Fungi and allergic lower respiratory tract diseases.

Asthma is a common disorder that in 2009 afflicted 8.2% of adults and children, 24.6 million persons, in the United States. In patients with moderate and severe persistent asthma, there is significantly increased morbidity, use of health care support, and health care costs. Epidemiologic studies in the United States and Europe have associated mold sensitivity, particularly to Alternaria alternata and Cladosporium herbarum, with the development, persistence, and severity of asthma. In addition, sensitivity to Aspergillus fumigatus has been associated with severe persistent asthma in adults. Allergic bronchopulmonary aspergillosis (ABPA) is caused by A fumigatus and is characterized by exacerbations of asthma, recurrent transient chest radiographic infiltrates, coughing up thick mucus plugs, peripheral and pulmonary eosinophilia, and increased total serum IgE and fungus-specific IgE levels, especially during exacerbation. The airways appear to be chronically or intermittently colonized by A fumigatus in patients with ABPA. ABPA is the most common form of allergic bronchopulmonary mycosis (ABPM); other fungi, including Candida, Penicillium, and Curvularia species, are implicated. The characteristics of ABPM include severe asthma, eosinophilia, markedly increased total IgE and specific IgE levels, bronchiectasis, and mold colonization of the airways. The term severe asthma associated with fungal sensitization (SAFS) has been coined to illustrate the high rate of fungal sensitivity in patients with persistent severe asthma and improvement with antifungal treatment. The immunopathology of ABPA, ABPM, and SAFS is incompletely understood. Genetic risks identified in patients with ABPA include HLA association and certain T(H)2-prominent and cystic fibrosis variants, but these have not been studied in patients with ABPM and SAFS. Oral corticosteroid and antifungal therapies appear to be partially successful in patients with ABPA. However, the role of antifungal and immunomodulating therapies in patients with ABPA, ABPM, and SAFS requires additional larger studies.

Analysis of available diagnostic tests for latex sensitization in an at-risk population.

BACKGROUND: Lack of a Food and Drug Administration (FDA)-approved skin testing reagent for latex allergy in the United States requires reliance on patient history and serologic assays for diagnosis. OBJECTIVE: To determine the diagnostic sensitivity, specificity, and predictive values of an FDA-cleared antilatex IgE serology test and an enzyme-linked immunosorbent assay (ELISA) with various sources of latex protein antigens in an at-risk but unselected population of health care workers. METHODS: Health care workers underwent duplicate latex and serologic testing for latex specific IgE with the CAP assay and ELISA from June 1, 1998, through December 31, 2002. Logistic regression with receiver operating characteristic curve analysis determined the values, resulting in 98% and 99% specificity for the CAP assay and ELISA, respectively. RESULTS: Results of paired skin and serologic tests were available for 792 participants. Forty duplicate skin test results (5%) were positive. For the CAP assay, sensitivity was 35%; specificity, 98%; positive predictive value, 48.3%; and negative predictive value, 96.6%. ELISA demonstrated similar results. Multivariable logistic regression yielding a 98% or 99% specificity for the various ELISAs demonstrated that the adjusted odds of a positive skin test result significantly increased with positive CAP assay and ELISA results using a powdered glove extract. CONCLUSIONS: The performance of the FDA-cleared antilatex IgE serologic test for latex allergy has much lower sensitivity than previously reported. This finding confirms that this serologic test should be used only for patients with a history of latex allergy and not for screening the population with a low prevalence of latex sensitization.

Immune response among patients exposed to molds.

Macrocyclic trichothecenes, mycotoxins produced by Stachybotrys chartarum, have been implicated in adverse reactions in individuals exposed to mold-contaminated environments. Cellular and humoral immune responses and the presence of trichothecenes were evaluated in patients with mold-related health complaints. Patients underwent history, physical examination, skin prick/puncture tests with mold extracts, immunological evaluations and their sera were analyzed for trichothecenes. T-cell proliferation, macrocyclic trichothecenes, and mold specific IgG and IgA levels were not significantly different than controls; however 70% of the patients had positive skin tests to molds. Thus, IgE mediated or other non-immune mechanisms could be the cause of their symptoms.

Immune response modulation by curcumin in a latex allergy model.

