Specific IgE and IgG antibody-binding patterns to recombinant cockroach allergens

Satinover, S.M.; Reefer, A.J.; Pomés, Anna; Chapman, Martin D.; Platts-Mills, Thomas A.E.; Woodfolk, Judith A.

doi:10.1016/j.jaci.2005.01.018

Cited by 111 publications

(142 citation statements)

References 33 publications

(66 reference statements)

Supporting

Mentioning

134

Contrasting

Order By: Relevance

“…However, we recently found that natural Der f 1 did not dimerize in crystals. 4 Whereas it was not possible to conclude unambiguously that the dimeric state of mature Der p 1 observed in crystals is physiologically relevant, Bla g 2 is predicted to be a dimer under physiological conditions. The dimer interface between Bla g 2 monomers is extensive (1139 Å 2 per monomer), above the consensus cut-off value (ϳ850 Å 2 ) that allows distinction between contacts caused by crystal packing and real physiological contacts, but it is only 620 Å 2 for Der p 1 (27).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Crystal Structure of a Dimerized Cockroach Allergen Bla g 2 Complexed with a Monoclonal Antibody

Gustchina

Alexandratos

et al. 2008

Journal of Biological Chemistry

Self Cite

103

View full text Add to dashboard Cite

The crystal structure of a 1:1 complex between the German cockroach allergen Bla g 2 and the Fab fragment of a monoclonal antibody 7C11 was solved at 2.8-Å resolution. Bla g 2 binds to the antibody through four loops that include residues 60 -70, 83-86, 98 -100, and 129 -132. Cation-interactions exist between Lys-65, Arg-83, and Lys-132 in Bla g 2 and several tyrosines in 7C11. In the complex with Fab, Bla g 2 forms a dimer, which is stabilized by a quasi-four-helix bundle comprised of an ␣-helix and a helical turn from each allergen monomer, exhibiting a novel dimerization mode for an aspartic protease. A disulfide bridge between C51a and C113, unique to the aspartic protease family, connects the two helical elements within each Bla g 2 monomer, thus facilitating formation of the bundle. Mutation of these cysteines, as well as the residues Asn-52, Gln-110, and Ile-114, involved in hydrophobic interactions within the bundle, resulted in a protein that did not dimerize. The mutant proteins induced less ␤-hexosaminidase release from mast cells than the wild-type Bla g 2, suggesting a functional role of dimerization in allergenicity. Because 7C11 shares a binding epitope with IgE, the information gained by analysis of the crystal structure of its complex provided guidance for site-directed mutagenesis of the allergen epitope. We have now identified key residues involved in IgE antibody binding; this information will be useful for the design of vaccines for immunotherapy.Cockroach allergy is associated with the development of asthma and is a risk factor for emergency room admission of asthmatic patients, especially among inner city children living in low-income houses infested with cockroaches (1, 2). Cockroaches release allergens to the environment, which are carried by particles (5-40 m of diameter) that reach the lung by inhalation. Bla g 2 is one of the most important cockroach allergens, eliciting production of specific IgE in ϳ70% of cockroach-allergic patients at exposure levels that are 10 -100-fold lower than those from other common indoor allergens from dust mite and cat (3-5). Exposure to Bla g 2 results in cross-linking of IgE bound to the surface of mast cells or basophils from sensitized patients (i.e. in immunological terms, cross-linking refers to the non-covalent linkages between the allergen and two IgE molecules at the surface of mast cells or basophils), and induces release of potent mediators (histamine, leukotrienes, prostaglandins, etc.) of allergic reactions.We recently solved the crystal structure of Bla g 2, confirming that the overall fold of this allergen corresponds to that of pepsin-like aspartic proteases, and revealing structural elements that explain why it is enzymatically inactive (6, 7). Bla g 2 contains important amino acid substitutions in the area corresponding to the catalytic site. These modifications impair enzymatic function, and the allergen did not show proteolytic activity in standard in vitro assays using casein and hemoglobin as substrates (7). Bla g 2 belongs to a gro...

show abstract

Section: Discussionmentioning

confidence: 99%

“…allergens from dust mite and cat (3)(4)(5). Exposure to Bla g 2 results in cross-linking of IgE bound to the surface of mast cells or basophils from sensitized patients (i.e.…”

mentioning

confidence: 99%

Crystal Structure of a Dimerized Cockroach Allergen Bla g 2 Complexed with a Monoclonal Antibody

Gustchina

Alexandratos

et al. 2008

Journal of Biological Chemistry

Self Cite

103

View full text Add to dashboard Cite

show abstract

“…Sensitisation and exposure data suggest that Bla g 2 induces strong IgE response at exposure levels that are often 10 to 100 times lower than levels of other common indoor allergens (9,13,42). However, group 1 allergens (Bla g 1 and Per a 1 produced by B. germanica and P. americana, respectively) share homology in their structure and signifi cant crossreactivity (7,8,11,12).…”

