PubMed Health⌕ Search

Biomedical subjects

M D Chapman

Publications and source records attributed to M D Chapman.

At least 19 recordsLinked to original sources
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

Benzyl benzoate moist powder: investigation of acaricidal [correction of acarical] activity in cultures and reduction of dust mite allergens in carpets.

Despite advances in the understanding of dust mites, it remains difficult to control exposure to mite allergens, and it is particularly difficult to reduce mites in fitted carpets or sofas. Several chemicals have been demonstrated to kill mites or denature mite allergens, and some of these chemicals have been investigated in carpets. Benzyl benzoate (BB), which has been widely used to kill scabies mites and is known to kill mites of the genus Dermatophagoides, has been used as a method of treating carpets. The present article describes experiments in the laboratory and in houses in testing two preparations of BB, a moist powder and a foam. The moist powder is composed of two ingredients, a wetted "inert" cellulose, which is designed to act as a cleaning agent, and the active BB adsorbed onto silicates. The active powder kills 90% of mites in culture within 12 hours and 100% in 24 hours, whereas the cellulose is not acaricidal. The moist-powder preparation was highly effective at killing D. farinae and D. pteronyssinus mites in the laboratory. In carpets the moist powder, applied for 12 hours with repeated brushing, was demonstrated to reduce the concentrations of group I and group II dust mite allergens in dust recovered at 1 month. This decrease in concentration could, in part, be explained by a persistent increased recovery of dust caused by residual white powder. However, when the recovery of group II allergens was calculated as the total allergen recovered, the decrease was highly significant at 2 weeks and 4 weeks after treatment (p less than 0.001). Application of the powder to carpets for 4 hours or of the foam to sofas was less effective. After 2 months the effect on mite antigen in carpets was still present, but some increase was apparent, suggesting that repeat application after 2 or 3 months would be necessary to control mite-allergen levels.

Allergens↗
PubMed1992

Risk factors for asthma in inner city children.

Inner city children have the highest prevalence and the highest mortality rates for asthma in the United States. The purpose of this study was to evaluate sensitization and exposure to common indoor allergens among children aged 3 years to 15 years seen for treatment of asthma at Grady Memorial Hospital, Atlanta, Ga. Eighty children in this study were enrolled in the emergency department and 64 in hospital clinics. Dust from 57 homes, assayed for three indoor allergens (dust mite, cat, and cockroach), revealed similar exposure for asthma and control groups. Sixty-nine percent of the children with asthma had IgE antibodies to dust mite, cockroach, or cat; only 27% of the control subjects were similarly sensitized (p < 0.001). Of 35 children with asthma 21 had both sensitization and significant exposure to the relevant allergen; this was true for only 3 of 22 control subjects (odds ratio, 9.5; p < 0.001). Neither sensitization nor exposure to cat allergen was common in this population. The results show that black children in inner city Atlanta are exposed to high levels of mite and cockroach allergens and that a high proportion of the children with asthma are sensitized to these allergens; the combination of sensitization and exposure is a major risk factor for asthma in this population.

Adolescent↗
PubMed1992

Cat allergy.

Explore the source record for details and available documents.

Allergens↗
PubMed1991

Amino acid sequence of Fel dI, the major allergen of the domestic cat: protein sequence analysis and cDNA cloning.

The complete primary structure of Fel dI (International Union of Immunological Societies nomenclature), the major allergen produced by the domestic cat, Felis domesticus, was determined by protein sequence analysis and cDNA cloning. Protein sequencing of Fel dI from an immunoaffinity-purified extract of house dust revealed that the allergen is composed of two polypeptide chains. Degenerate oligonucleotides derived from the protein sequence were used in polymerase chain reaction amplification of cat salivary gland cDNA to demonstrate that the two chains are encoded by different genes. Chain 1 of Fel dI shares amino acid homology with rabbit uteroglobin, while chain 2 is a glycoprotein with N-linked oligosaccharides.

Allergens↗
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↗
PubMed1991

Environmental exposure to cockroach allergens: analysis with monoclonal antibody-based enzyme immunoassays.