BACKGROUND: There has been a worldwide increase in allergy and asthma over the last few decades, particularly in industrially developed nations. This resulted in a renewed interest to understand the pathogenesis of allergy in recent years. The progress made in the pathogenesis of allergic disease has led to the exploration of novel alternative therapies, which include herbal medicines as well. Curcumin, present in turmeric, a frequently used spice in Asia has been shown to have anti-allergic and inflammatory potential. METHODS: We used a murine model of latex allergy to investigate the role of curcumin as an immunomodulator. BALB/c mice were exposed to latex allergens and developed latex allergy with a Th2 type of immune response. These animals were treated with curcumin and the immunological and inflammatory responses were evaluated. RESULTS: Animals exposed to latex showed enhanced serum IgE, latex specific IgG1, IL-4, IL-5, IL-13, eosinophils and inflammation in the lungs. Intragastric treatment of latex-sensitized mice with curcumin demonstrated a diminished Th2 response with a concurrent reduction in lung inflammation. Eosinophilia in curcumin-treated mice was markedly reduced, co-stimulatory molecule expression (CD80, CD86, and OX40L) on antigen-presenting cells was decreased, and expression of MMP-9, OAT, and TSLP genes was also attenuated. CONCLUSION: These results suggest that curcumin has potential therapeutic value for controlling allergic responses resulting from exposure to allergens.

Specific antibodies to recombinant allergens of Aspergillus fumigatus in cystic fibrosis patients with ABPA.

BACKGROUND: Aspergillus fumigatus, a widely distributed fungus, has been implicated in causing life threatening infections as well as severe asthma and allergic diseases in man. Allergic affliction like allergic bronchopulmonary aspergillosis (ABPA) is a disabling lung disease frequently seen in patients with asthma and cystic fibrosis. Immunodiagnosis of the former is comparatively easier due to the availability of purified antigens and sensitive methods. However, this is not true with cystic fibrosis patients where the prevalence of ABPA is fairly high and the morbidity and mortality are significant. METHODS: In the present study, we have evaluated purified recombinant allergens from A. fumigatus, namely Asp f 1, f 2, f 3, f 4, and f 6 using ELISA and a semi-automated method (ImmunoCAP). We studied 17 patients each from cystic fibrosis with ABPA, and cystic fibrosis with asthma, 22 cystic fibrosis with no ABPA or asthma, and 11 age matched controls. RESULTS: The results indicate that no antigen, antibody or method is capable of differentiating cystic fibrosis (CF) with ABPA from other CF patients, although some allergens showed strong reaction or showed more prevalence among the patients studied. CONCLUSION: When results of several allergens such as Asp f 1, f 2, f 3, f 4, and f 6 in their binding to IgA, IgG, and IgE antibodies were analyzed, a more strong discrimination of CF patients with ABPA was possible from the other groups studied.

Hypersensitivity pneumonitis.

Hypersensitivity pneumonitis (HP) or extrinsic allergic alveolitis is a non-IgE mediated hypersensitivity disease initiated by inhalation and subsequent sensitisation to organic antigens. These diseases have been described in different occupational groups and present in acute, subacute or chronic forms based on the exposure to antigens and host response. Clinical features are dependent upon the stage of the disease and can include fever, chills, cough, dyspnoea, and weight loss. The immunopathogenesis involves both cellular immunity and antibody responses to inhaled antigens. Antibody response to the implicated antigen can be demonstrated in HP patients, but such antibodies are also detected in antigen exposed asymptomatic individuals. Bronchoalveolar lavage demonstrates lymphocytosis and preponderance of CD8+ cells. Pulmonary function studies demonstrate a restrictive pattern with diffusion defects. The diagnosis is difficult as no single test is confirmatory, hence information from clinical, radiological, physiological, and immunological evaluations may be used together for a confirmative diagnosis of hypersensitivity pneumonitis. The treatment of choice is avoidance of antigen but systemic corticosteroids may be effective in suppressing the inflammatory response. The prognosis depends on early diagnosis and effective antigen avoidance.

Immune response to n-terminal and c-terminal deletion mutants of Aspergillus fumigatus major allergen ASP F 3.

The ubiquitous fungus Aspergillus fumigatus causes allergic rhinitis, asthma, sinusitis and allergic bronchopulmonary aspergillosis. A number of major allergens from A. fumigatus are purified, but their structure-function role in the pathogenesis of disease is not known. Such information is essential for devising alternative therapy of fungal allergic diseases. In the present study, N-terminal and C-terminal deletion mutants ofAsp f 3 were constructed and their immunopathological responses studied in a mice model of allergy. Three mutants viz,Asp f 3 (aa 33-168), (aa 1-142), and (aa 23-142) were made by deleting certain amino acids from epitopic regions of full lengthAsp f 3, a major allergen of A. furnigatus. TheAsp f 3 and three mutated proteins were expressed in pET vector. The C-terminal deletion mutantAsp f 3 (aa 1-142) induced elevated IFN-γ but low levels of IL-4 by spleen cells. This mutant also showed significant downregulation of peripheral blood eosinophils and lung inflammation in immunized mice. The N-terminal deletion mutantAsp f 3 (aa 33-168) also exhibited an immuno-suppressive effect in terms of IgE production and induction of Th2 cytokine. The results indicate thatrAsp f 3 and its deletion mutants induced distinct immune-inflammatory responses in mice on challenge with these proteins. The non-IgE binding deletion mutants ofAsp f 3 (aa 1-142 and aa 33-168) could deviate Th2 immune response with a concomitant reduction in airway inflammation and infiltration of inflammatory cells.

