulación de la Expresión Génica de las Plantas , Germinación , Interacciones Huésped-Patógeno , Micelio/crecimiento & desarrollo , Micelio/inmunología , Células Vegetales/inmunología , Células Vegetales/microbiología , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Proteínas de Almacenamiento de Semillas/inmunología , Semillas/inmunología , Semillas/microbiología , Transducción de Señal , Especificidad de la Especie
10.
J Proteome Res ; 11(1): 172-88, 2012 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-22142336

RESUMEN

Pseudallescheria boydii is a filamentous fungus that causes a wide array of infections that can affect practically all the organs of the human body. The treatment of pseudallescheriosis is difficult since P. boydii exhibits intrinsic resistance to the majority of antifungal drugs used in the clinic and the virulence attributes expressed by this fungus are unknown. The study of the secretion of molecules is an important approach for understanding the pathogenicity of fungi. With this task in mind, we have shown that mycelial cells of P. boydii were able to actively secrete proteins into the extracellular environment; some of them were recognized by antibodies present in the serum of a patient with pseudallescheriosis. Additionally, molecules secreted by P. boydii induced in vitro irreversible damage in pulmonary epithelial cells. Subsequently, two-dimensional gel electrophoresis combined with mass spectrometry was carried out in order to start the construction of a map of secreted proteins from P. boydii mycelial cells. The two-dimensional map showed that most of the proteins (around 100 spots) were focused at pH ranging from 4 to 7 with molecular masses ranging from 14 to >117 kDa. Fifty spots were randomly selected, of which 30 (60%) were consistently identified, while 20 (40%) spots generated peptides that showed no resemblance to any known protein from other fungi and/or MS with low quality. Notably, we identified proteins involved in metabolic pathways (energy/carbohydrate, nucleotide, and fatty acid), cell wall remodeling, RNA processing, signaling, protein degradation/nutrition, translation machinery, drug elimination and/or detoxification, protection against environmental stress, cytoskeleton/movement proteins, and immunogenic molecules. Since the genome of this fungus is not sequenced, we performed enzymatic and immunodetection assays in order to corroborate the presence of some released proteins. The identification of proteins actively secreted by P. boydii provides important new information for understanding immune modulation and provides important new perspectives on the biology of this intriguing fungus.


Asunto(s)
Proteínas Fúngicas/metabolismo , Genoma Fúngico , Micelio/metabolismo , Micosis/microbiología , Proteoma/metabolismo , Pseudallescheria/metabolismo , Secuencia de Aminoácidos , Anticuerpos Antifúngicos/sangre , Antígenos Fúngicos/inmunología , Línea Celular Tumoral , Electroforesis en Gel Bidimensional , Proteínas Fúngicas/química , Proteínas Fúngicas/inmunología , Proteínas Fúngicas/farmacología , Humanos , Concentración 50 Inhibidora , Viabilidad Microbiana , Datos de Secuencia Molecular , Micelio/crecimiento & desarrollo , Micelio/inmunología , Micelio/ultraestructura , Micosis/sangre , Fragmentos de Péptidos/química , Mapeo Peptídico , Proteoma/química , Proteoma/inmunología , Proteoma/farmacología , Proteómica , Pseudallescheria/crecimiento & desarrollo , Pseudallescheria/inmunología , Pseudallescheria/ultraestructura
11.
Int J Med Mushrooms ; 13(3): 237-44, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-22135875

RESUMEN

Treatment of hot water extract of the sclerotium of Polyporus rhinocerus (PRW) with murine macrophages including RAW 264.7 cell line and primary macrophages (PMs) could enhance their functional activities. These include a significant up-regulation of pinocytosis; an increase in the production of reactive oxygen species (ROS) and nitric oxide (NO); an increase in tumor necrosis factor alpha (TNF-alpha) production and inducible nitric oxide synthase (iNOS) expression in both RAW 264.7 cells and PMs. Cell surface receptors for yeast-derived beta-glucan, including Dectin-1, CR3, and TLR2, were determined by flow cytometry, and the expression of Dectin-1+ cells on the cell surface decreased in the responses of PMs to PRW. PRW increased phosphorylation of IkappaBalpha, which could trigger the nuclear factor kappa B (NF-kappaB) signal pathway for macrophage activation in RAW 264.7 cells. Therefore, the immunomodulatory effect of PRW could be mediated by macrophage activation via the NF-kappaB signal pathway.


