Novel acute hypersensitivity pneumonitis model induced by airway mycosis and high dose lipopolysaccharide.

BACKGROUND: Inhalation of fungal spores is a strong risk factor for severe asthma and experimentally leads to development of airway mycosis and asthma-like disease in mice. However, in addition to fungal spores, humans are simultaneously exposed to other inflammatory agents such as lipopolysaccharide (LPS), with uncertain relevance to disease expression. To determine how high dose inhalation of LPS influences the expression of allergic airway disease induced by the allergenic mold Aspergillus niger (A. niger). METHODS: C57BL/6J mice were intranasally challenged with the viable spores of A. niger with and without 1 µg of LPS over two weeks. Changes in airway hyperreactivity, airway and lung inflammatory cell recruitment, antigen-specific immunoglobulins, and histopathology were determined. RESULTS: In comparison to mice challenged only with A. niger, addition of LPS (1 µg) to A. niger abrogated airway hyperresponsiveness and strongly attenuated airway eosinophilia, PAS+ goblet cells and TH2 responses while enhancing TH1 and TH17 cell recruitment to lung. Addition of LPS resulted in more severe, diffuse lung inflammation with scattered, loosely-formed parenchymal granulomas, but failed to alter fungus-induced IgE and IgG antibodies. CONCLUSIONS: In contrast to the strongly allergic lung phenotype induced by fungal spores alone, addition of a relatively high dose of LPS abrogates asthma-like features, replacing them with a phenotype more consistent with acute hypersensitivity pneumonitis (HP). These findings extend the already established link between airway mycosis and asthma to HP and describe a robust model for further dissecting the pathophysiology of HP.

Primary Cutaneous Aspergillosis in Immunocompetent Patients Due to Aspergillus niger: A Report of 4 Cases.

ABSTRACT: Primary cutaneous aspergillosis is a cutaneous fungal infection due to the direct inoculation of spores of Aspergillus species into the disrupted skin. Primary cutaneous aspergillosis presents with a variety of localized cutaneous lesions, such as erythematous macules, papules, plaques, or nodules that can progress to necrosis, erosion, ulceration, or fistulization. Many species of Aspergillus can cause the disease, and one of them is Aspergillus niger that rarely affects immunocompetent patients and that has peculiar characteristics on the histopathological examination. We present a series of 4 cases of immunologically competent patients presenting with primary cutaneous aspergillosis caused by A. niger.

Activation of cellular immune response in insect model host Galleria mellonella by fungal α-1,3-glucan.

Alpha-1,3-glucan, in addition to ß-1,3-glucan, is an important polysaccharide component of fungal cell walls. It is reported for many fungal species, including human pathogenic genera: Aspergillus, Blastomyces, Coccidioides, Cryptococcus, Histoplasma and Pneumocystis, plant pathogens, e.g. Magnaporthe oryzae and entomopathogens, e.g. Metarhizium acridum. In human and plant pathogenic fungi, α-1,3-glucan is considered as a shield for the ß-1,3-glucan layer preventing recognition of the pathogen by the host. However, its role in induction of immune response is not clear. In the present study, the cellular immune response of the greater wax moth Galleria mellonella to Aspergillus niger α-1,3-glucan was investigated for the first time. The changes detected in the total hemocyte count (THC) and differential hemocyte count (DHC), formation of hemocyte aggregates and changes in apolipophorin III localization indicated activation of G. mellonella cellular mechanisms in response to immunization with A. niger α-1,3-glucan. Our results, which have clearly demonstrated the response of the insect immune system to this fungal cell wall component, will help in understanding the α-1,3-glucan role in immune response against fungal pathogens not only in insects but also in mammals, including humans.

Seroprevalence of Aspergillus IgG and disease prevalence of chronic pulmonary aspergillosis in a country with intermediate burden of tuberculosis: a prospective observational study.

