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L K Arruda

Publications and source records attributed to L K Arruda.

At least 37 records · Page 2Linked to original sources
PubMed1997

Evaluation of German cockroach (Orthoptera:Blattellidae) allergen and seasonal variation in low-income housing.

Six apartments in a low-income housing project were evaluated for German cockroach. Blattella germanica (L.), infestation and concentration of an allergen derived from these cockroaches (Bla g II). Kitchen and living room samples were collected monthly for 1 yr. In addition, airborne sampling was carried out in 5 kitchens. The kitchen had the highest allergen concentration in 65% of visits and the highest number of cockroaches trapped in 69% of visits. In the kitchen, the highest cockroach levels were seen in June, whereas the values for Bla g II peaked in August. In keeping with this, the closest correlation was between Bla g II (microgram/g dust) and the number of cockroaches found 2 mo earlier. Airborne samples were assayed for 2 separate allergens. Bla g II and Bla g I. No allergen was detectable in the absence of disturbance. By contrast, during disturbance with a vacuum cleaner both Bla g II and Bla g I were detectable in the air of each apartment. Results suggest that immunochemical assay of a major allergen in dust samples from the kitchen floor may be used to monitor exposure to German cockroaches, also that cockroach levels may be used as an indicator or predictor of allergen in dust.

Allergens↗
PubMed1997

Recombinant mite allergens. New technologies for the management of patients with asthma.

The introduction of molecular cloning techniques has led to advances in allergen identification and sequencing, production of recombinant allergens, identification of B-cell and T-cell epitopes, and tertiary structural analysis of allergen molecules. Over 10 groups of mite allergens have been cloned from Dermatophagoides spp., as well as several homologous allergens from Euroglyphus maynei, Lepidoglyphus destructor, and Blomia tropicalis. The availability of these allergens has made it feasible to consider their use for both diagnostic and therapeutic purposes. Several recombinant Dermatophagoides allergens show comparable reactivity on skin testing and in serologic assays to natural allergens, and cocktails of the recombinant proteins could be used as diagnostic reagents. New technologies have been developed for detection of allergen-specific IgE and for environmental allergen detection using rapid diagnostic tests. Novel approaches to immunotherapy are also being investigated, including T-cell-peptide based vaccines, allergen variants which lack IgE reactivity, and naked DNA vaccines. The application of allergen biotechnology should lead to improvements in the management of mite-allergic patients with asthma and represents a logical step toward reducing asthma mortality and morbidity.

Allergens↗
PubMed1997

Sensitization to Blomia tropicalis in patients with asthma and identification of allergen Blo t 5.

In tropical and subtropical regions of the world, allergens produced by Blomia tropicalis are an important cause of IgE-mediated sensitization among patients with asthma. We compared the relative importance of sensitization to the two mite species among asthma patients from Florida, Puerto Rico, and Brazil (n = 83), who were concurrently exposed to B. tropicalis and D. pteronyssinus, with patients from the United States and from the United Kingdom (n = 56) exposed to D. pteronyssinus. In addition, molecular cloning techniques were used to clone and express a major B. tropicalis allergen. There were significant differences between IgE antibody responses to B. tropicalis and D. pteronyssinus that were related to exposure: only 22% of patients exposed to both species had a high ratio (> 10) of IgE D. pteronyssinus:B. tropicalis, whereas 68% of patients exposed only to D. pteronyssinus had a ratio of > 10 (p < 0.001). A major 14-kD allergen (Blo t 5), cloned from a B. tropicalis cDNA library, showed 43% sequence homology to D. pteronyssinus Der p 5. Recombinant Blo t 5 produced in E. coli reacted with 45 to 69% of sera from B. tropicalis-allergic asthmatics and induced positive immediate skin tests at 10(-3) to 1 microg/ml. In vivo and in vitro comparisons of IgE responses to B. tropicalis, D. pteronyssinus, rBlo t 5, and rDer p 5, showed that B. tropicalis has unique allergens that cause specific IgE responses. The results suggest that B. tropicalis is an independent cause of sensitization and that use of recombinant Blo t 5 should lead to a better understanding of the role of B. tropicalis in causing asthma in tropical environments.

