title: Il17A autoantibody induced by recombinant protein Ag85A-Il17A.
abstract: IL17A is a widely studied cytokine which plays an important role in autoimmune diseases as well as asthma. In this study, murine IL17a gene fragment was inserted into pET28a-Ag85A and it was expressed as a fusion protein named Ag85A-IL17A. Recombinant protein Ag85A-IL17A was expressed as an open reading frame of 1,287 bp, encoding a polypeptide of 429 amino acid residues with a predicted molecular mass of 50 kDa. It was expressed in Escherichia coli. BL21 and purified by metal affinity chromatography using a nickel-nitrilotriacetic acid column. Bioinformatics revealed that Ag85A-IL17A contained B cell epitopes of IL17A. Purified proteins Ag85A-IL17A and Ag85A were injected with adjuvants into mice then antibody responses in sera were measured by enzyme-linked immunosorbent assay and western blot assay. Experiments showed that purified protein Ag85-IL17A could bind to a standard IL17A antibody and could induce IL17A autoantibody in mice.
authors.affiliation: Department of Pediatrics, Affiliated 6th People’s Hospital, Shanghai Jiaotong University, 600 Yishan Road, Shanghai 200233, People’s Republic of China.
authors.forename: Rong
authors.initials: R
authors.lastname: Jin
authors.forename: Sheng
authors.initials: S
authors.lastname: Guo
authors.forename: Liangxia
authors.initials: L
authors.lastname: Wu
authors.forename: Xiaoyong
authors.initials: X
authors.lastname: Fan
authors.forename: Hui
authors.initials: H
authors.lastname: Ma
authors.forename: Douglas B
authors.initials: DB
authors.lastname: Lowrie
authors.forename: Jianhua
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000217:Acyltransferases / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000942:Antigens, Bacterial / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology*; D005260:Female; D020381:Interleukin-17 / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D001323:Autoantibodies; C508717:IL17A protein, human; D020381:Interleukin-17; D011994:Recombinant Proteins; D000217:Acyltransferases; C112580:antigen 85A, Mycobacterium tuberculosis