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Identification and characterization of a novel allergen from Blomia tropicalis: Blo t 21  Yun Feng Gao, BEng, De Yun Wang, MD, PhD, Tan Ching Ong, PhD,

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Presentation on theme: "Identification and characterization of a novel allergen from Blomia tropicalis: Blo t 21  Yun Feng Gao, BEng, De Yun Wang, MD, PhD, Tan Ching Ong, PhD,"— Presentation transcript:

1 Identification and characterization of a novel allergen from Blomia tropicalis: Blo t 21 
Yun Feng Gao, BEng, De Yun Wang, MD, PhD, Tan Ching Ong, PhD, Su Ling Tay, BSc(Hons), Kwong Hsia Yap, BSc, Fook Tim Chew, PhD  Journal of Allergy and Clinical Immunology  Volume 120, Issue 1, Pages (July 2007) DOI: /j.jaci Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Comparison between Blo t 21 protein sequence with Der p 21 and group 5 allergens. Identical and similar sequences were shaded with black and gray, respectively. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Southern blot analysis of Blo t 21 and Blo t 5 genes in the mite genome. A, Hybridization with a Blo t 5 probe. B, Hybridization with a Blo t 21 probe. In both, genomic DNA was digested with the restriction enzymes HindIII and EcoRV, respectively. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Expression of rBlo t 21 and rBlo t 5 allergens and their far UV CD spectra. A, Protein profile of rBlo t 21 and rBlo t 5 separated on a 15% SDS-PAGE gel. B, Far UV CD spectra of rBlo t 21 and rBlo t 5 proteins in solution. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Detection of nBlo t 5 and nBlo t 21 allergens in the crude extract. A, SDS-PAGE gel image of proteins loaded onto the gel. Immunoblotting images with anti-Blo t 5 (B) and anti-Blo t 21 polyclonal antibodies (C) and PBS (as a negative control; D). A nonspecific band (∗) was detected in all 3 immunoblotting membranes. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 Immunostaining of Blo t 21 and Blo t 5 in paraffin-embedded sections of B tropicalis. A, Probe with anti-Blo t 21 polyclonal antibody in longitudinal and sagittal mite sections. B, Probe with anti-Blo t 5 polyclonal antibody. Amg, Anterior midgut; Hg, hindgut. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Allergenicity of Blo t 21 and Blo t 5. A, Specific IgE levels assessed by means of ELISA in sera of 43 patients with allergic rhinitis. B, Specific IgE levels assessed by means of dot blot immunoassay in sera of 494 atopic individuals. Gray lines indicate the negative cutoff point. C, SPT responses among the 43 patients with allergic rhinitis. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 7 Dose-response competitive ELISA assay evaluating the IgE cross-reactivity of Blo t 21 and Blo t 5. Three individual sera were preabsorbed with 10-fold serial dilution of inhibitors and then assayed by using allergen-specific IgE ELISA. A, Blo t 21 immobilized on the solid phase of ELISA plates. B, Blo t 5 immobilized on the solid phase of ELISA plates. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2007 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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