Asunto(s)
Mezclas Complejas/farmacología , Activación de Macrófagos/efectos de los fármacos , Macrófagos/inmunología , FN-kappa B/metabolismo , Polyporus/química , Animales , Línea Celular , Mezclas Complejas/química , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Proteínas I-kappa B/metabolismo , Proteínas I-kappa B/farmacología , Lectinas Tipo C/metabolismo , Antígeno de Macrófago-1/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Micelio/química , Micelio/inmunología , Inhibidor NF-kappaB alfa , FN-kappa B/antagonistas & inhibidores , FN-kappa B/genética , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosforilación , Pinocitosis/efectos de los fármacos , Polyporus/inmunología , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 2/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia Arriba/efectos de los fármacos , beta-Glucanos/metabolismo
12.
Mycoses ; 54(5): e474-80, 2011 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-21554418

RESUMEN

The action of the complement system on pigmented and hypopigmented mycelia of the fungus Fonsecaea pedrosoi, the major aetiological pathogen of the chromoblastomycosis is herein discussed. Fungi were grown in medium Czapeck-Dox at 37°C, for 14 days, without shaking to obtain pigmented mycelium. To obtain hypopigmented mycelium, the fungus was grown at the same conditions, but in the dark and with low oxygenation. Activation was measured by complement consumption and enzyme-linked immunosorbent assay. We also observed by immunofluorescence the deposition of C3, C4 fragments and C9 on the surface of the different forms studied. The results indicate that both forms were able to activate the complement system mainly by the alternative pathway. Pigmented mycelia had the highest consumption results, indicating that the pigment, melanin, may have influence in activation.


Asunto(s)
Ascomicetos/inmunología , Activación de Complemento , Proteínas del Sistema Complemento/inmunología , Micelio/inmunología , Adulto , Medios de Cultivo/química , Oscuridad , Ensayo de Inmunoadsorción Enzimática , Experimentación Humana , Humanos , Microscopía Fluorescente , Micología/métodos , Pigmentos Biológicos/metabolismo
13.
J Med Virol ; 82(1): 128-37, 2010 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-19950232

RESUMEN

The identification of a safe and effective adjuvant that is able to enhance mucosal immune responses is necessary for the development of an efficient inactivated intranasal influenza vaccine. The present study demonstrated the effectiveness of extracts of mycelia derived from edible mushrooms as adjuvants for intranasal influenza vaccine. The adjuvant effect of extracts of mycelia was examined by intranasal co-administration of the extracts and inactivated A/PR8 (H1N1) influenza virus hemagglutinin (HA) vaccine in BALB/c mice. The inactivated vaccine in combination with mycelial extracts induced a high anti-A/PR8 HA-specific IgA and IgG response in nasal washings and serum, respectively. Virus-specific cytotoxic T-lymphocyte responses were also induced by administration of the vaccine with extract of mycelia, resulting in protection against lethal lung infection with influenza virus A/PR8. In addition, intranasal administration of NIBRG14 vaccine derived from the influenza A/Vietnam/1194/2004 (H5N1) virus strain administered in conjunction with mycelial extracts from Phellinus linteus conferred cross-protection against heterologous influenza A/Indonesia/6/2005 virus challenge in the nasal infection model. In addition, mycelial extracts induced proinflammatory cytokines and CD40 expression in bone marrow-derived dendritic cells. These results suggest that mycelial extract-adjuvanted vaccines can confer cross-protection against variant H5N1 influenza viruses. The use of extracts of mycelia derived from edible mushrooms is proposed as a new safe and effective mucosal adjuvant for use for nasal vaccination against influenza virus infection.