OBJECTIVES: Chronic pulmonary aspergillosis (CPA) is an emerging global disease with tuberculosis (TB) being the most important risk factor. Epidemiologic data on the seroprevalence of Aspergillus IgG and prevalence of CPA in different areas, especially in country with intermediate burden of TB, are lacking. METHODS: We prospectively recruited healthy volunteers, TB close contacts, active TB patients and participants with old pulmonary TB in Taiwan during 2012-2019. We measured serum Aspergillus fumigatus and niger-specific IgG levels and assessed if the participants were having CPA. RESULTS: A total of 1242 participants (including 200 healthy volunteers, 326 TB close contacts, 524 active TB patients and 192 old TB cases) were recruited. Using 27 mgA/L (milligrams of antigen-specific antibodies per liter) as cut-off level, the seropositive rate of A. fumigatus-specific IgG was 33.0% (66/200), 37.7% (123/326), 26.5% (139/524) and 43.2% (83/192) among the four groups, respectively. In multivariate logistic regression, pulmonary cavitation (OR 1.73; 95% CI 1.07-2.80), female sex (OR 1.49; 95% CI 1.14-1.95), old TB (OR 1.59; 1.05-2.42) were independent risk factors for Aspergillus IgG positivity. One (0.2%) active TB patient and four (2.1%) old TB patients developed CPA. Correlation between A. fumigatus and A. niger-specific IgG was high (Spearman correlation coefficient: 0.942). DISCUSSION: Geographic variation in Aspergillus IgG seroprevalence and CPA prevalence exists. A universal cut-off value for Aspergillus IgG may not exist. In areas and populations in which background Aspergillus IgG level is unknown, Aspergillus IgG may be better used as a test of exclusion for CPA using prespecified cut-off level.

Aspergillus fumigatus establishes infection in zebrafish by germination of phagocytized conidia, while Aspergillus niger relies on extracellular germination.

Among opportunistically pathogenic filamentous fungi of the Aspergillus genus, Aspergillus fumigatus stands out as a drastically more prevalent cause of infection than others. Utilizing the zebrafish embryo model, we applied a combination of non-invasive real-time imaging and genetic approaches to compare the infectious development of A. fumigatus with that of the less pathogenic A. niger. We found that both species evoke similar immune cell migratory responses, but A. fumigatus is more efficiently phagocytized than A. niger. Though efficiently phagocytized, A. fumigatus conidia retains the ability to germinate and form hyphae from inside macrophages leading to serious infection even at relatively low infectious burdens. By contrast, A. niger appears to rely on extracellular germination, and rapid hyphal growth to establish infection. Despite these differences in the mechanism of infection between the species, galactofuranose mutant strains of both A. fumigatus and A. niger display attenuated pathogenesis. However, deficiency in this cell wall component has a stronger impact on A. niger, which is dependent on rapid extracellular hyphal growth. In conclusion, we uncover differences in the interaction of the two fungal species with innate immune cells, noticeable from very early stages of infection, which drive a divergence in their route to establishing infections.

In vitro immunostimulatory potential of fungal ß-glucans in pacific red snapper (Lutjanus peru) cells.

This study attempts to describe the immunostimulatory effects of three fungal glucans on innate immunity responses in an in vitro assays using Pacific red snapper leukocytes. First, the yield glucans obtained was higher in Aspergillus niger, follow by Aspergillus ochraceus and Alternaria botrytis (40, 20 and 10%, respectively). Structural characterization of these fungal glucans by proton nuclear magnetic resonance (NMR) indicated structures containing (1-6)-branched (1-3)-ß-D-glucan. The immunostimulatory activity of fungal glucans were assessed in head-kidney leukocytes at 24 h using colorimetric assays and molecular gene expression. In addition, the response against bacterial infection using Aeromonas hydrophila was evaluated by flow cytometry with annexin V/propidium iodide. Leukocytes responded positively to fungal glucans where the viability was higher than 80%. Interestingly, A. niger ß-glucans enhanced the phagocytic ability and capacity in head-kidney leukocytes. Immunological assays reveled an increased in nitric oxide production, myeloperoxidase, superoxide dismutase and catalase activities, in fish stimulated with A. niger ß-glucans. Induction of cytokines (IL-1ß, TNF-α, IL-6, IL-8 and IL-12) were more pronounced in A. niger ß-glucans leukocytes stimulated compared to other group. Finally, flow cytometry assay showed that A. botrytis and A. niger ß-glucans were able to inhibit apoptosis caused by Aeromonas hydrophila in the Pacific red snapper leukocytes indicating an immunostimulant potent response by fungi derived-glucans. These results strongly support the idea that fungal ß-glucans can stimulate the immune mechanism in head-kidney leukocytes and that Aspergillus niger ß-glucan possess immunostimulatory properties cell increasing viability, and reducing necrotic cell death caused by Aeromonas hydrophila.

Application of silica nanoparticles in maize to enhance fungal resistance.