Section: Statisticsmentioning

confidence: 99%

“…While B. germanica is common in the northern hemisphere, especially in the United States and Europe, P. americana is predominant in tropical countries such as Taiwan and Brazil, especially in food related areas (7). B. germanica contains multiple allergens (Bla g 1, Bla g Bd90K, Bla g 2, Bla g 4, Bla g 5, Bla g 6, Bla g 7), of which Bla g 1 and Bla g 2 have been produced for immunoassays (7)(8)(9). Bla g 1 belongs to cross-reacting group 1 allergens (10,11).…”

mentioning

confidence: 99%

Bla g 1 Allergen Levels in Zagreb Area Household Dust

Prester¹,

Macan²

2011

Archives of Industrial Hygiene and Toxicology

View full text Add to dashboard Cite

Cockroach allergy is a health problem in many parts of the world. In urban environments, indoor exposure to cockroach allergens involves a risk of asthma. The aim of this study was to measure the mass fraction of Bla g 1, a major allergen of the German cockroach (Blatella germanica) in 30 house samples, collected at random from Zagreb area households, Croatia. Dust samples were collected on cellulose fi lters by vacuuming living rooms fl oors. After extraction, Bla g 1 was detected using the commercial enzyme-linked immunosorbent assay (ELISA). Only four of the thirty households had detectable Bla g 1 levels, and only in one was its concentration higher than 2.0 U g -1 , the threshold associated with sensitisation. The Bla g 1 ELISA proved highly sensitive, with the detection limit of 0.12 U g -1. The within-and between-assay imprecision was 8.9 % and 14.4 %, respectively, and accuracy 85 % to 120 %. Low Bla g 1 levels in the household dust support previously reported low prevalence of skin sensitisation to B. germanica among Zagreb residents. Further monitoring should reveal if there are differences in cockroach allergen exposure and sensitisation between households from other geographic areas in Croatia.

show abstract

“…Natural or recombinant cockroach allergens have been used to evaluate their importance in allergic disease. Of all B. germanica allergens, patients show the highest prevalence of sensitization (ϳ55-70%) to Bla g 2, and this major allergen was therefore selected as the main focus of our studies (10,11). A Bla g 2 homolog from P. americana also showed a high prevalence of sensitization (81%) among patients with persistent asthma and rhinitis in Taiwan (7).…”

mentioning

confidence: 99%

Antigenic Determinants of the Bilobal Cockroach Allergen Bla g 2

Woodfolk¹,

Glesner²,

Wright³

et al. 2016

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Bla g 2 is a major indoor cockroach allergen associated with the development of asthma. Antigenic determinants on Bla g 2 were analyzed by mutagenesis based on the structure of the allergen alone and in complex with monoclonal antibodies that interfere with IgE antibody binding. The structural analysis revealed mechanisms of allergen-antibody recognition through cation-interactions. Single and multiple Bla g 2 mutants were expressed in Pichia pastoris and purified. The triple mutant K132A/K251A/F162Y showed an ϳ100-fold reduced capacity to bind IgE, while preserving the native molecular fold, as proven by x-ray crystallography. This mutant was still able to induce mast cell release. T-cell responses were assessed by analyzing Th1/Th2 cytokine production and the CD4 ؉ T-cell phenotype in peripheral blood mononuclear cell cultures. Although T-cell activating capacity was similar for the KKF mutant and Bla g 2 based on CD25 expression, the KKF mutant was a weaker inducer of the Th2 cytokine IL-13. Furthermore, this mutant induced IL-10 from a non-T-cell source at higher levels that those induced by Bla g 2. Our findings demonstrate that a rational design of site-directed mutagenesis was effective in producing a mutant with only 3 amino acid substitutions that maintained the same fold as wild type Bla g 2. These residues, which were involved in IgE antibody binding, endowed Bla g 2 with a T-cell modulatory capacity. The antigenic analysis of Bla g 2 will be useful for the subsequent development of recombinant allergen vaccines.

show abstract

Specific IgE and IgG antibody-binding patterns to recombinant cockroach allergens

Cited by 111 publications

References 33 publications

Crystal Structure of a Dimerized Cockroach Allergen Bla g 2 Complexed with a Monoclonal Antibody

Crystal Structure of a Dimerized Cockroach Allergen Bla g 2 Complexed with a Monoclonal Antibody

Bla g 1 Allergen Levels in Zagreb Area Household Dust

Antigenic Determinants of the Bilobal Cockroach Allergen Bla g 2

Contact Info

Product

Resources

About