RESUMEN
TM0601p is a whey protein isolate derived from cow milk, containing a concentrated amount of transforming growth factor ß2 (TGF-ß2), and is intended for nutritional use in infants and adults. In vivo and in vitro studies have been performed to evaluate the safety of this product. In a 13-week toxicity study, treatment of adult Sprague-Dawley rats by gavage at up to 2000mg/kg/day did not result in any significant findings other than minor non-adverse changes in urinary parameters in females. The no-observed-adverse-effect level (NOAEL) was established as 2000mg/kg/day. In a juvenile toxicity study, rat pups received 600mg/kg/day by gavage from postnatal day (PND) 7 to PND 49. Transient lower bodyweight gain in the pre-weaning period was attributed to gastrointestinal effects of the viscous test material; following weaning, bodyweight gain was comparable to the vehicle controls. Reduced eosinophil counts and changes in urinary parameters (females) were recorded in treated pups at PND 49, and higher thymus weights were recorded in males only at the end of the recovery period (Day 77). None of the findings were considered adverse. There were no other significant findings and the NOAEL was established as 600mg/kg/day. No evidence of genotoxicity was seen in the bacterial reverse mutation test or the in vitro micronucleus test. Overall the results obtained present a reassuring safety profile for TM0601p.
Asunto(s)
Proteínas de la Leche/efectos adversos , Factor de Crecimiento Transformador beta2/efectos adversos , Animales , Femenino , Tracto Gastrointestinal/efectos de los fármacos , Masculino , Pruebas de Micronúcleos/métodos , Pruebas de Mutagenicidad , Nivel sin Efectos Adversos Observados , Tamaño de los Órganos/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Seguridad , Aumento de Peso/efectos de los fármacos , Proteína de Suero de LecheRESUMEN
Risk assessment describes the impact of a particular hazard as a function of dose and exposure. It forms the foundation of risk management and contributes to the overall decision-making process, but is not its endpoint. This paper outlines a risk analysis framework to underpin decision-making in the area of allergen cross-contact. Specifically, it identifies challenges relevant to each component of the risk analysis: risk assessment (data gaps and output interpretation); risk management (clear and realistic objectives); and risk communication (clear articulation of risk and benefit). Translation of the outputs from risk assessment models into risk management measures must be informed by a clear understanding of the model outputs and their limitations. This will lead to feasible and achievable risk management objectives, grounded in a level of risk accepted by the different stakeholders, thereby avoiding potential unintended detrimental consequences. Clear, consistent and trustworthy communications actively involving all stakeholders underpin these objectives. The conclusions, integrating the perspectives of different stakeholders, offer a vision where clear, science-based benchmarks form the basis of allergen management and labelling, cutting through the current confusion and uncertainty. Finally, the paper recognises that the proposed framework must be adaptable to new and emerging evidence.
Asunto(s)
Alérgenos/toxicidad , Alérgenos/administración & dosificación , Relación Dosis-Respuesta Inmunológica , HumanosRESUMEN
Biopesticides can be effective in controlling their target pest. However, research regarding allergenicity and asthma development is limited. We compared the ability of fungal biopesticide Metarhizium anisopliae (MACA) and house dust mite (HDM) extracts to induce allergic responses in BALB/c mice. The extracts were administered by intratracheal aspiration at doubling doses (2.5-80 µg protein) 4X over a four-week period. Three days after the last exposure, serum and bronchoalveolar lavage fluid (BALF) were collected. The extracts' relative allergenicity was evaluated based on response robustness (lowest significant dose response compared to control (0 µg)). MACA induced a more robust serum total IgE response than HDM. However, in the antigen-specific IgE assay, a similar dose of both MACA and HDM was required to achieve the same response level. Our data suggest a threshold dose of MACA for allergy induction and that M. anisopliae may be similar to HDM in allergy induction potential.