In this study, maize treated with nanosilica (20-40 nm) is screened for resistance against phytopathogens such as Fusarium oxysporum and Aspergillus niger and compared with that of bulk silica. The resistivity is measured for disease index and expression of plant responsive compounds such as total phenols, phenylalanine ammonia lyase, peroxidase and polyphenol oxidase. The results indicate that nanosilica-treated plant shows a higher expression of phenolic compounds (2056 and 743 mg/ml) and a lower expression of stress-responsive enzymes against both the fungi. Maize expresses more resistance to Aspergillus spp., than Fusarium spp. These results show significantly higher resistance in maize treated with nanosilica than with bulk, especially at 10 and 15 kg/ha. In addition, hydrophobic potential and silica accumulation percentage of nanosilica treated maize (86.18° and 19.14%) are higher than bulk silica treatment. Hence, silica nanoparticles can be used as an alternative potent antifungal agent against phytopathogens.

Thymosin α1 activates complement receptor-mediated phagocytosis in human monocyte-derived macrophages.

Thymosin α1 (Tα1) is a naturally occurring thymic peptide used worldwide in clinical trials for the treatment of infectious diseases and cancer. The immunomodulatory activity of Tα1 on innate immunity effector cells has been extensively described, but its mechanism of action is not completely understood. We report that Tα1-exposed human monocyte-derived macrophages (MDMs) assume the typical activated morphology also exhibited by lipopolysaccharide-activated MDMs, but show a comparatively higher ability of internalizing fluorescent beads and zymosan particles. Tα1 exposure also promptly and dramatically stimulates MDM phagocytosis and killing of Aspergillus niger conidia starting as soon as 30 min after challenge. The effect is dose dependent and early coupled to low transcription of the proinflammatory cytokines tumor necrosis factor α and interleukin-6 and unmodified Toll-like receptor expression. The Tα1-stimulated phagocytosis is strictly dependent on the integrity of the microtubule network and protein kinase C activity and occurs by a variation in the classic zipper model, with recruitment of vinculin and actin at the phagosome exhibiting a punctate distribution. These findings indicate that, in human mature MDMs, Tα1 implements pathogen internalization and killing via the stimulation of the complement receptor-mediated phagocytosis. Our observations document that Tα1 is an early and potent activator of innate immunity and reinforce the concept of its pleiotropy.

CD11a polymorphisms regulate TH2 cell homing and TH2-related disease.

BACKGROUND: TH2-dependent diseases vary in severity according to genotype, but relevant gene polymorphisms remain largely unknown. The integrin CD11a is a critical determinant of allergic responses, and allelic variants of this gene might influence allergic phenotypes. OBJECTIVE: We sought to determine major CD11a allelic variants in mice and human subjects and their importance to allergic disease expression. METHODS: We sequenced mouse CD11a alleles from C57BL/6 and BALB/c strains to identify major polymorphisms; human CD11a single nucleotide polymorphisms were compared with allergic disease phenotypes as part of the international HapMap project. Mice on a BALB/c or C57BL/6 background and congenic for the other strain's CD11a allele were created to determine the importance of mouse CD11a polymorphisms in vivo and in vitro. RESULTS: Compared with the C57BL/6 allele, the BALB/c CD11a allele contained a nonsynonymous change from asparagine to aspartic acid within the metal ion binding domain. In general, the BALB/c CD11a allele enhanced and the C57BL/6 CD11a allele suppressed TH2 cell-dependent disease caused by the parasite Leishmania major and allergic lung disease caused by the fungus Aspergillus niger. Relative to the C57BL/6 CD11a allele, the BALB/c CD11a allele conferred both greater T-cell adhesion to CD54 in vitro and enhanced TH2 cell homing to lungs in vivo. We further identified a human CD11a polymorphism that significantly associated with atopic disease and relevant allergic indices. CONCLUSIONS: Polymorphisms in CD11a critically influence TH2 cell homing and diverse TH2-dependent immunopathologic states in mice and potentially influence the expression of human allergic disease.

Cleavage of fibrinogen by proteinases elicits allergic responses through Toll-like receptor 4.