Adolescent↗
PubMed1995

Cloning of cockroach allergen, Bla g 4, identifies ligand binding proteins (or calycins) as a cause of IgE antibody responses.

An allergen cloned from a Blattella germanica (German cockroach) cDNA library, encoded a 182-amino acid protein of 20,904 Da. This protein, designated B. germanica allergen 4 (Bla g 4), was expressed as a glutathione S-transferase fusion protein in Escherichia coli and purified by affinity chromatography and high-performance liquid chromatography. The prevalence of serum IgE antibody to recombinant Bla g 4 in 73 cockroach allergic patients with asthma ranged from 40% (antigen binding radioimmunoassay) to 60% (plaque immunoassay). Cockroach allergic patients gave positive intradermal skin tests to recombinant Bla g 4 at concentrations of 10(-3)-10(-5) micrograms/ml, whereas non-allergic controls, or cockroach allergic patients with no detectable serum IgE antibody to Bla g 4, gave negative skin tests to 1 microgram/ml. Polymerase chain reaction and Southern analysis identified a 523-base pair DNA encoding Bla g 4 in both B. germanica and Periplaneta americana (American cockroach). However, Northern analysis showed that mRNA encoding Bla g 4 was transcribed in B. germanica but not in P. americana, suggesting that allergen expression was species specific. Sequence similarity searches showed that Bla g 4 was a ligand binding protein or calycin and unexpectedly revealed that this family contained several important allergens: beta-lactoglobulin, from cow milk, and rat and mouse urinary proteins. Although the overall sequence homology between these proteins was low (approximately 20%), macromolecular modeling techniques were used to generate two models of the tertiary structure of Bla g 4, based on comparisons with the x-ray crystal coordinates of bilin binding protein and rodent urinary proteins. The results show that members of the calycin protein family can cause IgE antibody responses by inhalation or ingestion and are associated with asthma and food hypersensitivity.

Allergens↗
PubMed1995

Molecular cloning of a major cockroach (Blattella germanica) allergen, Bla g 2. Sequence homology to the aspartic proteases.

Inhalation of allergens produced by the German cockroach (Blattella germanica) elicits IgE antibody formation and the development of asthma in genetically predisposed individuals. We compared the allergenic importance of two cockroach (CR) allergens, Bla g 1 and Bla g 2, and determined the complete amino acid sequence of the major 36-kDa allergen, Bla g2. A survey of 106 sera from CR allergic patients showed the prevalence of IgE antibodies to Bla g 1 and Bla g 2 to be 30.2% and 57.6%, respectively. Immediate skin tests on 7 selected patients gave positive reactions using 10(-3) micrograms/ml either allergen, whereas controls showed no response to 10 micrograms/ml. Natural Bla g 2 was purified and the sequence of the NH2 terminus and tryptic peptides, comprising 36% of the molecule, was determined. The cDNA for Bla g 2 was cloned from a B. germanica expression library and encoded a 24-amino acid signal peptide and a 328-amino acid mature protein, which showed the highest degree of identity to mosquito (Aedes aegypti) lysosomal aspartic protease (30.8%), with similar identity to pepsin, cathepsins D and E, renin, and chymosin. Bla g 2 mRNA and protein were detected in B. germanica, but not in Periplaneta americana, the other principal domiciliary CR species in the U.S. High concentrations of Bla g 2 were found in CR digestive organs (esophagus, gut, and proventriculus). The results show that Bla g 2 is a major species-specific allergen of B. germanica and suggest that the allergen functions as a digestive enzyme in the cockroach.

Allergens↗
PubMed1994

Development of immune responses to Aspergillus at an early age in children with cystic fibrosis.

