title: Molecular cloning, expression, IgE binding activities and in silico epitope prediction of Per a 9 allergens of the American cockroach.
abstract: Per a 9 is a major allergen of the American cockroach (CR), which has been recognized as an important cause of imunoglobulin E-mediated type I hypersensitivity worldwide. However, it is not neasy to obtain a substantial quantity of this allergen for use in functional studies. In the present study, the Per a 9 gene was cloned and expressed in Escherichia coli (E. coli) systems. It was found that 13/16 (81.3%) of the sera from patients with allergies caused by the American CR reacted to Per a 9, as assessed by enzyme-linked immunosorbent assay, confirming that Per a 9 is a major allergen of CR. The induction of the expression of CD63 and CCR3 in passively sensitized basophils (from sera of patients with allergies caused by the American CR) by approximately 4.2-fold indicated that recombinant Per a 9 was functionally active. Three immunoinformatics tools, including the DNAStar Protean system, Bioinformatics Predicted Antigenic Peptides (BPAP) system and the BepiPred 1.0 server were used to predict the potential B cell epitopes, while Net-MHCIIpan-2.0 and NetMHCII-2.2 were used to predict the T cell epitopes of Per a 9. As a result, we predicted 11 peptides (23-28, 39-46, 58-64, 91-118, 131-136, 145-154, 159-165, 176-183, 290-299, 309-320 and 338-344) as potential B cell linear epitopes. In T cell prediction, the Per a 9 allergen was predicted to have 5 potential T cell epitope sequences, 119-127, 194-202, 210-218, 239-250 and 279-290. The findings of our study may prove to be useful in the development of peptide-based vaccines to combat CR-induced allergies.
authors.affiliation: Allergy and Clinical Immunology Research Centre, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China.
authors.forename: Haiwei
authors.initials: H
authors.lastname: Yang
authors.affiliation: Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.affiliation: Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Jin
authors.affiliation: Department of Respiratory Medicine, General Hospital of Shenyang Military Region, PLA Cancer Center, Shenyang, Liaoning 110840, P.R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Xie
authors.affiliation: Allergy and Clinical Immunology Research Centre, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China.
authors.forename: Shaoheng
authors.initials: S
authors.lastname: He
authors.affiliation: Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.
authors.forename: Ji-Fu
authors.initials: JF
authors.lastname: Wei
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D001122:Arginine Kinase / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D001483:Base Sequence; D001491:Basophils / Q000276:immunology; D001665:Binding Sites; D003001:Cloning, Molecular; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology*; D015870:Gene Expression; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology*; D008958:Models, Molecular; D010526:Periplaneta / Q000235:genetics* / Q000276:immunology*; D010802:Phylogeny; D011487:Protein Conformation; D011994:Recombinant Proteins; D065631:Rhinitis, Allergic / Q000097:blood / Q000276:immunology
chemical_list: D000485:Allergens; D018985:Epitopes, B-Lymphocyte; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D001122:Arginine Kinase; C553356:Per a 9 allergen, Periplaneta americana