RESUMEN
A report by the Institute of Medicine suggested that more research is needed to better understand mold effects on allergic disease, particularly asthma development. The authors compared the ability of the fungus Stachybotrys chartarum (SCE) and house dust mite (HDM) extracts to induce allergic responses in BALB/c mice. The extracts were administered by intratracheal aspiration (IA) at several doses (0, 2.5, 5, 10, 20, 40, and 80 microg) 4 times over a 4-week period. Three days after the last IA exposure, serum and bronchoalveolar lavage fluid (BALF) were collected. The relative allergenicity of the extracts was evaluated based on the lowest dose that induced a significant response compared to control (0 microg) and the linear regression slope analysis across the dose range. SCE induced a more robust response than HDM for BALF some inflammatory cells (macrophage and neutrophils), whereas HDM induced more robust BALF lymphocyte and eosinophil responses. Although SCE induced a more robust serum total immunoglobulin E (IgE) response than did HDM, the induction of a similar response in a functional, antigen-specific IgE assay required approximately twice as much SCE as HDM. Even though SCE demonstrates the ability to induce allergic responses in the mouse model, considering the importance and relevance of eosinophil, lymphocyte, and antigen-specific IgE in allergic airway disease, it is concluded that HDM is more potent than SCE in the induction of allergic responses. These data suggest a threshold dose for SCE allergy induction. Furthermore, in damp water-damaged environments, exposure to S. chartarum might easily exceed the sensitization threshold for a susceptible population.
Asunto(s)
Antígenos Dermatofagoides/inmunología , Antígenos Fúngicos/inmunología , Hipersensibilidad/etiología , Pyroglyphidae/inmunología , Stachybotrys/inmunología , Animales , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Línea Celular Tumoral , Modelos Animales de Enfermedad , Femenino , Hipersensibilidad/sangre , Hipersensibilidad/inmunología , Inmunoglobulina E/sangre , L-Lactato Deshidrogenasa/análisis , Recuento de Leucocitos , Modelos Lineales , Ratones , Ratones Endogámicos BALB C , Péptido Hidrolasas/metabolismo , Ratas , beta-N-Acetilhexosaminidasas/análisisRESUMEN
BACKGROUND: Previous studies have demonstrated that Metarhizium anisopliae extract can induce responses characteristic of human allergic asthma in a mouse model. The study objectives were (1) to identify and characterize the M. anisopliae mycelia extract (MYC) proteins that are recognized by mouse serum IgE, (2) to determine if human serum IgE reacts with these proteins, and (3) to determine if these IgE-reactive proteins are found in other fungi. METHODS: Asthmatic human serum IgE, M. anisopliae crude antigen (MACA) immunized mouse serum IgE, and anti-catalase antibodies were used to probe one- and two-dimensional gel electrophoresis blots of MYC. RESULTS: Mass spectrometry analysis identified catalase as a mouse IgE-reactive protein. This identification was confirmed by assaying catalase activity in the extract and extract immunoblots probed with anti-catalase antibody. Six adult asthmatic sera contained IgE, but not IgG, that was reactive with mycelia extract proteins. A similar protein profile was seen when blots were probed with either mouse anti-MACA IgE or anti-bovine liver catalase antibodies. Furthermore, these mouse anti-MACA and anti-catalase antibodies were cross-reactive with other mold extracts (skin prick testing mix) and Aspergillus niger catalase. CONCLUSIONS: Some human asthmatics have developed IgE that reacts with an M. anisopliae catalase, most likely due to cross-reactivity (minimal IgG development). The cross-reactivity among fungal catalases suggests that IgE-reactive catalase might be useful for exposure assessment. Additionally, the similarity of protein profiles visualized with both human and mouse serum IgE suggests that allergy hazard identification can be facilitated using a mouse model.
Asunto(s)
Antígenos Fúngicos/metabolismo , Asma/inmunología , Catalasa/metabolismo , Inmunoglobulina E/metabolismo , Metarhizium/inmunología , Adulto , Animales , Antígenos Fúngicos/inmunología , Asma/sangre , Asma/microbiología , Catalasa/inmunología , Bovinos , Femenino , Humanos , Inmunización Secundaria , Inmunoglobulina E/inmunología , Metarhizium/enzimología , Ratones , Ratones Endogámicos BALB C , Micelio/metabolismo , Unión ProteicaRESUMEN
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 , VacunaciónRESUMEN
Neurotrophins, including nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin (NT)-3, have been implicated in the pathogenesis of many features and symptoms of asthma. The role of neurotrophins in fungal allergic asthma, however, is unknown. Repeated pulmonary challenge with Penicillium chrysogenum extract (PCE) induces dose-dependent allergic asthma-like responses in mice. The aim of this study was to investigate whether neurotrophins are involved in the PCE-induced allergic airway response in mice. Mice were exposed to 10, 20, 50, or 70 microg PCE by involuntary aspiration 4 times over 1 mo. Bronchial alveolar lavage fluid (BALF) was collected immediately before and after the final exposure. The levels of NGF, NT-3, and NT-4 were determined by enzyme-linked immunosorbent assay (ELISA). The lungs were fixed and processed for immunohistochemical examination of NGF production. PCE-exposed mice had dose-dependent increases in NGF, NT-3, and NT-4 in both BALF and sera. Exposures to PCE produced elevation in positive immunohistochemical staining for NGF in the airway epithelium and smooth muscle cells, in addition to infiltrated cells such as mononuclear cells, eosinophils, and macrophages. Taken together, this is the first study to link fungal allergic asthma in an experimental model with enhanced production of neurotrophins in the airways, and suggests that neurotrophins may play a role in the etiology of mold-induced asthma in humans.