Microarrayed allergen molecules: diagnostic gatekeepers for allergy treatment.

Type I allergy is an immunoglobulin E (IgE)-mediated hypersensitivity disease affecting more than 25% of the population. Currently, diagnosis of allergy is performed by provocation testing and IgE serology using allergen extracts. This process defines allergen-containing sources but cannot identify the disease-eliciting allergenic molecules. We have applied microarray technology to develop a miniaturized allergy test containing 94 purified allergen molecules that represent the most common allergen sources. The allergen microarray allows the determination and monitoring of allergic patients' IgE reactivity profiles to large numbers of disease-causing allergens by using single measurements and minute amounts of serum. This method may change established practice in allergy diagnosis, prevention, and therapy. In addition, microarrayed antigens may be applied to the diagnosis of autoimmune and infectious diseases.

Severe allergic reactions to guinea pig.

BACKGROUND: Allergic sensitization and reactions to guinea pig (Cavia porcellus) have been well documented in laboratory animal handlers, primarily manifesting as rhinitis, conjunctivitis, and asthma. Severe allergic reactions, however, are rare. METHODS: We report two patients with severe allergic reactions following non-occupational exposure to guinea pigs. The first patient, an 11-year-old female, developed ocular, nasal, skin and laryngeal edema symptoms immediately after handling a guinea pig. The second patient, a 24-year-old female, developed symptoms of isolated laryngeal edema after cleaning a guinea pig cage. Percutaneous skin testing, RAST, ELISA and ELISA inhibition testing with guinea pig extract were performed. RESULTS: Both patients had IgE-mediated allergy to guinea pig confirmed by ELISA and either RAST or skin testing. ELISA inhibition studies confirmed the specificity of the IgE reactivity to guinea pig. CONCLUSION: Severe IgE-mediated reactions can occur following non-occupational guinea pig exposure. Physicians should be aware of this possibility.

Specific IgE response to purified and recombinant allergens in latex allergy.

BACKGROUND: In recent years, allergy to natural rubber latex has emerged as a major allergy among certain occupational groups and patients with underlying diseases. The sensitization and development of latex allergy has been attributed to exposure to products containing residual latex proteins. Although improved manufacturing procedures resulted in a considerable reduction of new cases, the potential risk for some patient groups is still great. In addition the prevalent cross-reactivity of latex proteins with other food allergens poses a major concern. A number of purified allergens and a few commercial kits are currently available, but no concerted effort was undertaken to evaluate them. METHODS: We studied 11 purified latex allergens, Hev b 1 to Hev b 10, and Hev b 13 along with several crude allergen extracts and two commercial ImmunoCAP assays to evaluate specific IgE antibody in the sera from latex allergic patients and controls. Health care workers and spina bifida patients with clinical symptoms of latex allergy, spina bifida patients without latex allergy, and non-atopic health care workers have been studied. RESULTS: The results suggest that Hev b 2, 5, 6, and 13 together identified over 80 percent health care workers with latex allergy, while Hev b 6 along with Hev b 1 or 3 detected specific IgE antibody in all sera studied from patients with spina bifida and latex allergy. The ImmunoCAP results using both Hev b 5 amplified and non-amplified closely agreed with the clinical diagnosis of latex allergy in health care workers and in spina bifida. CONCLUSION: Although the purified allergens and crude extracts reacted diversely with IgE from different patient groups, the results indicated that use of certain combinations of purified recombinant antigens will be useful in commercial kits or in in-house assays for detecting specific IgE antibody in the sera. The results suggest that a combination of Hev b 2, 3, 5, 6, and 13 together detected specific IgE in 80% of the sera from latex allergic patients. Both ImmunoCAPs correctly identified over 95% of latex allergic patients, however, showed reactivity with a few normal control subjects.

