IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen
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Cockroaches have been recognized as a major cause of asthma. Bla g 4 is one of the most important German cockroach allergens. The aim of this study is to investigate IgE reactivity to the recombinant Bla g 4 (rBla g 4) in the sera of allergic patients and identify linear IgE binding epitope. For protein expression, full-length Bla g 4 (EF202172) was divided into 5 overlapping peptide fragments (E1: aa 1-100, E2: aa 34-77, E3: aa 74-117, E4: aa 114-156, and E5: aa 153-182). The full-length and 5 peptide fragments of Bla g 4 was generated by PCR and over-expressed in E. coli BL21 (DE3). The IgE binding reactivities of the full-length and peptide fragments were measured by ELISA using 32 serum samples of cockroach allergy. The sera of 8 patients (25%) reacted with rBla g 4. Four sera (100%) showed IgE-binding reactivity to fulllength and peptide fragment 4, and 2 sera (50%) reacted with peptide fragment 2. One (20%) serum reacted with peptide fragment 3. The results of ELISA using overlapping recombinant fragments indicated that the epitope region was located at amino acid sequences 34-73 and 78-113, and major IgE epitope of Bla g 4 was located at amino acid sequences 118- 152 of C-terminal. B-cell epitope analysis of German cockroach allergen Bla g 4 could contribute to the strategic development of more specific and potentially efficacious immunotherapy.
Abstract
Cockroaches have been recognized as a major cause of asthma. Bla g 4 is one of the most important German cockroach allergens. The aim of this study is to investigate IgE reactivity to the recombinant Bla g 4 (rBla g 4) in the sera of allergic patients and identify linear IgE binding epitope. For protein expression, full-length Bla g 4 (EF202172) was divided into 5 overlapping peptide fragments (E1: aa 1-100, E2: aa 34-77, E3: aa 74-117, E4: aa 114-156, and E5: aa 153-182). The full-length and 5 peptide fragments of Bla g 4 was generated by PCR and over-expressed in E. coli BL21 (DE3). The IgE binding reactivities of the full-length and peptide fragments were measured by ELISA using 32 serum samples of cockroach allergy. The sera of 8 patients (25%) reacted with rBla g 4. Four sera (100%) showed IgE-binding reactivity to fulllength and peptide fragment 4, and 2 sera (50%) reacted with peptide fragment 2. One (20%) serum reacted with peptide fragment 3. The results of ELISA using overlapping recombinant fragments indicated that the epitope region was located at amino acid sequences 34-73 and 78-113, and major IgE epitope of Bla g 4 was located at amino acid sequences 118- 152 of C-terminal. B-cell epitope analysis of German cockroach allergen Bla g 4 could contribute to the strategic development of more specific and potentially efficacious immunotherapy.
참고문헌 (25)
- Arruda LK, "Cloning of cockroach allergen, Bla g 4, identifies ligand binding proteins (or calycin) as a cause of IgE antibody responses", J BiolChem, 270, pp.196~201 (1995)
- Fan Y, "Tissue localization and regulation by juvenile hormone of human allergen Bla g 4 from the German cockroach", Blattella germanica.Insect Mol Biol, 14, pp.45~53 (2005)
- Flower DR, "The lipocalin protein family: structure and function", Biochem J, 318, pp.1~14 (1996)
- Virtanen T, "Important animal allergens are lipocalin protein: why are they allergenic?", Int Arch Allergy Immunol, 120, pp.247~258 (1999)
- Flower DR, "Structure and sequence relationships in the lipocalins and related proteins", Protein Sci, 2, pp.753~761 (1993)
- Vailes LD, "High-level expression of cockroach allergen, Bla g 4, in Pichia pastoris", J Allergy Clin Immunol, 101, pp.274~280 (1998)
- Jeong KY, "Recombinant allergens for diagnosis and immunotherapy of allergic disorders, with emphasis on cockroach allergy", Curr Protein Pept Sci, 7, pp.57~71 (2006)
- Larche M, "Peptide-based therapeutic vaccines for allergic and autoimmune disease", Nat Med, 11, pp.69~76 (2005)
- Jeong KY, "Sequence polymorphisms of major German cockroach allergen Bla g 1, Bla g 2, Bla g 4 and Bla g 5", Int Arch Allergy Immunol, 145, pp.1~8 (2008)
- Jeong KY, "Sequence diversity of the Bla g 4 cockroach allergen, homologous to lipocalins, from Blattella germanica", Int Arch Allergy Immunol, 148, pp.339~345 (2009)
- Satinover SM, "Specific IgE and IgG antibody-binding patterns to recombinant cockroach allergens", J Allergy Clin Immunol, 115, pp.803~809 (2005)
- Slater JE, "Biological potency of German cockroach allergen extracts determined in an inner city population", Clin Exp Allergy, 37, pp.1033~1039 (2007)
- Skerra A, "Lipocalins as a scaffold", Biochim Biophys Acta, 1482, pp.337~350 (2000)
- Olsson S, "Contribution of disulphide bonds to antigenicity of Lep d 2, the major allergen of the dust mite Lepidoglyphus destructor", Mol Immunol, 35, pp.1017~1023 (1998)
- Hakkaart GA, "Epitope mapping of the house-dust-mite allergen Der p 2 by means of site-directed mutagenesis", Allergy, 53, pp.165~172 (1998)
- Nishiyama C, "Analysis of the IgE-epitope of Der f 2, a major mite allergen, by in vitro mutagenesis", Mol Immunol, 32, pp.1021~1029 (1995)
- Kauppinen J, "Mutant derivatives of the main respiratory allergen of cow are less allergenic than the intact molecule", Clin Exp Allergy, 29, pp.989~996 (1999)
- Hales BJ, "Genetic variation of Der p 2 allergens: effects on T cell responses and immunoglobulin E binding", Clin Exp Allergy, 32, pp.1461~1467 (2002)
- Ferreira F, "Dissection of immunoglobulin E and T lymphocyte reactivity of isoforms of the major birch pollen allergen Bet v 1: potential use of hypoallergenic isoforms for immunotherapy", J Exp Med, 183, pp.599~609 (1996)
- Rouvinen J, "Search for the determinants of allergenicity in proteins of the lipocalin family", J Chromatogr B Biomed Sci Appl, 756, pp.199~206 (2001)
- Tan YW, "Structure of two major allergens, Bla g 4 and Per a 4, from cockroaches and their IgE binding epitopes", J Biol Chem, 284, pp.3148~3157 (2009)
- Selo I, "Allergy to bovine beta-lactoglobulin: specificity of human IgE to tryptic peptides", Clin Exp Allergy, 29, pp.1055~1063 (1999)
- Stanley JS, "Identification and mutational analysis of the immunodominant IgE binding epitopes of the major peanut allergen Ara h 2", Arch Biochem Biophys, 342, pp.244~253 (1997)
- Wang F, "Ana o1, a cashew (Ananardium occidental) allergen of the vicilin seed storage protein family", J Allergy Clin Immunol, 110, pp.160~166 (2002)
- Bredehorst R, "What establishes a protein as an allergen?", J Chromatogr B Biomed Sci, 756, pp.33~40 (2001)