Asunto(s)
Asma/inmunología , Asma/metabolismo , Factores de Crecimiento Nervioso/metabolismo , Penicillium chrysogenum/patogenicidad , Animales , Asma/microbiología , Líquido del Lavado Bronquioalveolar/química , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Femenino , Hipersensibilidad/inmunología , Inmunohistoquímica , Ratones , Ratones Endogámicos BALB CRESUMEN
Indoor mold has been associated with the development of allergic asthma. Penicillium chrysogenum, a common indoor mold, is known to have several allergens and can induce allergic responses in a mouse model of allergic penicilliosis. Our hypothesis is that soluble components of P. chrysogenum (PCE) can dose-dependently induce responses typical of allergic asthma in BALB/c mice. Mice were exposed to 10, 20, 50, or 70 microg of PCE by involuntary aspiration four times over a 4-week period. Serum and bronchoalveolar lavage fluid (BALF) were collected before (day 0), and at days 1 and 3 following the final exposure. PCE-exposed mice demonstrated dose-dependent increases in: BALF total cell numbers including eosinophil, serum and BALF total IgE levels, BALF IL-5 levels, and increased severity of histopathologic lesions. A single exposure to the highest dose of PCE resulted in edema and cellular damage but not immune responses. Four exposures to Metarhizium anisopliae crude antigen (10 microg, positive control) resulted in equivalent or greater allergic asthma-like responses than those demonstrated by multiple exposures to 50 or 70 microg of PCE. Multiple exposures to 70 microg of PCE showed increased allergen-triggered immediate respiratory responses as well as non-specific airway hyperresponsiveness to methacholine as assessed by barometric whole-body plethysmography. Taken together, repeated pulmonary challenge with P. chrysogenum extract induced dose-dependent allergic asthma-like responses in mice.
Asunto(s)
Alérgenos/inmunología , Hipersensibilidad/inmunología , Penicillium chrysogenum/inmunología , Alérgenos/administración & dosificación , Animales , Antígenos Fúngicos/administración & dosificación , Antígenos Fúngicos/inmunología , Asma/inmunología , Asma/metabolismo , Líquido del Lavado Bronquioalveolar/inmunología , Relación Dosis-Respuesta Inmunológica , Femenino , Humanos , Hipersensibilidad/metabolismo , Inmunoglobulina E/biosíntesis , Pulmón/inmunología , Pulmón/metabolismo , Cloruro de Metacolina/administración & dosificación , Ratones , Ratones Endogámicos BALB C , Penicillium chrysogenum/aislamiento & purificaciónRESUMEN
Inflammation plays a major role in the development of chronic diseases such as cardiovascular disease and Type 2 diabetes. Further, it was demonstrated that obese animals and humans have significantly higher levels of circulating proinflammatory cytokines, such as interleukin-6 (IL-6). The aim of this study was to determine whether adipose tissue could be a major source of circulating IL-6 in leptin-deficient obese (ob/ob) mice by comparing the expression of IL-6 in different tissues of ob/ob mice. Our secondary goal was to determine whether preadipocytes are the source of adipose tissue IL-6. The ob/ob mice had higher levels of plasma IL-6 (P < 0.05) and adipose tissue IL-6 mRNA (P < 0.05) compared with lean mice. Interestingly, IL-6 mRNA levels of liver and spleen were not different between ob/ob and lean mice, whereas adipose tissue IL-6 mRNA levels were higher in the ob/ob mice compared with lean mice (P < 0.05). In addition, we showed that IL-6 secretion from the adipose tissue stromal vascular fraction cells was higher than that from fully differentiated adipocytes (P < 0.001). We further demonstrated that 3T3-L1 preadipocytes had significantly higher levels of lipopolysaccharide (LPS)-stimulated IL-6 mRNA and IL-6 secretion than differentiated 3T3-L1 adipocytes. Taken together, these data suggest that adipose tissue and preadipocytes from the adipose tissue stromal vascular fraction may contribute significantly to the increased plasma IL-6 levels in ob/ob mice.
