A two-site monoclonal antibody ELISA for the quantification of the major Dermatophagoides spp. allergens, Der p I and Der f I

doi:10.1016/0022-1759(89)90010-0

Journal of Immunological Methods

Volume 118, Issue 2, 31 March 1989, Pages 227-235

https://doi.org/10.1016/0022-1759(89)90010-0 Get rights and content

Abstract

A two-site monoclonal antibody (Mab) ELISA was developed to measure the Group I allergens from Dermatophagoides spp., Der p I from D. pteronyssinus and Der f I from D. farinae. Species-specific Mabs were used to coat microtiter plates which were then incubated with allergen or house dust extracts. Bound allergen was detected using a biotinylated Mab which recognized a common epitope on both Der p I and Der f I, followed by the addition of streptavidin-peroxidase and ABTS/H₂O₂ substrate. The assay had low non-specific binding (∼ 0.08 absorbance units) and had a sensitivity of 5 ng/nl for aqueous allergen extracts (equivalent to 0.1 μg allergen/g dust). 53 dust samples were assayed using the Mab ELISA and an RIA previously described using ¹²⁵I-labelled Mab. The results showed a very good quantitative correlation between the assays (r = 0.96, p < 0.001 for Derp I; r = 0.92, P < 0.001 for Derf I). A further 132 dust samples from a different geographical areas were also assayed by both methods and gave correlation coefficients of 0.90 (P < 0.001) and (P < 0.001) for Der p I and Der f I, respectively. The Mab ELISA will be useful in epidemiological studies of allergic asthma, both in the assessment of levels of dust mite allergen present in houses and the efficacy of allergen avoidance regimes.

References (26)

M.D. Chapman et al.
Monoclonal immunoassays for major dust mite (Dermatophagoides) allergens, Der p I and Der f I, and quantitative analysis of the allergen content of mite and house dust extracts
J. Allergy Clin. Immunol.
(1987)
A.W. Ford et al.
A collaborative study on the first international standard of Dermatophagoides pteronyssinus (house dust mite) extract
J. Allergy Clin. Immunol.
(1985)
P. Lind
Purification and partial characterization of two major allergens from the house dust mite Dermatophagoides pteronyssinus
J. Allergy Clin. Immunol.
(1985)
M. Lombardero et al.
Monoclonal antibody based radioimmunoassay for the quantitation of the main cat salivary allergen (Fel d I or Cat-1)
J. Immunol. Methods
(1988)
T.A.E. Platts-Mills et al.
Dust mites: immunology, allergic disease and environmental control
J. Allergy Clin. Immunol.
(1987)
T.A.E. Platts-Mills et al.
Cross-reacting and species-specific determinants on a major allergen from Dermatophagoides pteronyssinus and D. farinae: development of a radioimmunoassay for antigen P₁ equivalent in house dust and mite extracts
J. Allergy Clin. Immunol.
(1986)
M.C. Swanson et al.
An immunochemical approach to indoor aeroallergen quantitation with a new volumetric air sampler: studies with mite, roach, cat, mouse and guinea pig antigens
J. Allergy Clin. Immunol.
(1985)
R. Voorhorst et al.
The house dust mite (Dermatophagoides pteronyssinus) and the allergens it produces: identity with the house dust allergen
J. Allergy
(1967)
J.J. Wadsley et al.
The effect of pH on the aggregation of biotinylated antibodies and on the signal-to-noise observed in immunoassays utilizing biotinylated antibodies
J. Immunol. Methods
(1987)
L.G. Arlian et al.
The prevalence of house dust mites, Dermatophagoides spp., and associated environmental conditions in homes in Ohio
J. Allergy Clin. Immunol.
(1982)

M.D. Chapman et al.

Purification and characterization of the major allergen from Dermatophagoides pteronyssinus — antigen P₁

J. Immunol.

(1980)

M.D. Chapman et al.

Recognition of two Dermatophagoides pteronyssinus-specific epitopes on antigen P₁ by using monoclonal antibodies: binding to each epitope can be inhibited by sera from dust-mite allergic patients

J. Immunol.

(1984)

M.D. Chapman et al.