Asunto(s)
Adyuvantes Inmunológicos , Agaricales , Glicoproteínas Hemaglutininas del Virus de la Influenza , Subtipo H5N1 del Virus de la Influenza A/patogenicidad , Vacunas contra la Influenza , Micelio , Infecciones por Orthomyxoviridae , Vacunas de Productos Inactivados , Adyuvantes Inmunológicos/administración & dosificación , Administración Intranasal , Agaricales/crecimiento & desarrollo , Agaricales/inmunología , Animales , Anticuerpos Antivirales/sangre , Reacciones Cruzadas/efectos de los fármacos , Reacciones Cruzadas/inmunología , Glicoproteínas Hemaglutininas del Virus de la Influenza/administración & dosificación , Glicoproteínas Hemaglutininas del Virus de la Influenza/inmunología , Humanos , Inmunidad/efectos de los fármacos , Inmunidad/inmunología , Inmunidad Mucosa , Inmunoglobulina A Secretora/metabolismo , Inmunoglobulina G/sangre , Subtipo H1N1 del Virus de la Influenza A/inmunología , Subtipo H1N1 del Virus de la Influenza A/metabolismo , Subtipo H5N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/administración & dosificación , Vacunas contra la Influenza/inmunología , Ratones , Ratones Endogámicos BALB C , Micelio/inmunología , Infecciones por Orthomyxoviridae/inmunología , Infecciones por Orthomyxoviridae/mortalidad , Infecciones por Orthomyxoviridae/prevención & control , Infecciones por Orthomyxoviridae/virología , Linfocitos T/inmunología , Vacunas de Productos Inactivados/administración & dosificación , Vacunas de Productos Inactivados/inmunología
14.
J Immunotoxicol ; 6(1): 62-73, 2009 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-19519164

RESUMEN

Intratracheal aspiration (IA) exposure to Metarhizium anisopliae crude antigen (MACA), which is composed of equal protein amounts of mycelium (MYC), conidia (CON) and inducible proteases/chitinases (IND) extracts/filtrates, has resulted in responses characteristic of human allergic asthma in mice. The study objective was to evaluate the potential of each component extract to induce allergic/asthma-like responses observed in this mouse model. BALB/c mice received 4 IA exposures to MACA, CON, MYC, IND, or bovine serum albumin (BSA; negative control) or appropriate vehicle control or inflammatory control over a 4-wk period. Mice were assessed by whole-body plethysmography for immediate airway responses and airway hyperresponsiveness to methacholine (Mch) challenge (PenH). Serum and bronchoalveolar lavage fluid (BALF) were collected 3 d after the final exposure. Additionally, BALF neurotrophin levels and extract protease and chitinase activity levels were evaluated. Western blot analysis showed that each component contained different IgE-reactive proteins. All fungal extract exposures resulted in elevated BALF total and differential cell counts, IgE and IgA and total serum IgE compared to HBSS and BSA controls. MYC-exposed mice had the highest responses except for neutrophil influx, which was highest in MACA and IND exposures. However, the MYC-exposed mice had significantly lower PenH values compared to other treatments. By comparison IND and MACA induced significantly higher PenH values. Additionally, IND had substantially higher protease activity levels but induced the lowest neurotrophin levels compared to the other fungal exposures. In this allergic asthma model extract chitinase activity was not associated with allergic responses. In summary, multiple exposures to any of the M. anisopliae component extracts induced allergic/asthma-like responses in BALB/c mice but the response magnitude was different for each component and each appears to contain unique IgE-reactive proteins. Therefore, hazard identification and/or risk assessment for molds must test both mycelia and conidia.