Allergic bronchopulmonary aspergillosis in cystic fibrosis--state of the art: Cystic Fibrosis Foundation Consensus Conference.

Because of the difficulties of recognizing allergic bronchopulmonary aspergillosis (ABPA) in the context of cystic fibrosis (because of overlapping clinical, radiographic, microbiologic, and immunologic features), advances in our understanding of the pathogenesis of allergic aspergillosis, new possibilities in therapy, and the need for agreed-upon definitions, an international consensus conference was convened. Areas addressed included fungal biology, immunopathogenesis, insights from animal models, diagnostic criteria, epidemiology, the use of new immunologic and genetic techniques in diagnosis, imaging modalities, pharmacology, and treatment approaches. Evidence from the existing literature was graded, and the consensus views were synthesized into this document and recirculated for affirmation. Virulence factors in Aspergillus that could aggravate these diseases, and particularly immunogenetic factors that could predispose persons to ABPA, were identified. New information has come from transgenic animals and recombinant fungal and host molecules. Diagnostic criteria that could provide a framework for monitoring were adopted, and helpful imaging features were identified. New possibilities in therapy produced plans for managing diverse clinical presentations.

Vaccination of mice against invasive aspergillosis with recombinant Aspergillus proteins and CpG oligodeoxynucleotides as adjuvants.

In a murine model of invasive pulmonary aspergillosis, dendritic cells (DCs) pulsed with Aspergillus antigens induced the activation of CD4(+) Th1 cells capable of conferring resistance to the infection. Here we show that the combined, local delivery of unmethylated CpG oligodeoxynucleotides (ODNs) and the Asp f 16 Aspergillus allergen resulted in the functional maturation and activation of airway DCs capable of inducing Th1 priming and resistance to the fungus. Therefore, ODNs act as a potent adjuvant for the vaccine-induced protection against the fungus by promoting dominant Th1 response to Aspergillus antigens and allergens.

Animal models of allergic bronchopulmonary aspergillosis.

Experimental animal models of Allergic Bronchopulmonary Aspergillosis (ABPA) serve several purposes. Both common and distinct pathological features occurring in natural and experimental diseases are of great interest as they serve to identify the key elements in the pathogenesis. Experimentally induced diseases can be modeled to understand the various parameters such as antigen and route of exposure, genetic background and the role of response modifiers in the disease process. Furthermore, animals with targeted gene-deletion or with insertion of transgenes have been studied to define the roles of specific cells, receptors and mediators in the pathogenesis. The resulting conclusions have been used to formulate hypothesis, which have to be tested for their application to human disease.

Molecular biology of Aspergillus allergens.

Allergic Bronchopulmonary Aspergillosis (ABPA) is a severe respiratory disease caused by the fungus Aspergillus fumigatus (Af). It occurs as secondary complication mostly in patients with atopy and cystic fibrosis. The standardized and well-characterized allergens are essential for the immunodiagnosis of ABPA as well as for understanding the pathophysiology of the disease. Molecular cloning was resorted to obtain purified Af allergens for such studies. Currently, twenty-two recombinant Af allergens have been identified and characterized and several of these can be used as standardized allergens in in vitro and in vivo diagnosis of ABPA. The knowledge of primary, secondary, and tertiary structures of these allergens may facilitate the identification of immunodominant T and B cell epitopes and may be used to unravel the structure function relationship of these allergens. Such findings may open up novel avenues in the immune modulatory therapy and other effective intervention of the disease.

Aspergillus antigens associated with allergic bronchopulmonary aspergillosis.

Among the molds causing allergy, Aspergillus fumigatus (Af) constituted a major species present both indoors and outdoors. The antigens both secreted and bound to hyphae and conidia have been isolated for immunodiagnosis of allergic aspergillosis. The crude extracts have been used to demonstrate IgE antibody in vitro and for skin testing. The crude extracts contain many antigenic and non-antigenic components and toxins and have demonstrated inconsistent reactivities. In addition, similarity with other allergens also may complicate specific diagnosis. A number of methods including conventional purification and fractionation methods have been used to obtain relevant antigens. In recent years, monoclonal antibody dependent affinity purification and molecular biology methods have obtained considerable progress in the allergen purification and in developing specific and reliable immunoassays. However, international standards are still lacking and hence, comparison of results among laboratories are not possible yet.

Immunodiagnosis of ABPA.