Although the ability of Aspergillus organisms to colonize the respiratory tract in patients with cystic fibrosis (CF) is well recognized, the contribution of Aspergillus to the disease process is poorly understood. Using sera from 147 CF patients (age 5 to 43 yr), we measured IgE antibody (ab) to Aspergillus fumigatus and five common inhalant allergens with a radioallergosorbent test (RAST). Total IgE levels and IgG ab to radio-labeled Asp f I, an allergen purified from A. fumigatus and a potent inhibitor of protein synthesis, were also measured. Thirty (20%) of the patients had IgE ab to A. fumigatus, and 22 (15%) of these patients had developed total IgE levels > or = 400 IU/ml, raising the consideration of a diagnosis of allergic bronchopulmonary aspergillosis (ABPA). Five of the 22 patients developed these IgE responses by age 5 yr and 14 by age 10 yr. The proportion of patients with IgE ab to one or more of the other allergens tested was not significantly different from that of control subjects without respiratory symptoms. A striking proportion (84%) of CF sera contained IgG ab to Asp f I, compared with 6% of sera from control patients and 20% of sera from allergic children with asthma (n = 25), only one of whom had IgE ab to A. fumigatus. In an examination of additional sera from young CF patients, IgG anti-Asp f I ab was detected in 41% of these sera from patients 5 yr of age or older, increasing to 98% of 89 sera from patients older than age 10.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1993

A transformant of Aspergillus fumigatus deficient in the antigenic cytotoxin ASPFI.

The aspfI gene encoding a ribonucleotoxin, a putative virulence factor of Aspergillus fumigatus, was inactivated by gene disruption. Gene replacement through homologous recombination by the disrupted allele tagged by the hygromycin B resistance marker was performed by transformation of a pathogenic strain. One transformant with the disrupted aspfI gene failed to produce the ASPFI protein and was shown to be pathogenic for mice. We concluded that this ribotoxin is not a main factor in the colonization of the lung tissues by A. fumigatus.

Allergens↗
PubMed1993

Environmental exposure to Aspergillus fumigatus allergen (Asp f I).

Asp f I is a major allergen produced by the mycelia of Aspergillus fumigatus. It is not present in spores and can be used as a specific marker for the detection of germination of this fungus. We investigated the domestic and outdoor concentration of Asp f I in Poole, U.K. and Charlottesville, VA, U.S.A. Asp f I was undetectable in 95% (281/296) of house dust extracts and present at low levels (< 0.17 micrograms/g of sieved dust, mean 0.038 micrograms/g) in the remainder. In contrast, Asp fI could be detected in 65% (15/23) of cultures of house dust, suggesting the presence of viable, but ungerminated, A. fumigatus in the majority of homes. Asp f I was detectable in 80% (28/35) of extracts of leaves and compost, but present in these outdoor samples at low levels (< 0.11 micrograms/g, mean 0.27 micrograms/g). Air sampling for Asp f I was undertaken before and after vigorous disturbances at indoor (n = 5) and outdoor (n = 6) sites. Airborne Asp f I was not detected in domestic samples or in undisturbed outdoor samples. Following disturbance it could be measured in outdoor samples (range 7.6-29 ng/m3). The results suggest that while exposure to A. fumigatus is common, exposure to Asp f I and germinating A. fumigatus is uncommon. It is probable that those individuals who develop antibody responses to Asp f I have been exposed to A. fumigatus which has germinated in their respiratory tract.

Air↗
PubMed1993

IgG and IgE antibody responses to dust mite allergens among children with asthma in Brazil.

We compared exposure and levels of IgE and IgG antibodies to inhalant allergens in 20 children with asthma and positive skin tests to Dermatophagoides pteronyssinus, living in São Paulo, to 20 control children without history of asthma or allergy and negative skin tests to D. pteronyssinus, D. farinae, and Blomia tropicalis. In 16/20 and 17/20 houses of asthmatic and control children, respectively, at least one sample was obtained which contained > 10 micrograms Der p I/g of dust. Serum IgE antibodies to D. pteronyssinus and Blomia tropicalis were > 200 RAST U/mL in 19/20 and in 16/20 asthmatic children, respectively. In the control group, IgE antibodies to either mite species were < 40 RAST U/mL in most cases. IgG anti-Der p I and anti-Der f I antibodies were detected in 17/20 asthmatics, as opposed to 3/20 controls. Neither cat nor cockroach allergens caused significant sensitization among asthmatic or control children. Although exposure to high levels of mite allergens was common in São Paulo, significant IgE and IgG antibody responses were detected only in children with asthma.

Administration, Inhalation↗
PubMed1992

Selective expression of a major allergen and cytotoxin, Asp f I, in Aspergillus fumigatus. Implications for the immunopathogenesis of Aspergillus-related diseases.