Epitope mapping of two major inhalant allergens, Der p I and Der f I, from mites of the genus Dermatophagoides

J. Immunol.

(1987)

Cited by (393)

Efficacy of House Dust Mite Sublingual Immunotherapy in Patients with Atopic Dermatitis: A Randomized, Double-Blind, Placebo-Controlled Trial
2022, Journal of Allergy and Clinical Immunology: In Practice
Sensitization to house dust mites (HDMs) is frequent in patients with atopic dermatitis.
To investigate the efficacy of sublingual immunotherapy (SLIT) with Dermatophagoides pteronyssinus extract in patients with atopic dermatitis sensitized to HDM.
In this randomized, double-blind, placebo-controlled trial, we enrolled 91 patients 3 years or older, with SCORing Atopic Dermatitis (SCORAD) score greater than or equal to 15 and positive skin test result and/or IgE to D pteronyssinus. Patients were stratified according to age (<12 and ≥12 years) to receive HDM SLIT or placebo for 18 months. Primary outcome was a greater than or equal to 15-point decrease in SCORAD score. Secondary outcomes were decreases in SCORAD and objective SCORAD, Eczema Area and Severity Index, visual analog scale for symptoms, and pruritus scale scores; Investigator’s Global Assessment 0/1; and decrease greater than or equal to 4 points in Dermatology Life Quality Index. Background therapy was maintained.
A total of 66 patients completed the study (35 HDM SLIT, 31 placebo). After 18 months, 74.2% and 58% of patients in the HDM SLIT group and the placebo group, respectively, showed greater than or equal to 15-point decrease in SCORAD score (relative risk, 1.28; 95% CI, 0.89-1.83). Significant SCORAD score decreases from baseline of 55.6% and 34.5% in HDM SLIT and placebo groups (mean difference, 20.4; 95% CI, 3.89-37.3), significant objective SCORAD score decreases of 56.8% and 34.9% in HDM SLIT and placebo groups (mean difference, 21.3; 95% CI, 0.66-41.81), and more patients with Investigator’s Global Assessment 0/1 in the HDM SLIT group as compared with the placebo group (14 of 35 vs 5 of 31; relative risk, 2.63; 95% CI, 1.09-6.39) were observed at 18 months.
Our results suggest that HDM SLIT may be effective in HDM-sensitized patients as an add-on treatment for atopic dermatitis.
Presence of the house dust mite allergen in the gastrointestinal tract of dogs with chronic enteropathy: A potential inducer of interleukin-1β
2020, Veterinary Immunology and Immunopathology
Citation Excerpt :
The amount of Der p 1 protein was measured using a Der p 1 ELISA kit 5H8/4C1 (Indoor Biotechnologies) as previously described (Macchiaverni et al., 2014; Tulic et al., 2016). This ELISA kit specifically quantified Der p 1 using two different monoclonal antibodies (5H8 and 4C1) that were validated previously (Luczynska et al., 1989). Der p 1 levels in faeces (pg/g) were calculated by the following formula: the concentration of Der p 1 in the faecal solution (pg/mL) × volume of the solution (mL) ÷ weight of faecal samples used for faecal solution extraction (g).
House dust mite (HDM) is an environmental allergen ubiquitously present indoors, causing allergic inflammation in dogs. However, it is unclear whether HDM allergens can be detected in the gastrointestinal (GI) tract of dogs. In addition, although expression of interleukin (IL)-1β is increased in the intestinal mucosa of dogs with chronic enteropathy (CE), the role of HDM allergens in the production of IL-1β has not been evaluated. The objectives of this study were to determine the presence of HDM allergens in the GI tract of dogs and to elucidate the effect of HDM on IL-1β expression in canine macrophages. HDM allergen, Dermatophagoides pteronyssinus (Der p) 1, was quantified in the gastric and duodenal fluids and the duodenal and colonic mucosae of dogs with CE and healthy laboratory dogs, and faeces of dogs with CE, healthy laboratory dogs and healthy client-owned dogs. Gene expression and protein levels of IL-1β were measured in HDM-stimulated canine peripheral macrophages from healthy laboratory dogs. Der p 1 was detected in the gastric and duodenal fluids of dogs with CE and healthy laboratory dogs, and faeces of all dogs examined. Der p 1 levels in the duodenal and colonic mucosae were significantly higher in dogs with CE than in healthy laboratory dogs. HDM increased both gene expression and protein levels of IL-1β in canine macrophages. These findings demonstrate the presence of HDM allergens in the GI tract of dogs and the possible involvement of HDM allergens in the pathogenesis of CE by promoting IL-1β expression in macrophages.
Diagnosis of allergy in asthma
2020, Revue des Maladies Respiratoires
L’allergie est une réaction d’hypersensibilité provoquée par des mécanismes immunologiques. Dans l’asthme, l’allergie joue un rôle complexe et il s’agit le plus souvent d’un mécanisme médié par les IgE. Elle doit être recherchée au cours de l’évaluation d’un asthme mais la mise en évidence d’une sensibilisation IgE médiée ne signifie pas de factoque l’allergène joue un rôle dans la pathologie asthmatique. Afin de faire la part des choses, il faut évaluer la pertinence clinique de la sensibilisation. Dans cet article, nous décrivons les moyens actuellement disponibles pour rechercher une sensibilisation IgE médiée chez l’asthmatique mais également les moyens d’évaluer l’exposition allergénique environnementale et son rôle dans l’asthme.
Allergy is a hypersensitivity reaction induced by immunological mechanisms. In asthma, allergy has a complex role and is usually IgE mediated. Allergy must be evaluated during the work up but evidence of IgE sensitivity does not mean that allergens play a role in the pathophysiology of the disease. The clinical relevance of the sensitivity has to be considered. This paper describes current available tools to screen for IgE sensitivity, allergen exposure and their role in asthma.
