Carbohydrates Contribute to the Interactions between Cockroach Allergen Bla g 2 and a Monoclonal Antibody

Li, Mi; Gustchina, Alla; Glesner, Jill; Wünschmann, Sabina; Vailes, Lisa D.; Chapman, Martin D.; Pomés, Anna; Wlodawer, Alexander

doi:10.4049/jimmunol.1002318

Cited by 37 publications

(56 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition to IgE-mediated allergic responses, these purified allergens can induce allergic inflammations that are not mediated by IgE interactions. For instance, the human beta-defensin 3 has been shown to bind Bla g 2 and could modify the ability of Bla g 2 to induce localized allergic inflammatory and mucosal responses (44, 45). Bla g 7 can potently induce T cell immunoglobulin mucin domain (TIM) 4 expression on dendritic cells (46), which plays a critical role in T cell proliferation and Th2 cell development (47, 48).…”

Section: Introductionmentioning

confidence: 99%

Cockroach allergen exposure and risk of asthma

Danh

Zhao

Gao

2016

Allergy

136

View full text Add to dashboard Cite

Cockroach sensitization is an important risk factor for the development of asthma. However, its underlying immune mechanisms and the genetic etiology for differences in allergic responses remain unclear. Cockroach allergens identification and their expression as biologically active recombinant proteins has provided a basis for studying the mechanisms regarding cockroach allergens induced allergic sensitization and asthma. Glycans in allergens may play a crucial role in the immunogenicity of allergic diseases. Protease-activated receptor (PAR)-2, Toll-like receptor (TLR), and C-type lectin receptors have been suggested to be important for the penetration of cockroach allergens through epithelial cells to mediate allergen uptake, dendritic cell maturation, antigen presenting cell (APC) function in T cell polarization, and cytokine production. Environmental pollutants, which often co-exist with the allergen, could synergistically elicit allergic inflammation, and aryl hydrocarbon receptor (AhR) activation and signaling may serve as a link between these two elements. Genetic factors may also play an important role in conferring the susceptibility to cockroach sensitization. Several genes have been associated with cockroach sensitization and asthma-related phenotypes. In this review, we will discuss the epidemiological evidence for cockroach allergen-induced asthma, cockroach allergens, the mechanisms regarding cockroach allergens induced innate immune responses, and the genetic basis for cockroach sensitization.

show abstract

Section: Introductionmentioning

confidence: 99%

Cockroach allergen exposure and risk of asthma

Danh

Zhao

Gao

2016

Allergy

136

View full text Add to dashboard Cite

show abstract

“…The most definitive approach to elucidating the structure of conformational epitope(s) of an allergen is by crystallographic analysis of allergen-antibody complexes [15,16,17,18,19,20]. This approach is not always available to investigators and can be laborious and time consuming.…”

Section: Introductionmentioning

confidence: 99%

Validation of a Phage Display and Computational Algorithm by Mapping a Conformational Epitope of Bla g 2

Tiwari

Negi

Braun

et al. 2011

Int Arch Allergy Immunol

Self Cite

View full text Add to dashboard Cite

Background: Bla g 2, one of the major cockroach allergens, induces a strong IgE response against conformational epitopes, and on reexposure, sensitized individuals often display symptoms of allergic rhinitis and asthma. The aim of the current study was to perform a test of the efficacy of a modified phage display screening, characterization of selected phages and an automated algorithm, EpiSearch, in locating an important conformational epitope. Methods: The monoclonal antibody 7C11, which partially inhibits the binding of patient IgE antibodies to Bla g 2, was used to screen a random peptide phage library. After 3 rounds of panning, 32 phage clones were isolated and the amino acid sequences of their peptides were determined. The relative affinity and specificity of the binding of these peptides to 7C11 were tested in ELISAs. The amino acid composition of these peptides was then matched with clusters of residues on the surface of the 3-dimensional (3D) structure of Bla g 2, using our EpiSearch algorithm. Results: The amino acid sequences of the peptides on selected phages differed at only one position, occupied by 1 of 2 negatively charged residues. The two 12-mer sequences bound to 7C11 with similar avidity and specificity. There was good concordance between the residues in the 3D clusters identified from our phage display/computational method with the co-crystal structural analysis. Conclusion: Conformational epitopes may be mapped through screening of clones from random peptide phage display libraries and EpiSearch.

show abstract

“…Our approach, which was based on a combination of x-ray crystallography and site-directed mutagenesis, showed the conformational nature of IgE epitopes (13)(14)(15)(16). The strategies for mutagenesis in this study were based on experimentally determined structures of Bla g 2 alone or in complex with antibodies (13)(14)(15). Therefore, the three-dimensional structure of the protein, essential for induction of immune responses by inhaled allergens, was an important consideration in our study.…”

Section: Discussionmentioning

confidence: 99%

“…There are no computational tools to accurately predict conformational epitopes, and several prediction methods available identify only linear B-cell epitopes (41)(42)(43). Our approach, which was based on a combination of x-ray crystallography and site-directed mutagenesis, showed the conformational nature of IgE epitopes (13)(14)(15)(16). The strategies for mutagenesis in this study were based on experimentally determined structures of Bla g 2 alone or in complex with antibodies (13)(14)(15).…”

Section: Discussionmentioning

confidence: 99%

“…The primary objective was to mutate residues involved in different IgE antibody-binding sites that had been either previously identified by mutagenesis of two Bla g 2 epitopes defined by x-ray crystallography or newly identified in this study (13)(14)(15)(16). Our findings show mechanisms of allergen-antibody interaction and reveal that Bla g 2 mutants displaying the same overall fold as the wild type allergen display reduced capacity to bind IgE but retain the ability to activate T-cells and modulate their response.…”

mentioning

confidence: 99%

See 1 more Smart Citation

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

Carbohydrates Contribute to the Interactions between Cockroach Allergen Bla g 2 and a Monoclonal Antibody

Cited by 37 publications

References 42 publications

Cockroach allergen exposure and risk of asthma

Cockroach allergen exposure and risk of asthma

Validation of a Phage Display and Computational Algorithm by Mapping a Conformational Epitope of Bla g 2

Antigenic Determinants of the Bilobal Cockroach Allergen Bla g 2

Contact Info

Product

Resources

About