Asunto(s)
Antígenos/inmunología , Asma/inmunología , Metarhizium/inmunología , Animales , Antígenos/administración & dosificación , Antígenos/química , Asma/inducido químicamente , Asma/patología , Asma/fisiopatología , Hiperreactividad Bronquial/inmunología , Hiperreactividad Bronquial/fisiopatología , Pruebas de Provocación Bronquial , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Recuento de Células , Enzimas/administración & dosificación , Enzimas/química , Enzimas/inmunología , Femenino , Granulocitos/citología , Granulocitos/inmunología , Inmunoglobulina A/inmunología , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Pulmón/fisiopatología , Linfocitos/citología , Linfocitos/inmunología , Metarhizium/química , Metarhizium/enzimología , Ratones , Ratones Endogámicos BALB C , Micelio/química , Micelio/inmunología , Factores de Crecimiento Nervioso/análisis , Factores de Crecimiento Nervioso/metabolismo , Plaguicidas/química , Plaguicidas/inmunología , Esporas Fúngicas/química , Esporas Fúngicas/inmunología , Vacunación
15.
Clin Exp Med ; 9(4): 277-84, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19350364

RESUMEN

Mycelia products from wild-form Cordyceps sinensis could be constantly produced in a large scale and would be a better source of this herbal medicine. Our purpose was to investigate the immunological effects of an orally administered hot-water extract cultured mycelium of C. sinensis in lupus-prone (NZB/NZW) F1 hybrids. Forty female mice were divided into four groups and were given 2.4 mg/g/day oral doses of C. sinensis starting at three (group A), six (group B), or eight (group C) months of age, whereas the remaining group (group D) served as a control. Survival, proteinuria, and titers of anti-double-stranded DNA autoantibodies were evaluated. Treatment with C. sinensis resulted in increased survival, decreased proteinuria, and reduced titers of anti-double-stranded DNA antibody in groups A and B. Moreover, the mice in groups A and B showed significantly reduced percentages of CD4(+) T cells (*P < 0.05) and increased percentages of CD8(+) T cells in peripheral blood mononuclear cells (PBMC) after C. sinensis administration. At 6 months of age, the proliferation rate of BrdU-incorporated spleen cells was significantly decreased after 48 and 72 h of C. sinensis treatment (**P < 0.01) in group A of mice. In conclusions, early medication with C. sinensis induced the redistribution of PBMC and attenuated the disease severity of lupus in (NZB/NZW) F1 mice.


Asunto(s)
Cordyceps/inmunología , Medicamentos Herbarios Chinos/uso terapéutico , Lupus Eritematoso Sistémico/inmunología , Lupus Eritematoso Sistémico/prevención & control , Micelio/inmunología , Linfocitos T/inmunología , Administración Oral , Animales , Anticuerpos Antinucleares/inmunología , Autoanticuerpos/inmunología , Cartilla de ADN/química , Femenino , Lupus Eritematoso Sistémico/patología , Ratones , Ratones Endogámicos NZB , Proteinuria , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/patología , Tasa de Supervivencia
16.
Mycopathologia ; 165(4-5): 313-30, 2008.
Artículo en Inglés | MEDLINE | ID: mdl-18777636

RESUMEN

Knowledge concerning the host-Paracoccidioides brasiliensis interactions is abundant. Yet, most of the experimental studies have used yeast cells to prepare the corresponding inoculum. As these cells do not represent the naturally infecting propagules, the corresponding experiments by-pass the earlier stages of such interactions. Studies done in patients, who also harbour yeast cells, suffer from the same bias. The review presented below focuses on the immune responses of BALB/c mice infected with conidia obtained from P. brasiliensis mycelial form cultures, the fungal stage most probably existing in nature. As such, the corresponding experiments would copy the onset and course of the human infection. A large number of experimental studies done by the CIB Medical and Experimental Mycology Unit in a period of almost 25 years have been revised and extracted so as to present a comprehensive record on the immune responses induced when mice are infected intranasally with the conidia. The establishment of this mouse model has permitted the analysis of the immune responses taking place during the early and late stages post-challenge. This unique model has made possible to characterize the course of the experimental disease including the inflammatory reaction, the expression of cytokines and of the various molecules associated to these responses, all of which lead to granuloma formation. The latter structure serves as a nest for the development of fibrosis. Thus, we have also obtained a glimpse on the complexity that accompanies the fibrosis, the most common sequelae of paracoccidioidomycosis. Additionally, a concerted effort has been made to appraise the whole gamut of immune factors and related molecules that directly or indirectly, contribute to shape the pathogenesis of this Latin American mycosis.