Proteinases and the innate immune receptor Toll-like receptor 4 (TLR4) are essential for expression of allergic inflammation and diseases such as asthma. A mechanism that links these inflammatory mediators is essential for explaining the fundamental basis of allergic disease but has been elusive. Here, we demonstrate that TLR4 is activated by airway proteinase activity to initiate both allergic airway disease and antifungal immunity. These outcomes were induced by proteinase cleavage of the clotting protein fibrinogen, yielding fibrinogen cleavage products that acted as TLR4 ligands on airway epithelial cells and macrophages. Thus, allergic airway inflammation represents an antifungal defensive strategy that is driven by fibrinogen cleavage and TLR4 activation. These findings clarify the molecular basis of allergic disease and suggest new therapeutic strategies.

Sensibilisation of asthmatic patients to extracted antigens from strains of Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger.

OBJECTIVE: The main purpose of this study sought to evaluate the frequency of sensitivity of Iranian asthmatic patients to three regional Aspergillus species of fumigatus, flavus and niger, by detection of antigen-specific IgE in the patients' sera. PATIENTS AND METHODS: Crude extracts were prepared following the disruption of fungi cell walls by the application of glass beads and their protein fractions were isolated by SDS-PAGE. After electrotransfer of protein bands into the nitrocellulose membrane, IgE-immunoblotting was performed against the sera from 32 asthmatic patients in addition to 20 healthy controls. RESULTS: Our results interestingly showed that all of the studied Iranian asthmatic patients were sensitive to A. fumigatus and A. flavus antigens. This frequency was 65.6% in the case of A. niger, however, all control samples were negative. Age/sex analysis generally indicated higher sensitivities of young patients (<30 years old) to Aspergillus species with a statistical significance in the case of A. niger (P=0.02) and additionally more sensitivity of females. Using Immunoblotting assay, 23 IgE-reactive allergenic components from A. fumigatus, 15 from A. flavus and 13 from A. niger in a broad molecular weight spectrum were identified, among which several fragments were not previously reported. CONCLUSION: Overall, this study found a high frequency of sensitivity of Iranian asthmatic patients to regional isolates of A. fumigatus, A. flavus and A. niger, which suggested the importance of these species in development of asthma. Moreover, we reported allergenic profiles of Iranian isolates in different patterns not previously observed.

[Pathogenesis of invasive fungal infections]. / Patogenia de la infección fúngica invasora.

Invasive fungal infections remain a life-threatening disease. The development of invasive fungal disease is dependent on multiple factors, such us colonization and efficient host immune response. We aimed to review the pathogenesis of invasive fungal infections, in particular, those caused by Candida and Aspergillus. For this we propose, to describe the fungal characteristics, to detail the host defence mechanisms against fungus and to analyse the host risk factors for invasive fungal infection.

Hypersensitivity testing for Aspergillus fumigatus IgE is significantly more sensitive than testing for Aspergillus niger IgE.

We sought to determine if sufficient redundancy exists between specific IgE testing for Aspergillus fumigatus and Aspergillus niger to eliminate one of the assays in determining Aspergillus hypersensitivity. We reviewed regional laboratory results comparing A fumigatus-specific IgE with A niger-specific IgE using the Pharmacia UniCAP system (Pharmacia, Kalamazoo, MI). By using the Fisher exact test as an index of concordance among paired results, we showed a significant difference between 109 paired samples for the presence of specific IgE to A fumigatus and A niger (P < .0001). Of these specimens, 94 were negative for IgE to both species, 10 were positive for A fumigatus and negative for A niger; no specimen was positive for A niger and negative for A fumigatus. We conclude that A fumigatus-specific IgE is sufficient to detect Aspergillus hypersensitivity. The assay for A niger-specific IgE is redundant, less sensitive, and unnecessary if the assay for specific IgE for A fumigatus is performed.

Characterization of blood beta-1,3-glucan and anti-beta-glucan antibody in hemodialysis patients using culinary-medicinal Royal Sun Agaricus, Agaricus brasiliensis S. Wasser et al. (Agaricomycetideae).

