Preparation and identification of Per a 5 as a novel American cockroach allergen.
Glutathione S-transferase (GST) from various arthropods can elicit allergic reactions. In the present study, Per a 5, a GST, was cloned from American cockroach (CR) and expressed in both baculovirus-infected insect cell (iPer a 5) and E. coli expression (bPer a 5) systems. The secondary structures were predicted to be 45.93 and 8.69% of α-helix β-sheets in iPer a 5 and 42.54 and 8.49% of α-helix and β-sheets in bPer a 5, respectively. It is found that 4 out of 16 (25%) sera from American CR allergy patients reacted to both bPer a 9 and iPer a 9 as assessed by ELISA and Western blotting analysis, confirming that Per a 5 is not a major allergen of American CR. Induction of upregulated expression of CD63 and CCR3 on passively sensitized human basophils (sera from American CR allergy patients) by approximately up to 4.5- and 3.2-fold indicates that iPer a 5 and bPer a 5 are functionally active. Recombinant Per a 5 (rPer a 5) should be a useful tool for studying and understanding the role of Per a 5 in CR allergy.
Allergy and Clinical Immunology Research Centre, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121001, China ; Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
Ji-Fu
0000-0001-6412-3251
JF
Wei
Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
Haiwei
0000-0001-8984-5007
H
Yang
Allergy and Clinical Immunology Research Centre, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121001, China.
Dongning
D
Li
Johns Hopkins Asthma & Allergy Center, Baltimore, MD 21224, USA.
Peisong
P
Gao
Allergy and Clinical Immunology Research Centre, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121001, China ; Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, China.
Shaoheng
S
He
D000485:Allergens / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D001483:Base Sequence; D001491:Basophils / Q000378:metabolism*; D002460:Cell Line; D003058:Cockroaches; D004926:Escherichia coli / Q000378:metabolism; D005786:Gene Expression Regulation; D005982:Glutathione Transferase / Q000276:immunology*; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D019476:Insect Proteins / Q000276:immunology*; D007313:Insecta; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D054397:Receptors, CCR3 / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology; D065631:Rhinitis, Allergic / Q000097:blood / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D060149:Tetraspanin 30 / Q000378:metabolism
D000485:Allergens; C515350:CCR3 protein, human; D019476:Insect Proteins; C000593531:Per a 5 allergen, Periplaneta americana; D054397:Receptors, CCR3; D011994:Recombinant Proteins; D060149:Tetraspanin 30; D005982:Glutathione Transferase