Asunto(s)
Modelos Animales de Enfermedad , Enfermedades Pulmonares Fúngicas/inmunología , Pulmón/inmunología , Micelio/inmunología , Paracoccidioides/inmunología , Paracoccidioidomicosis/inmunología , Animales , Humanos , Pulmón/microbiología , Enfermedades Pulmonares Fúngicas/microbiología , Masculino , Ratones , Ratones Endogámicos BALB C , Micelio/patogenicidad , Paracoccidioides/crecimiento & desarrollo , Paracoccidioides/patogenicidad , Paracoccidioidomicosis/microbiología
17.
Int Immunopharmacol ; 8(6): 909-15, 2008 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-18442797

RESUMEN

Two proteoglycans, PNW1 and PNM1, were isolated from the mycelium of Phellinus nigricans through submerged fermentation and culture medium, respectively. PNW1 and PNM1 with similar average molecular weight (33 kDa and 29 kDa) were composed of glucose, galactose, mannose, arabinose and fucose in the molar ratios of 3.26:8.77:6.44:1:1.35 and 20.06:8.72:6.94:1:0.76. At the dose of 100, 200, and 400 mg/kg, PNW1 and PNM1 exhibited anti-tumor activity against mice-transplanted Sarcoma 180 in vivo. However, no direct cytotoxic activity against Sarcoma 180 could be determined. Significant increase in the relative spleen and thymus weight and expression of tumor necrosis factor-alpha (TNF-alpha) in serum was observed, decreasing the tumor weight significantly. PNW1 and PNM1 could stimulate lymphocytes proliferation and increase production of nitric oxide (NO) and TNF-alpha in macrophages. The results indicate that both lymphocyte and macrophages were activated by preparations of proteoglycans from mycelium and culture medium of P. nigricans. The anti-tumor effect of the proteoglycans is not directly tumoricidal but rather immunostimulating.


Asunto(s)
Basidiomycota/inmunología , Factores Inmunológicos , Linfocitos/inmunología , Macrófagos/inmunología , Proteoglicanos/inmunología , Sarcoma 180/inmunología , Animales , Factores Inmunológicos/farmacología , Activación de Linfocitos , Linfocitos/efectos de los fármacos , Linfocitos/metabolismo , Activación de Macrófagos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Micelio/inmunología , Óxido Nítrico/metabolismo , Fagocitosis , Proteoglicanos/farmacología , Sarcoma 180/metabolismo , Factor de Necrosis Tumoral alfa/sangre
18.
Wei Sheng Wu Xue Bao ; 47(4): 628-33, 2007 Aug.
Artículo en Chino | MEDLINE | ID: mdl-17944362

RESUMEN

Five Ganoderma species, including G. lucidum, G. tsugae, G. oerstedii, G. resinaceum and G. subamboinens, were parallel studied under an identical condition. These species were cultivated using liquid fermentation and their mycelia polysaccharides were extracted and compared on the physical/chemical properties and in vitro immunomodulatory activities. These results showed that the polysaccharide yields varied markedly among different species, and G. oerstedii was the highest among the five. However, HPLC analysis showed all the polysaccharide extracts had similar molecular weight distributions and monosaccharide compositions. They all contained glucose, galactose, mannose, glucosamine hydrochloride and fucose. In vitro assays, these polysaccharide extracts significantly stimulated phagocytosis and nitric oxide production by RAW 264.7 macrophage cell line, and G. subamboinens exerted the strongest potency. When Con A was not or presented, they all showed an up-or-down immunomodulatory effect on mouse splenocyte proliferation. The results illustrate that in addition to G. lucidum and G. tsugae, which are the two mostly studied and applied species, other Ganoderma species can also produce polysaccharides with similar physical/chemical properties and with similar immunomodulatory activities.