Asp f I is a major 18-kDa Aspergillus fumigatus allergen and a member of the mitogillin family of cytotoxins. The nucleotide sequence of the Asp f I gene was determined by sequencing polymerase chain reaction products amplified from A. fumigatus spore DNA. The entire 678-bp DNA includes an 81-bp leader sequence, preceding the N-terminal alanine codon, a 52-bp intron, and a 444-bp open reading frame, encoding a 149-amino acid protein (M(r) 16,899), which is 99% homologous to mitogillin from Aspergillus restrictus. A mAb-based ELISA was used to compare Asp f I levels in spores, mycelia, and culture filtrate, and to determine the kinetics of allergen production. Disrupted hyphae or spore extracts had a 1000-fold lower level of Asp f I than culture filtrate, suggesting that germination of spores and growth of the fungus are essential for allergen production. Asp f I levels in A. fumigatus and A. restrictus peaked at day 3 (0.87 to 12.1 micrograms/ml), however, the allergen was not detected in Aspergillus flavus, Aspergillus niger, Aspergillus terreus, and Aspergillus nidulans cultures (< 1.5 ng/ml) on either days 3 or 8. Northern analysis confirmed that Asp f I mRNA was detected only in A. fumigatus and A. restrictus, but not in the other four Aspergillus spp. Asp f I-specific DNA was generated after polymerase chain reaction amplification of genomic mycelial DNA obtained from A. fumigatus and A. restrictus, but not from the other Aspergillus spp. The results show that Asp f I is selectively expressed in A. fumigatus, and suggest that this cytotoxin could be a specific virulence factor for A. fumigatus.

Allergens↗
PubMed1992

A review of recent immunochemical studies of Blomia tropicalis and Euroglyphus maynei allergens.

Exposure to mites other than Dermatophagoides spp., particularly Blomia tropicalis and Euroglyphus maynei, has been increasingly recognized as a cause of asthma. Positive skin tests and serum IgE antibodies to B. tropicalis have been reported in asthmatic patients from several areas of the world, including São Paulo (Brazil), Hong Kong and Tampa (Florida, USA). Analysis of B. tropicalis extracts showed undetectable levels of the major Group I and Group II Dermatophagoides spp. allergens. Immunoabsorption experiments showed that most of the IgE antibodies to B. tropicalis (approximately 70%) reacted with species-specific allergens. Murine monoclonal antibodies to B. tropicalis could present antigens that were recognized by human IgG antibodies. Sensitization to E. maynei has been reported in Europe, North and South America and Australia. Analysis of four different E. maynei extracts by ELISA and RIA showed that E. maynei produces an allergen that is antigenically related to Dermatophagoides Group I allergens. The amino acid sequence of this allergen (Eur m I) has recently been reported. Further identification and purification of B. tropicalis and E. maynei allergens is required to develop specific assays for measuring these allergens in dust samples. This will make it possible to investigate the relationship between exposure to B. tropicalis or E. maynei and the development of sensitization and allergic disease.

Allergens↗
PubMed1992

Aspergillus fumigatus: identification of 16, 18, and 45 kd antigens recognized by human IgG and IgE antibodies and murine monoclonal antibodies.

The immunochemical properties of antigens produced by Aspergillus fumigatus were investigated with biochemical purification techniques in conjunction with the production of murine monoclonal antibodies (MAbs) and binding studies with human IgG and IgE antibodies. A. fumigatus antigens were partially purified by gel filtration and hydrophobic interaction chromatography on phenyl-Sepharose. Two fractions that eluted with either 2 mol/L or 0.15 mol/L of NaCl demonstrated strong binding to human IgG and IgE antibodies. Immunoprecipitation analysis with IgG antibodies from six patients with different Aspergillus-related diseases demonstrated that the 2M and 0.15M fractions contained major antigens of molecular weight 18 kd (Asp f I) and 45 kd, respectively. The 125I-labeled 2M fraction was used to compare IgG antibodies to A. fumigatus in sera from 25 patients with Aspergillus-related diseases. IgG antibodies were significantly higher in patients with allergic bronchopulmonary aspergillosis (geometric mean, 437 U/ml) than in patients with asthma (geometric mean, 14 U/ml; p less than 0.001), but undetectable (less than 5 U/ml) in 43/48 control subjects. A good correlation was found between levels of IgG antibodies to the 125I-labeled 0.15M fraction and the 125I-labeled 2M fraction in sera from 106 patients with cystic fibrosis (r = 0.77; p less than 0.001). Five murine IgG MAbs and two IgM MAbs were raised against the 2M fraction, and immunoprecipitation with the IgG MAb demonstrated two distinct antigens within the 2M fraction, Asp f I, and a 16 kd antigen. The results of a solid-phase RIA with IgG MAb 4A6 demonstrated that approximately 85% of A. fumigatus-allergic patients with allergic bonchopulmonary aspergillosis had IgE antibodies to Asp f I. The three protein antigens defined in these studies are useful probes for investigating the immunopathogenesis of diseases associated with colonization by A. fumigatus.