Beta-glucan is a major component of fungal cell walls and shows various immunopharmacological activities including antitumor activity. Previously, we detected anti-beta-glucan antibody in human sera. Anti-beta-glucan antibody participates in the immune response to fungal cell wall beta-glucan. Patients on dialysis are at high risk of infection including fungal infections. We examined the plasma beta-glucan level and the titer of anti-beta-glucan antibody in dialysis patients. We measured plasma beta-1,3-glucan concentrations with the limulus G test and anti-beta-glucan antibody titers by ELISA with Candida beta-glucan-coated plates. We also examined the influence of the period of dialysis and the kind of dialysis membrane. The patients were positive for beta-1,3-glucan in their plasma. The anti-beta-glucan antibody titer was lower in the dialysis patients than in healthy volunteers. Long-term dialysis patients showed lower anti-beta-glucan antibody titers than short-term dialysis patients. No significant difference was found between the kinds of dialysis membrane. The titer of anti-beta-glucan antibody as recognition molecule of beta-glucan was low in dialysis patients compared with healthy volunteers. This is likely to be one factor explaining the sensitivity to infection of the dialysis patients. An appropriate application of culinary-medicinal mushroom such as Agaricus brasiliensis has potential for the prevention of fungal infection in dialysis patients.

Physico-chemical and clinico-immunologic studies on the allergenic significance of Aspergillus tamarii, a common airborne fungus.

Aspergillus-derived inhalant allergens play an important role in the etiology of allergic respiratory diseases. In the present study, we investigated the allergenic potential of Aspergillus tamarii, quantified its airborne content, identified its major/minor allergens, evaluated heterogeneity of patients' IgE response to its allergens and cross-reactivity of its allergens with other Aspergillus allergens. Skin prick tests with A tamarii extract were performed on 300 patients of bronchial asthma/allergic rhinitis and 20 healthy volunteers. Sixty-six patients (22%) elicited positive cutaneous reactions to A tamarii extract. Only one of the 20 non-allergic healthy volunteer showed a mild positive cutaneous reaction. Allergen-specific IgE levels increased with increase in patients' cutaneous response (0% in negative to 100% in 3+/4+). The skin positivity and allergen-specific IgE levels were significantly higher in patients compared to healthy volunteers (P>0.05). However, no differences were found for these two parameters among patients of bronchial asthma, allergic rhinitis and bronchial asthma with allergic rhinitis. The airborne A tamarii allergen content was highest in February and October. A tamarii extract revealed at least 22 proteins (13.3-120 kDa). Seventeen of these proteins bound patients' IgE with six being major allergens (13.3, 23, 25, 34, 39.5, 43 kDa). Three major allergens (13.3, 34, 43 kDa) were found to cross-react with A flavus and one (34 kDa) with A niger. Our results revealed that A tamarii allergen(s) are present in the air, which might serve as important inhalant allergens in IgE-mediated allergic respiratory diseases.

Influencia de la concentración de inóculo en la producción de celulasa y xilanasa por aspergillus niger / Influence of inoculum concentration on the cellulase and xylanase production by aspergillus niger

Existe un gran interés por el uso de enzimas lignocelulolíticas en varias industrias, y en la biodegradación de biomasa para la producción de biocombustibles y otras aplicaciones. Entre las fuentes microbianas de enzimas, Aspergillus niger es uno de los microorganismos más utilizados en la producción de enzimas industriales, debido a sus niveles altos de secreción de proteína y a su condición GRAS (generally regarded as safe). El objetivo del presente estudio fue evaluar la influencia de la concentración de inóculo en la morfología y producción de celulasas y xilanasas con A. niger en cultivo sumergido. Para ello, fueron inoculados matraces de 250 mL con 40 mL de medio con 3% (v/v) de una suspensión de 104 o 108 esporas por mililitro e incubados a 28 ºC y 175 rpm durante 120 horas. Se utilizaron 10 g*L-1 de lactosa como fuente de carbono. En cada caso se determinó la cantidad de biomasa, la proteína extracelular soluble, lactosa residual, actividad celulasa total y xilanasa cada 24 horas. Aunque no hubo un efecto notorio en la morfología de crecimiento, salvo en el color y el diámetro de pellets obtenidos, sí se afectó la µmax (0,06 y 0,03 h-1 para 104 y 108 esporas*mL-1, respectivamente) y la concentración máxima de biomasa. Además, mientras que las productividades volumétricas de celulasa (ΓFPA) (8,2 y 8,0 UI.*L-1*h-1 para 104 y 108 esporas*mL-1, respectivamente) fueron similares para ambos inóculos, la productividad de xilanasa (ΓXIL) fue mayor para el inóculo más concentrado (29,7 y 33,4 UI¨*L-1*h-1 para 104 y 108 esporas*mL-1, respectivamente). Los resultados indican que la productividad de celulasas y xilanasas está estrechamente relacionada con la concentración de inóculo.