Genome-wide interaction study of dust mite allergen on lung function in children with asthma
2017, Journal of Allergy and Clinical Immunology
Childhood asthma is likely the result of gene-by-environment (G × E) interactions. Dust mite is a known risk factor for asthma morbidity. Yet, there have been no genome-wide G × E studies of dust mite allergen on asthma-related phenotypes.
We sought to identify genetic variants whose effects on lung function in children with asthma are modified by the level of dust mite allergen exposure.
A genome-wide interaction analysis of dust mite allergen level and lung function was performed in a cohort of Puerto Rican children with asthma (Puerto Rico Genetics of Asthma and Lifestyle [PRGOAL]). Replication was attempted in 2 independent cohorts, the Childhood Asthma Management Program (CAMP) and the Genetics of Asthma in Costa Rica Study.
Single nucleotide polymorphism (SNP) rs117902240 showed a significant interaction effect on FEV₁ with dust mite allergen level in PRGOAL (interaction P = 3.1 × 10⁻⁸), and replicated in the same direction in CAMP white children and CAMP Hispanic children (combined interaction P = .0065 for replication cohorts and 7.4 × 10⁻⁹ for all cohorts). Rs117902240 was positively associated with FEV₁ in children exposed to low dust mite allergen levels, but negatively associated with FEV₁ in children exposed to high levels. This SNP is on chromosome 8q24, adjacent to a binding site for CCAAT/enhancer-binding protein beta, a transcription factor that forms part of the IL-17 signaling pathway. None of the SNPs identified for FEV₁/forced vital capacity replicated in the independent cohorts.
Dust mite allergen exposure modifies the estimated effect of rs117902240 on FEV₁ in children with asthma. Analysis of existing data suggests that this SNP may have transcription factor regulatory functions.
Cockroach allergen exposure and plasma cytokines among children in a tropical environment
2017, Annals of Allergy, Asthma and Immunology
Little is known about the effects of socioeconomic status or cockroach allergen on immune responses in school-age children, particularly in tropical environments.
To examine whether cockroach allergen and/or socioeconomic status is associated with plasma cytokine levels in Puerto Rican children.
This was a cross-sectional study of 532 children (6–14 years old) with (n = 272) and without (n = 260) asthma in San Juan (Puerto Rico). House dust allergens (cockroach [Bla g 2], dust mite [Der p 1], cat dander [Fel d 1], dog dander [Can f 1], and mouse urinary protein [Mus m 1]) were quantified using monoclonal antibody arrays. A panel of 14 cytokines (interleukin [IL]-1β, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon-γ, and tumor necrosis factor-α) was measured in plasma samples. Low household income was defined as less than $15,000 per year (below the median income for Puerto Rico in 2008–2009). Linear regression was used for the analysis of cockroach allergen and plasma cytokines.
In a multivariable analysis adjusting for low income and other allergen levels, cockroach allergen was significantly associated with decreased IL-17A and with increased levels of 8 cytokines (IL-4, IL-10, IL-17F, IL-21, IL-25, IL-31, interferon-γ, and tumor necrosis factor-α). After stratifying this analysis by cockroach allergy (ie, having a cockroach positive immunoglobulin E reaction), our findings remained largely unchanged for children sensitized to cockroach but became weaker and statistically nonsignificant for non-sensitized children.
Cockroach allergen has broad effects on adaptive immune responses in school-age children in a tropical environment, particularly in those sensitized to cockroach.
Relevance of specific IgE antibody titer to the prevalence, severity, and persistence of asthma among 19-year-olds in northern Sweden
2016, Journal of Allergy and Clinical Immunology
Although sensitization to indoor allergens is strongly associated with asthma, there are questions as to how this relates to asthma symptoms.
We sought to study the relevance of IgE antibodies to cat and dog allergens in an area in which (1) the climate discourages cockroach, fungal, and mite growth and (2) dander allergens are known to be present in schools and houses without animals.
IgE to 8 allergens was tested in 963 sera from a population-based study on 19-year-olds, and associations with asthma symptoms, diagnosis, and treatment were examined. In positive sera IgE to specific cat and dog allergens was also assayed.
IgE specific for animal dander had the highest prevalence and strongest relationship to asthma diagnosis. Furthermore, asthma severity, as judged by the frequency of symptoms and use of treatment, was directly associated with the titer of IgE antibodies to animal dander. Among the 103 subjects who had current asthma at age 19 years, 50 had asthma before age 12 years. Among those 50, the odds ratios for asthma related to any IgE antibodies to animal dander or high-titer IgE antibodies (≥17.5 IU/mL) were 9.2 (95% CI, 4.9-17) and 13 (95% CI, 6.9-25), respectively. In multivariable analysis IgE antibodies to Fel d 1 and Can f 5 were each associated with current asthma.
High-titer IgE antibodies to cat and dog allergens were strongly associated with the diagnosis, severity, and persistence of asthma; however, a large proportion of patients with current asthma did not live in a house with a cat or dog.