Quantitative two-site monoclonal antibody (MAb)-based enzyme-linked immunoassays for two cockroach (CR) allergens, Bla g I and Bla g II, have been developed and used to measure allergen levels in house-dust samples. Dust collected from the CR-infested homes of two patients with asthma from Charlottesville, Va., demonstrated wide variation in the levels of Bla g I, depending on the location of dust collection. Dust from kitchen floors and cabinets contained 50-fold more allergen (mean, 10,755 U/gm of dust) than dust from bedrooms and upholstered furniture (mean, 204 U/gm). One hundred forty-five dust samples were collected from the bedrooms and living rooms of 22 children with asthma and 16 control subjects without asthma living in Atlanta, Ga. Twenty-seven of the 38 homes (17/22 children with asthma; 10/16 control subjects) had detectable Bla g I (4 to 1340 U/gm of dust). Bla g II levels were assayed in 40 kitchen, bedroom, and living room samples from homes in Wilmington, Del. Highest levels of Bla g II were detected in kitchen-floor dust (300 U/gm of dust). Additionally, approximately 20% of homes with no visual evidence of CR infestation had significant levels of Bla g II in at least one dust sample (greater than 4 U/gm of dust). Our results demonstrate that CR may be an occult allergen in homes. The kitchen appears to be the primary site of CR-allergen accumulation, but significant CR-allergen levels can also be found at other sites in the home. The MAb-based assays can be used for quantitation of environmental exposure to CR allergens.(ABSTRACT TRUNCATED AT 250 WORDS)

Allergens↗
PubMed1991

Identification, quantitation, and purification of cockroach allergens using monoclonal antibodies.

A panel of murine IgG monoclonal antibodies (MAbs) was raised against German cockroach (CR) (Blattella germanica) extract and selectively screened to identify MAb directed against allergen(s) recognized by IgE antibodies. Sera from 28 CR-allergic patients were used as sources of IgE antibodies to detect allergens "presented" by the MAb. Four clones (10A6, 3G12, 8F4, and 1D4) produced MAb to allergen(s) that bound IgE antibodies. Quantitative radioimmunoassays were used to compare levels of the MAb-defined allergens in CR extracts. MAb 10A6 reacted with a cross-reacting allergen that was detected in 9/14 CR species, including Blattella, Periplaneta, Blatta, Leucophea, and Supella spp, at concentrations of 100 to 10,000 U/ml. In contrast, MAb 3G12, 8F4, and 1D4 were Blattella specific. The allergen defined by MAb 8F4 was purified by MAb affinity chromatography and size-exclusion by high-performance liquid chromatography. It is a 36 kd heat-sensitive protein, isoelectric point, 5.2 to 5.4. Allergen 10A6 was partially purified by isoelectric focusing and high-performance liquid chromatography. It is a heat-stable, acidic protein (isoelectric point 3.15). Based on comparison of their properties with properties of previously described CR allergens, the allergens defined by MAb 10A6 and 8F4 have been provisionally designated Blattella germanica allergen I (Bla g I) and Blattella germanica allergen II (Bla g II), respectively. Assays of six commercial CR skin test extracts demonstrated a 200-fold difference in Bla g I levels (4.7 to 1085 U/ml) and only two extracts that contained detectable Bla g II (248 and 324 U/ml). The results demonstrate that MAb can be used to identify and define CR allergens and that the strategy of the use of MAb as a first step in allergen analysis and purification can be very effective, especially for poorly characterized allergen extracts.

Allergens↗
PubMed1991

Airborne dust mite allergens: comparison of group II allergens with group I mite allergen and cat-allergen Fel d I.

The form in which allergens become airborne is important because it may influence both symptoms caused by allergen exposure and methods used to reduce exposure. The group I allergens from dust mites only become airborne during disturbance and fall rapidly, which is in keeping with their being carried on fecal pellets. Their mean size is approximately 20 microns in diameter. By contrast, the cat-allergen Fel d I is airborne on particles varying from greater than 10 to less than 2 microns in diameter, some of which remain airborne even without disturbance. A second group of mite allergens, molecular weight 14,000, are equally important and are associated predominantly with mite bodies. With a monoclonal antibody-based assay and a cascade impactor, we have investigated the form in which group II mite allergens become airborne. The results reveal that these allergens only become airborne during disturbance and that they fall within 15 minutes. However, the mean size of particles carrying group II allergens appears to be slightly smaller than the mean size of particles carrying group I allergens. In addition, the quantities of group II allergen becoming airborne during disturbance (mean, 26 ng/m3) could not be explained by the quantity found in fecal particles. Thus, group II mite allergens become airborne in a form quite distinct from cat allergens and very similar to group I mite allergens; however, it appears unlikely that fecal particles are the main form in which group II allergens become airborne.