Asunto(s)
Ganoderma/química , Ganoderma/inmunología , Polisacáridos/química , Polisacáridos/inmunología , Animales , Línea Celular , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos BALB C , Peso Molecular , Micelio/química , Micelio/inmunología , Polisacáridos/farmacología
19.
Food Chem Toxicol ; 45(2): 278-85, 2007 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-17029726

RESUMEN

Streptococcal pyrogenic exotoxin B (SPE B) is a virulent factor in group A streptococcal infection. We previously showed that SPE B reduced phagocytosis in human monocytic U937 cells. Here we show that the mycelium extract of Cordyceps sinensis (CS), a Chinese immunomodulatory herbal medicine, increased phagocytosis in U937 cells. Neither heat nor trypsin pretreatment prevented CS extract from causing this increase. Further studies indicated that SPE B-mediated suppression of U937 cell phagocytic activity was abrogated by CS extract. Factors in the conditioned medium from CS-extract-treated U937 cells were responsible for blocking the SPE B-mediated suppression of phagocytosis. Heating the conditioned medium eliminated the increase, which suggested that the U937-cell protein products augmented phagocytosis. Analyzing cytokine mRNA expression of U937 cells revealed increases in interferon-gamma (IFN-gamma), interleukin (IL)-12 p35 and p40, and tumor necrosis factor-alpha (TNF-alpha), but not in IL-1beta, IL-6, or IL-8. Treating U937 cells with anti-IFN-gamma, IL-12, and TNF-alpha antibodies also eliminated the conditioned medium-induced increase in phagocytosis. Taken together, SPE B inhibited phagocytosis, but CS mycelium extract abrogated this inhibition by causing cytokine production.


Asunto(s)
Proteínas Bacterianas/toxicidad , Cordyceps/inmunología , Citocinas/biosíntesis , Exotoxinas/toxicidad , Inmunosupresores/farmacología , Fagocitosis/efectos de los fármacos , Proteínas Bacterianas/antagonistas & inhibidores , Cordyceps/química , Cordyceps/metabolismo , Citocinas/inmunología , Citocinas/farmacología , Exotoxinas/antagonistas & inhibidores , Fluorescencia , Calor , Humanos , Micelio/química , Micelio/inmunología , Micelio/metabolismo , Streptococcus pyogenes/metabolismo , Tripsina/farmacología , Células U937
20.
Allergol Immunopathol (Madr) ; 34(2): 64-9, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-16606548

RESUMEN

BACKGROUND: The lack of well standardized or characterized extracts that contain the relevant allergens of the appropriate fungus is resulting in a high heterogeneity of the commercial preparation. MATERIAL AND METHODS: Immunochemical detection of the allergens composition of spore and mycelium of C. cladosporioides was studied by electroblotting using sera from Cladosporium allergic patients and 125 I- anti- human IgE. A MW range of allergens between 16 to 88 KDa was identified. The most important with a MW of 16, 20,30, 39, 43, 50, 60 and 88 KDa. RESULTS: The allergenic composition of spore and mycelium looked very similar. However, partial or total inhibition of the serum with a conidial or mycelial extract demonstrated that the total concentration of allergens (particulary 20 and 60 KDa molecules) was higher in the conidium than in the mycelium. CONCLUSIONS: These results indicated that conidium and mycelium contained the same allergenic determinants but at different concentration in the two propagule. Results with 50 % inhibited sera demonstrated also that the total concentration of allergens was higher in the conidium than in the mycelium.


Asunto(s)
Alérgenos/inmunología , Antígenos Fúngicos/inmunología , Cladosporium/inmunología , Inmunoglobulina E/inmunología , Antígenos Fúngicos/aislamiento & purificación , Western Blotting , Cladosporium/fisiología , Cladosporium/ultraestructura , Colodión , Reacciones Cruzadas , Electroforesis en Gel de Poliacrilamida , Filtración , Proteínas Fúngicas/inmunología , Proteínas Fúngicas/aislamiento & purificación , Glicoproteínas/inmunología , Glicoproteínas/aislamiento & purificación , Humanos , Inmunoglobulina E/sangre , Peso Molecular , Micelio/inmunología , Especificidad de Órganos , Especificidad de la Especie , Esporas Fúngicas/inmunología , Extractos de Tejidos/inmunología
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