Asunto(s)
Adipocitos/metabolismo , Inflamación/metabolismo , Interleucina-6/metabolismo , Macrófagos Peritoneales/metabolismo , Obesidad/metabolismo , Células 3T3-L1 , Animales , Glucemia , Insulina/sangre , Leptina/sangre , Leptina/deficiencia , Lipopolisacáridos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones ObesosRESUMEN
Deoxynivalenol (DON, vomitoxin) is a trichothecene mycotoxin that potentially mediates toxicity by upregulating proinflammatory cytokine gene expression in vitro and in vivo. The purpose of this study was to test the hypothesis that DON-induced activation of mitogen-activated protein kinases (MAPKs) mediates transcriptional and posttranscriptional upregulation of TNF-alpha gene expression. RNAse protection assay revealed that DON at 100 to 500 ng/ml induced mRNA expression of TNF-alpha as well as IL-6, IFN-gamma, TGFbeta-1, and TGFbeta-3 and that these effects were potentiated by 100 ng/ml lipopolysaccharide (LPS). DON was found to induce phosphorylation of p38 kinase, extracellular signal-regulated kinases (ERKs), and c-Jun amino terminal kinases (JNKs) in a dose-dependent manner in the RAW 264.7 murine macrophage model. A luciferase reporter gene driven by the murine TNF-alpha promoter was used to assess the role of various MAPKs on DON upregulation of TNF-alpha gene transcription. The p38 inhibitor SB203580 reduced induction of luciferase activity by DON, LPS, and DON + LPS. In addition, the ERK inhibitor PD 98059 blocked DON- and DON + LPS-induced luciferase activity whereas the JNK inhibitor impaired LPS- and DON + LPS-induced luciferase activity. To study the effects of MAPKs on DON-induced TNF-alpha mRNA stability, an asynchronous model was used whereby cells were pretreated with LPS for 4 h and the medium was removed. Following incubation with medium containing a transcription inhibitor, 5,6-dichloro-beta-D-ribofuranosyl-benzimidazole, MAPK inhibitors and/or DON (250 ng/ml) cultures were monitored for TNF-alpha mRNA expression. DON-induced TNF-alpha mRNA stabilization was abrogated in the presence of SB 203580, whereas the stabilization by DON was not affected by PD 98059 or SP 600125. To verify the role of MAPKs in DON + LPS-induced TNF-alpha production, cells were incubated with LPS, DON, or LPS + DON for 18 h in the presence of inhibitors. ELISA of supernatant indicated that induction of TNF-alpha production by DON alone was significantly reduced by SB 203580 and PD 98059, whereas all three inhibitors blocked LPS- and DON + LPS-induced TNF-alpha production. Taken together, these results suggest that relative to DON-induced TNF-alpha mRNA expression, p38 and ERK activation contribute to DON-induced transcriptional upregulation whereas p38 plays a role in increasing mRNA stability.
Asunto(s)
Proteínas Quinasas Activadas por Mitógenos/biosíntesis , Procesamiento Postranscripcional del ARN/efectos de los fármacos , Activación Transcripcional , Tricotecenos/toxicidad , Factor de Necrosis Tumoral alfa/biosíntesis , Animales , Línea Celular , Combinación de Medicamentos , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/enzimología , Ratones , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , ARN Mensajero/metabolismo , Factor de Necrosis Tumoral alfa/genética , Regulación hacia Arriba/efectos de los fármacos , Proteínas Quinasas p38 Activadas por MitógenosRESUMEN
The trichothecenes are a group of mycotoxins that target leukocytes and have a wide range of immunomodulatory effects. Differential display analysis was applied to assess the effects of the trichothecenes deoxynivalenol (vomitoxin, DON) and satratoxin G (SG), on mRNA in the RAW 264.7 macrophage cell line. Cells were incubated with DON (1 microg/ml) or SG (5 ng/ml) for 2 h and total RNA then subjected to RT-PCR with a set of oligo(dT) primers. Resultant cDNA was amplified using an oligo (dT) downstream primer and an arbitrary decanucleotide upstream primer to make 35S-labeled PCR products. After separation of the products in denaturing polyacrylamide gel, 23 differentially expressed cDNA fragments were isolated and sequenced. Two of these were identified as known genes, namely, macrophage inflammatory protein-2 (MIP-2), a potent neutrophil chemoattractant involved in tissue injury and inflammation, and complement 3a receptor (C3aR), a proinflammatory mediator. Both MIP-2 and C3aR mRNAs were up-regulated by DON while only MIP-2 mRNA was induced by SG. Using commercially available antibodies, MIP-2 protein was also found to be induced by both DON and SG in RAW 264.7 cell cultures. When mice were treated with DON (12.5 mg/kg), splenic MIP-2 mRNA and serum MIP-2 levels were increased. MIP-2 mRNA and serum MIP-2 levels were synergistically increased when mice were co-treated with DON and LPS. Up-regulation of MIP-2 and C3aR are consistent with previous reports of trichothecene-induced inflammatory gene up-regulation and suggest that the specific genes affected may depend on trichothecene structures.