Animals↗
PubMed1992

Aspergillus epidural abscess and cord compression in a patient with aspergilloma and empyema. Survival and response to high dose systemic amphotericin therapy.

A 57-yr-old man with a chronic lung cavity presumed to be related to ankylosing spondylitis and/or old cavitary tuberculosis presented with hemoptysis and rapidly developed lower extremity paresis and hypoesthesia. On chest radiograph he had a left upper lobe lesion suggestive of aspergilloma combined with a large left empyema with bronchopleural fistula. Serologic analysis demonstrated precipitins and very high titer IgG antibodies to Aspergillus fumigatus antigens. Decompressive laminectomy from T1 to T5 was performed, with drainage of A. fumigatus culture-positive material from an epidural abscess compressing the spinal cord. Chest drainage was required for control of the empyema. With a total course of 3 g of intravenously administered amphotericin B, rehabilitative therapy, and chronic empyema drainage, he is now at home and ambulatory with assistance. He is also being followed by regular serum assays of IgG antibodies to Aspergillus proteins. We report the case of an apparent long-term survivor of a formerly lethal and/or nonreversible paraplegic condition. The critical factors compared with previous cases with a poor outcome would appear to be prompt neurosurgical intervention, restoration of a normal number of T-cells, effective long-term chest drainage, and high dose amphotericin treatment.

Abscess↗
PubMed1991

Trichophyton tonsurans allergen. I. Characterization of a protein that causes immediate but not delayed hypersensitivity.

Fungal infections of skin or nails are extremely common and often caused by dermatophyte fungi of the genus Trichophyton. These fungi are unusual in that they can give rise to delayed hypersensitivity (DH) or immediate hypersensitivity (IH) responses. Recently, IH to Trichophyton tonsurans has been demonstrated in patients by skin tests, serum IgE antibody test (RAST), and positive nasal and bronchial challenges. To further investigate the immunology of Trichophyton, a 30-kDa T. tonsurans allergen was isolated by gel filtration and hydrophobic interaction chromatography. This protein, Tri t I, gave a single band on SDS-PAGE, and the 30 amino-terminal amino acids have been determined. Among patients with positive IH skin tests, 34 of 48 (71%) had IgG antibody and 26 of 48 (54%) had IgE antibody to Tri t I. Among those who had positive responses to both skin tests and RAST, 22 of 30 (73%) had IgE antibodies to Tri t I; thus, this protein represents a major allergen. Twelve clones of murine IgG mAb antibodies were produced. Two clones, 2F2-F7 and 6B11-C2, were found to define separate epitopes on Tri t I and were used to develop an immunometric assay for the quantitation of Tri t I. Twenty-three of 38 volunteers with a history of athlete's foot were found to have either IH and/or DH to Trichophyton mix and underwent further testing with purified Tri t I. Of the nine found to have IH to the mix, eight were sensitive to Tri t I. Seven of these eight had IgG and IgE antibodies to Tri t I, by Ag-binding RIA, and all were RAST positive to the unpurified extract. An additional 14 had either DH alone (n = 7) or a wheal and flare response followed by DH at 48 h (n = 7). Of these 14 who had DH responses to Trichophyton mix, only one showed DH to an equivalent quantity of purified Tri t I; among this group, none showed IH or serum IgE antibodies and only one had detectable IgG antibody to Tri t I. The results suggest that the majority of subjects with DH to Trichophyton are responding to a protein other than Tri t I and that the wheal that precedes DH reactions is some patients is not associated with IgE antibodies.

Allergens↗