For the diagnosis of allergic aspergillosis demonstration of specific immune responses to allergens has been accepted as a significant paradigm. Elevated levels of total IgE and Aspergillus fumigatus specific IgE and IgG antibodies are important criteria for diagnosis of ABPA. Although reference antigens or standardized methods are not available, there are a number of relevant recombinant antigens, which have been isolated in recent years. Several techniques have been employed in the demonstration of specific antibodies against antigens of Aspergillus fumigatus in the sera of patients. Of these methods, the widely followed ones are ELISA, radioimmunoassay, Western blot, and agar gel double diffusion. Recently, semi-automated methods have been developed using recombinant allergens to detect circulating antibody against Aspergillus. However, these methods have not been evaluated widely. Here we review the immunodiagnostic methods currently in use for allergic bronchopulmonary aspergillosis.

IgE binding synthetic peptides of Alt a 1, a major allergen of Alternaria alternata.

Alternaria alternata protein, Alt a 1 is a major allergen associated with allergy in atopic patients. Although the molecule binds strongly to IgE antibody from patients, the epitopes involved have not been identified or defined. In the present study, we synthesized overlapping peptides spanning the whole sequence and evaluated their IgE binding with sera from patients with Alternaria-induced allergy. The results identified four IgE binding linear regions. Two of these regions K41-P50 and Y54-K63 showed consistent reactivity with all four patients studied. The specific epitopes involved in the immune response may be of value in the immunodiagnosis and probably also in specific immunotherapy.

Detection of latex allergens by immunoelectron microscopy in ambient air (PM10) in Oslo, Norway (1997-2003).

The authors collected ambient air along two highways in Oslo to investigate the annual variations in particulate matter (PM10) and the presence of latex as an outdoor allergen. PMI, was monitored for a period of five years, during which time the use of studded winter tires was reduced. The presence of latex and of common aeroallergens was examined directly on the collection filters with immunoelectron microscopy visualized in a scanning electron microscope. The annual variation in PM10 was similar over the five years of sampling, with increased mass concentrations in winter. Statistical analysis indicated no major effect from the change to nonstudded tires. The most important factors influencing the PM10 concentration were meteorological parameters like wind and rain. Immnunolabeling of the filters showed latex as an outdoor allergen that adhered to carbon aggregates from vehicle emission. The results also indicated cross-reactive epitopes among the common allergens investigated, which for sensitized subjects may add to the risk of developing latex allergy.

Immunomodulatory effects of curcumin in allergy.

Recent years have witnessed a global increase in allergy and asthma, particularly in developed countries. Attempts to develop effective control measures for allergy and asthma resulted in the exploration of alternate medicines including herbal remedies traditionally used in old world countries. Turmeric is known for its multiple health restoring properties, and has been used in treating several diseases including several respiratory disorders. Turmeric is a common spice used in the culinary preparations in South and East Asian countries. The active component of turmeric is curcumin, a polyphenolic phytochemical, with anti-inflammatory, antiamyloid, antiseptic, antitumor, and antioxidative properties. Curcumin was reported to have antiallergic properties with inhibitory effect on histamine release from mast cells. The effectiveness of curcumin in allergy and asthma has been further investigated using a murine model of allergy. The results indicate a marked inhibition of allergic response in animals treated with curcumin suggesting a major role for curcumin in reducing the allergic response. The present review focuses on the results of research aimed to understand the immunomodulation induced by curcumin and its associated roles in the amelioration of allergy. These findings needed further evaluation, extrapolation, and confirmation before using curcumin for controlling allergy and asthma in humans.

Pulmonary arterial remodeling induced by a Th2 immune response.

Pulmonary arterial remodeling characterized by increased vascular smooth muscle density is a common lesion seen in pulmonary arterial hypertension (PAH), a deadly condition. Clinical correlation studies have suggested an immune pathogenesis of pulmonary arterial remodeling, but experimental proof has been lacking. We show that immunization and prolonged intermittent challenge via the airways with either of two different soluble antigens induced severe muscularization in small- to medium-sized pulmonary arteries. Depletion of CD4(+) T cells, antigen-specific T helper type 2 (Th2) response, or the pathogenic Th2 cytokine interleukin 13 significantly ameliorated pulmonary arterial muscularization. The severity of pulmonary arterial muscularization was associated with increased numbers of epithelial cells and macrophages that expressed a smooth muscle cell mitogen, resistin-like molecule alpha, but surprisingly, there was no correlation with pulmonary hypertension. Our data are the first to provide experimental proof that the adaptive immune response to a soluble antigen is sufficient to cause severe pulmonary arterial muscularization, and support the clinical observations in pediatric patients and in companion animals that muscularization represents one of several injurious events to the pulmonary artery that may collectively contribute to PAH.