Asunto(s)
Antígeno de Macrófago-1/biosíntesis , Macrófagos/efectos de los fármacos , Monocinas/biosíntesis , Tricotecenos/farmacología , Animales , Quimiocina CXCL2 , Cruzamientos Genéticos , Perfilación de la Expresión Génica , Antígeno de Macrófago-1/genética , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Monocinas/genética , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tricotecenos/inmunología , Tricotecenos/metabolismo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
The satratoxins and other macrocyclic trichothecene mycotoxins are produced by Stachybotrys, a mold that is often found in water-damaged dwellings and office buildings. To test the potential immunomodulatory effects of these mycotoxins, RAW 264.7 murine macrophage cells were treated with various concentrations of satratoxin G (SG), isosatratoxin F (iSF), satratoxin H (SH), roridin A (RA), and verrucarin A (VA) for 48 h in the presence or absence of suboptimal concentra-tion of lipopolysaccharide (LPS, 50 ng/ml), and tumor necrosis factor-alpha (TNF-alpha ) and interleukin-6 (IL-6) production were assayed by enzyme-linked immunosorbent assay (ELISA). In LPS-stimulated cultures, TNF-alpha supernatant concentrations were significantly increased in the presence of 2.5, 2.5, and 1 ng/ml of SG, SH, and RA, respectively, whereas IL-6 concentrations were not affected by the same concentrations these macrocyclic trichothecenes. When cells that were treated with LPS and SG (2.5 ng/ml) were evaluated by real-time polymerase chain reaction (PCR),TNF-alpha mRNA was found to increase at 24, 36, and 48 h compared to control cells. At higher concentrations, cytokine production and cell viability were markedly impaired in LPS-stimulated cells. Without LPS stimulation, neither TNF-alpha, nor IL-6 was induced. These results indicate that low concentrations of macrocyclic trichothecenes superinduce expression of TNF-alpha, whereas higher concentrations of these toxins are cytotoxic and concurrently reduce cytokine production. The capacity of satratoxins and other macrocyclic trichothecenes to alter cytokine production may play an etiologic role in outbreaks of Stachybotrys-associated human illnesses.
Asunto(s)
Interleucina-6/análisis , Lipopolisacáridos/efectos adversos , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Tricotecenos/toxicidad , Factor de Necrosis Tumoral alfa/análisis , Análisis de Varianza , Animales , Línea Celular , Ensayo de Inmunoadsorción Enzimática , Humanos , Inflamación/inmunología , Interleucina-6/inmunología , Lipopolisacáridos/inmunología , Enfermedades Pulmonares Fúngicas/inmunología , Enfermedades Pulmonares Fúngicas/microbiología , Activación de Macrófagos/inmunología , Ratones , Reacción en Cadena de la Polimerasa , Stachybotrys , Tricotecenos/química , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Although satratoxin G (SG) is the primary macrocyclic trichothecene produced by Stachybotrys chartarum, a fungus frequently associated with outbreaks of indoor air illness, little is known about the actual quantities of this mycotoxin in suspect or normal building environments. Using antibodies produced against an SG hemisuccinate-bovine serum albumin conjugate, we devised an enzyme-linked immunosorbent assay that enabled the detection of as little as 100 pg/mL of SG and that could also detect other members of the satratoxin family. Detection by this assay of satratoxins in methanolic extracts of S. chartarum in rice cultures was comparable to that of high-performance liquid chromatography, indicating that this assay will be useful in detecting SG and other satratoxins in environmental samples.