title: Sequential epitopes of Dermatophagoides farinae allergens identified using peptide microarray-based immunoassay.
abstract: House dust mites produce over 30 proteins proposed to induce immunoglobulin E (IgE) antibody production in patients. Continued identification of IgE-binding epitopes of these allergens is critical to advancing diagnosis and treatment of allergic disease. To identify possible sequential IgE-binding epitopes of the major- and mid-potency allergens from the house dust mite Dermatophagoides farinae by peptide microarray-based immunoassay, nucleotide sequences of D. farinae allergens (Der f) 1, 2, 4, 5, and 7 were used to generate overlapping peptides covering the full protein sequences minus signal peptides. Short peptides were printed onto microarray chips. Because asthma occurs as a symptom of mite allergy more commonly among children than adults, the peptide chips were exposed to sera pooled from six serum-positive pediatric patients with D. farinae hypersensitivity and six serum-negative control children for screening sequential IgE-binding epitopes by IgE immunolabeling. Higher-than-average immunolabel signal intensity was observed for 21 short peptides in the serum-positive group (P < 0.01). Due to sequence overlap, these 21 signals represented four fragments of Der f 1 (amino acid positions 46-53, 71-78, 99-110, 179-186), three fragments of Der f 2 (15-22, 80-89, 106-113), six fragments of Der f 4 (69-82, 107-116, 225-232, 261-268, 355-365, 483-496), one fragment of Der f 5 (102-109), and three fragments of Der f 7 (32-39, 52-64, 100-107). These findings not only demonstrate the utility of a peptide microarray immunoassay in identifying epitopes for these allergens, but also provide a foundation for future exploration of specific immunotherapies. © 2016 IUBMB Life, 68(10):792-798, 2016.
authors.affiliation: Department of Clinical Laboratory, The Third People's Hospital, Affiliated Yancheng Hospital, School of Medicine, Southeast University, Yancheng 224001, Jiangsu Province, People's Republic of China. ybcui1975@hotmail.Com.|Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Yancheng 224006, Jiangsu Province, People's Republic of China. ybcui1975@hotmail.Com.
authors.forename: Yubao
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Yancheng 224006, Jiangsu Province, People's Republic of China.
authors.forename: Feixiang
authors.initials: F
authors.lastname: Teng
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Yancheng 224006, Jiangsu Province, People's Republic of China.
authors.forename: LiLi
authors.initials: L
authors.lastname: Yu
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Yancheng 224006, Jiangsu Province, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Yancheng 224006, Jiangsu Province, People's Republic of China.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Yancheng 224006, Jiangsu Province, People's Republic of China.
authors.forename: Chengbo
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Laboratory Medicine, Yancheng Health Vocational & Technical College, Yancheng 224006, Jiangsu Province, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000737:chemistry / Q000276:immunology*; D001249:Asthma / Q000097:blood / Q000276:immunology*; D001665:Binding Sites; D002648:Child; D002675:Child, Preschool; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D005260:Female; D006801:Humans; D007118:Immunoassay; D007073:Immunoglobulin E / Q000097:blood / Q000737:chemistry; D008297:Male; D040081:Protein Array Analysis; D039981:Pyroglyphidae / Q000276:immunology*
chemical_list: D039741:Antigens, Dermatophagoides; D000939:Epitopes; D007073:Immunoglobulin E
keywords: epitopes; immunoglobulin E (IgE); mite allergens; peptide microarray