View all citing articles on Scopus

View full text

Article preview

Journal of Immunological Methods

Abstract

References (26)

Monoclonal immunoassays for major dust mite (Dermatophagoides) allergens, Der p I and Der f I, and quantitative analysis of the allergen content of mite and house dust extracts

J. Allergy Clin. Immunol.

A collaborative study on the first international standard of Dermatophagoides pteronyssinus (house dust mite) extract

J. Allergy Clin. Immunol.

Purification and partial characterization of two major allergens from the house dust mite Dermatophagoides pteronyssinus

J. Allergy Clin. Immunol.

Monoclonal antibody based radioimmunoassay for the quantitation of the main cat salivary allergen (Fel d I or Cat-1)

J. Immunol. Methods

Dust mites: immunology, allergic disease and environmental control

J. Allergy Clin. Immunol.

Cross-reacting and species-specific determinants on a major allergen from Dermatophagoides pteronyssinus and D. farinae: development of a radioimmunoassay for antigen P₁ equivalent in house dust and mite extracts

J. Allergy Clin. Immunol.

An immunochemical approach to indoor aeroallergen quantitation with a new volumetric air sampler: studies with mite, roach, cat, mouse and guinea pig antigens

J. Allergy Clin. Immunol.

The house dust mite (Dermatophagoides pteronyssinus) and the allergens it produces: identity with the house dust allergen

J. Allergy

The effect of pH on the aggregation of biotinylated antibodies and on the signal-to-noise observed in immunoassays utilizing biotinylated antibodies

J. Immunol. Methods

The prevalence of house dust mites, Dermatophagoides spp., and associated environmental conditions in homes in Ohio

J. Allergy Clin. Immunol.

Purification and characterization of the major allergen from Dermatophagoides pteronyssinus — antigen P₁

J. Immunol.

Recognition of two Dermatophagoides pteronyssinus-specific epitopes on antigen P₁ by using monoclonal antibodies: binding to each epitope can be inhibited by sera from dust-mite allergic patients

J. Immunol.

Epitope mapping of two major inhalant allergens, Der p I and Der f I, from mites of the genus Dermatophagoides

J. Immunol.

Cited by (393)

Efficacy of House Dust Mite Sublingual Immunotherapy in Patients with Atopic Dermatitis: A Randomized, Double-Blind, Placebo-Controlled Trial

Presence of the house dust mite allergen in the gastrointestinal tract of dogs with chronic enteropathy: A potential inducer of interleukin-1β

Diagnosis of allergy in asthma

Genome-wide interaction study of dust mite allergen on lung function in children with asthma

Cockroach allergen exposure and plasma cytokines among children in a tropical environment

Relevance of specific IgE antibody titer to the prevalence, severity, and persistence of asthma among 19-year-olds in northern Sweden

A two-site monoclonal antibody ELISA for the quantification of the major Dermatophagoides spp. allergens, Der p I and Der f I

Abstract

J. Allergy Clin. Immunol.

J. Allergy Clin. Immunol.

J. Allergy Clin. Immunol.

J. Immunol. Methods

J. Allergy Clin. Immunol.

J. Allergy Clin. Immunol.

J. Allergy Clin. Immunol.

J. Allergy

J. Immunol. Methods

The prevalence of house dust mites, Dermatophagoides spp., and associated environmental conditions in homes in Ohio

J. Allergy Clin. Immunol.

Purification and characterization of the major allergen from Dermatophagoides pteronyssinus — antigen P1

J. Immunol.

Recognition of two Dermatophagoides pteronyssinus-specific epitopes on antigen P1 by using monoclonal antibodies: binding to each epitope can be inhibited by sera from dust-mite allergic patients

J. Immunol.

Epitope mapping of two major inhalant allergens, Der p I and Der f I, from mites of the genus Dermatophagoides

J. Immunol.

Purification and characterization of the major allergen from Dermatophagoides pteronyssinus — antigen P₁

Recognition of two Dermatophagoides pteronyssinus-specific epitopes on antigen P₁ by using monoclonal antibodies: binding to each epitope can be inhibited by sera from dust-mite allergic patients