There is a great interest for the use of lignocellulolytic enzymes in several industries and in biomass degradation for production of biofuels and other applications. Among the microbial sources of enzymes, Aspergillus niger is one of the most used microorganisms in the production of industrial enzymes due to its high levels of protein secretion and its GRAS (generally regarded as safe) condition. The aim of the present study was to evaluate the influence of A. niger inoculum concentration in the morphology and production of cellulases and xylanases in submerged cultures. For this, 250 mL flasks containing 40 mL culture medium were inoculated with a 3% (v/v) of either 104 or 108 spores per milliliter suspension and incubated at 28 º C and 175 rpm during 120 hours. Lactose (10 g*L-1) was used as the carbon source. In each case, the amount of biomass, the extracellular soluble protein, residual lactose, total celullase activity and xylanase activity were determined every 24 hours. Even thought there was not a notorious effect on the growth morphology, except in color and diameter of pellets; µmax was affected (0.06 and 0.03 h-1 for 104 and 108 spores*mL-1, respectively) as well as maximum biomass concentration. In addition, while the volumetric productivity of cellulase (8.2 and 8.0 UI*L-1*h-1 for 104 and 108 spores*mL-1, respectively) were similar for both inocula, the productivity of xylanase was greater for the more concentrated inoculum (29.7 and 33.4 UI*L-1*h-1 for 104 and 108 spores*mL-1, respectively).The results show that cellulase and xylanase productivities are closely related to the inoculum concentration.

Efeito inibitório in vitro de extrato vegetal de Allium sativum sobre Aspergillus niger Tiegh / In vitro inhibitory effect of Allium sativum extract on Aspergillus niger Tiegh

O sisal (Agave sisalana Perrine ex Engelm.) é a principal fonte de extração de fibras duras vegetais do mundo. No Brasil, o cultivo ocupa extensa área de solos pobres na região semi-árida dos Estados da Bahia, Paraíba e Rio Grande do Norte, em regiões com escassa ou nenhuma alternativa para exploração de outras culturas. Tendo em vista a propriedade inibitória dos extratos vegetais sobre o desenvolvimento de fungos, avaliou-se o efeito inibitório do extrato vegetal de alho (Allium sativum L.) sobre o fungo Aspergillus niger isolados da cultura do sisal. O extrato vegetal foi incorporado ao meio BDA fundente, aproximadamente 45ºC, nas concentrações 0, 500, 1000, 2000, 50000, 10000 e 50000 mg L-1. Os resultados demonstraram que a dose de 50000 mg L-1 de alho foi eficiente na inibição do crescimento do fungo Aspergillus niger "extraídos" da cultura do sisal.

Sisal (Agave sisalana Perrine ex Engelm.) is the main source of plant hard fiber extraction in the world. In Brazil, its cultivation occupies a large area of poor soils in the semi-arid region of Bahia, Paraiba and Rio Grande do Norte States, and in regions with scarce or no alternative to explore other crops. Based on the inhibitory property of plant extracts on fungal development, the inhibitory effect of garlic (Allium sativum L.) extract was evaluated on the fungus Aspergillus niger isolated from sisal crop. The plant extract was incorporated into melting PDA medium around 45ºC, at the following concentrations: 0, 500, 1000, 2000, 50000, 10000, and 50000 mg L-1. (OBS: M. Elena, há duas doses repetidas; creio que a primeira seja 5000). The concentration 50000 mg L-1 garlic extract was effective in inhibiting the growth of the fungus Aspergillus niger extracted from sisal crop.

Toward a comprehensive understanding of allergic lung disease.

Allergic asthma is a respiratory disease induced by exposure to environmental agents that elicit allergic inflammation and transient airway obstruction and which produce the characteristic symptoms of cough and dyspnea. Prior to the advent of experimental models, asthma was believed to be caused primarily by the degranulation of mast cells and eosinophils primed by antigen-specific immunoglobulin E (IgE). More recent studies in mice have shown that T cells primarily mediate antigen-dependent airway obstruction and allergic inflammation through secretion of the cytokines interleukin 4 (IL- 4) and IL-13. Our additional studies indicate that a major environmental link to asthma may be through exposure to environmental proteinases and especially airway infection by proteinase-producing organisms such as fungi. Pending verification in humans, these findings suggest entirely new therapeutic interventions in asthma that include the restricted use of anti-inflammatory therapy and universal application of anti-fungal agents.