Antigenic determinants of asthma-associated allergens for design of immunotherapy Project Summary/Abstract Allergic reactions to cockroach and dust mite are important health problems in the U.S. affecting up to 83% of asthmatic children in inner city areas and are a risk factor for emergency room admission with asthma. The main cockroach species in the U.S., Blattella germanica, produces Bla g 2 which induces sensitization at exposure levels 10-100 times lower than cat and mite allergens, and has the highest prevalence of sensitization among cockroach allergens (40-70%). Der p 1 and Der f 1 are cysteine proteases produced by the dust mites Dermatophagoides pteronyssinus and D. farinae, respectively. IgE sensitization to these Group 1 allergens is >80% among mite allergic patients. Proteolytic activity of Der p 1 may enhance IgE antibody production and contribute to lung inflammation in asthma but Bla g 2 is an inactive aspartic protease- homolog. The main goal of this project is to investigate the antigenic structure of proteolytic (mite Group 1) and non-proteolytic (Bla g 2) allergens associated with asthma. Allergens will be co-crystallized with monoclonal antibodies, and the key amino acids involved in antibody (IgE and mAb) binding will be identified. Alternatively, identification of IgE antibody binding epitopes will be performed by phage display technology.
The specific aims are: 1) mapping of antigenic determinants in Bla g 2 by crystallography;2) mapping of antigenic determinants of Group 1 mite allergens by crystallography and analysis of the structural basis for the cross-reactivity between Der p 1 and Der f 1;and 3) localization of the key amino acids involved in the antibody binding epitopes by site-directed mutagenesis studies and identification of hypoallergenic mutants with T cell reactivity that could be used for immunotherapy. IgE antibody binding to the epitope mutants will be analyzed by ELISA, multiplex array technology and cell mediator release assays. Mutants will be compared to select hypoallergenic forms for the design of vaccines for immunotherapy of mite and cockroach allergy.

Public Health Relevance

Cockroach and mite allergy is associated with the development of asthma, which affects 17 million people in the U.S. The antigenic determinants of Bla g 2 and Group 1 mite allergens will be identified to elucidate the importance of the intrinsic properties of these allergens on allergic disease. Hypoallergenic mutants will be produced and tested for IgE antibody binding and T cell proliferation, and the information obtained will facilitate a rational design of allergy vaccines.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI077653-04
Application #
8305481
Study Section
Hypersensitivity, Autoimmune, and Immune-mediated Diseases Study Section (HAI)
Program Officer
Minnicozzi, Michael
Project Start
2009-08-25
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2014-07-31
Support Year
4
Fiscal Year
2012
Total Cost
$429,847
Indirect Cost
$140,840
Name
Indoor Biotechnologies
Department
Type
DUNS #
007370633
City
Charlottesville
State
VA
Country
United States
Zip Code
22903
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Chapman, Martin D; Wuenschmann, Sabina; King, Eva et al. (2015) Technological Innovations for High-Throughput Approaches to In Vitro Allergy Diagnosis. Curr Allergy Asthma Rep 15:36
Osinski, Tomasz; Pomés, Anna; Majorek, Karolina A et al. (2015) Structural Analysis of Der p 1-Antibody Complexes and Comparison with Complexes of Proteins or Peptides with Monoclonal Antibodies. J Immunol 195:307-16

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