Allergens↗
PubMed1991

Exposure and sensitization to dust mite allergens among asthmatic children in São Paulo, Brazil.

A group of 20 mite allergic asthmatic children aged 6-12 years old, living in São Paulo, Brazil, was studied regarding their degree of sensitization to house dust mites and exposure to mite allergens in their homes. In 18 out of 20 houses at least one dust sample was obtained which contained greater than 10 micrograms Der p I/g of dust. The highest levels of Dermatophagoides pteronyssinus allergens, Der p I and Group II, were measured in bedding samples (geometric mean 38.4 and 36.6 micrograms/g, respectively), followed by bedroom floor, TV room and kitchen. Mite allergen levels in Brazilian houses were as high as those reported to be associated with sensitization and acute attacks of asthma in other parts of the world. In keeping with previous reports that D. farinae is rarely found in Brazil, Der f I was undetectable or found in very low levels (less than 0.5 micrograms/g). Levels of cat allergen Fel d I of greater than 8 micrograms/g of dust were obtained only in 2 houses only. Cockroach allergen Bla g I was detected in five out of 20 houses. Levels of IgE antibodies to D. pteronyssinus were greater than 200 RAST U/ml in 19 out of 20 children (geometric mean 1588 RAST U/ml). IgE antibodies to cat, cockroach, A. fumigatus, ragweed and rye grass pollens were undetectable or less than 80 RAST U/ml. IgE antibodies to the mite Blomia tropicalis were also measured, and levels greater than 200 RAST U/ml were observed in 13 out of 20 sera.(ABSTRACT TRUNCATED AT 250 WORDS)

Allergens↗
PubMed1991

Epidemiology of the relationship between exposure to indoor allergens and asthma.

A very high prevalence of immediate hypersensitivity to common indoor allergens can be demonstrated among children and young adults, with asthma. Recent progress in the immunochemistry of cat, dust mite and cockroach allergens has made it possible to measure exposure to these allergens and to start to define threshold levels of exposure which increase the risk of sensitization and symptomatic asthma. Indeed, it is already clear that exposure to greater than 2 micrograms group I dust mite allergen (or 100 mites) per gram of dust increases the risk of children developing sensitization and asthma. Furthermore, from studies on patients presenting to emergency rooms with asthma, it is clear that the risk of sensitization to allergens derived from cats or cockroaches or grass pollen is restricted to patients who are exposed to high levels of these allergens. Given the increasing morbidity and mortality of asthma it is clear that therapeutic efforts should be focused on identifying relevant allergens and advising patients about techniques for reducing exposure.

Air Pollution, Indoor↗
PubMed1991

Asthma in Tanta, Egypt: serologic analysis of total and specific IgE antibody levels and their relationship to parasite infection.

The relationship between asthma, IgE and parasite infection was compared in 68 randomly selected patients with asthma and 37 nonasthmatic controls living in Tanta, Egypt. Sera were assayed for total IgE and for IgE antibodies to inhaled allergens (mite, cat, cockroach, ryegrass, ragweed and 3 fungi) and to parasite antigens (Schistosoma mansoni and Brugia malayi). Parasite infection was determined by microscopic examination of stool specimens. Total IgE levels were significantly higher in patients with asthma (geometric mean 909 IU/ml), than in controls (geometric mean 145 IU/ml, p less than 0.001). The high IgE levels correlated with parasite infection and the presence of IgE antibodies to S. mansoni antigens, which were also elevated compared to controls. The prevalence of allergen-specific IgE antibodies among Egyptian asthmatics was low by comparison with 'Western' asthmatics, but nonetheless higher than among Egyptian controls. A radioallergosorbent test (RAST) values of greater than 40 U/ml to any allergen was found in 19/68 (28%) sera from the asthma group, as compared to only 1/37 (3%) sera from controls (p less than 0.001). The highest RAST values were to dust mite (Dermatophagoides pteronyssinus and D. farinae) allergens, followed by rye grass and ragweed allergens. The results suggest that in this area of Egypt, several factors may influence the development of asthma, including nonspecific activation of IgE and/or inflammatory mechanisms by helminth parasites and sensitisation to environmental allergens.

Adolescent↗