title: A novel immunoproteomics method for identifying in vivo-induced Campylobacter jejuni antigens using pre-adsorbed sera from infected patients.
abstract: BACKGROUND
  Campylobacter jejuni is an important food-borne and zoonotic pathogen with a worldwide distribution. Humans and chickens are hosts of this pathogen. At present, there is no ideal vaccine for controlling human campylobacteriosis or the carriage of C. jejuni by chickens. Bacterial in vivo-induced antigens are useful as potential vaccine candidates and biomarkers of virulence.
  
  
  METHODS
  In this study, we developed a novel systematic immunoproteomics approach to identify in vivo-induced antigens among the total cell proteins of C. jejuni using pre-adsorbed sera from patients infected with C. jejuni.
  
  
  RESULTS
  Overall, 14 immunoreactive spots were probed on a PVDF membrane using pre-adsorbed human sera against C. jejuni. Then, we excised these protein spots from a duplicate gel and identified using MALDI-TOF MS. In total, 14 in vivo-induced antigens were identified using PMF and BLAST analysis. The identified proteins include CadF (CadF-1 and CadF-2), CheW, TufB, DnaK, MetK, LpxB, HslU, DmsA, PorA, ProS, CJBH_0976, CSU_0396 and hypothetical protein cje135_05017. Real-time RT-PCR was performed on 9 genes to compare their expression levels in vivo and in vitro. The data showed that 8 of the 9 analyzed genes were significantly upregulated in vivo relative to in vitro.
  
  
  CONCLUSIONS
  We successfully developed a novel immunoproteomics method for identifying in vivo-induced Campylobacter jejuni antigens by using pre-adsorbed sera from infected patients.
  
  
  CONCLUSIONS
  This new analysis method may prove to be useful for identifying in vivo-induced antigens within any host infected by bacteria and will contribute to the development of new subunit vaccines.
authors.affiliation: Jiangsu Key Laboratory of Zoonosis, Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, 88 South Daxue Road, Yangzhou, Jiangsu 225009, PR China. Electronic address: huyuanqing1979@163.com.
authors.forename: Yuanqing
authors.initials: Y
authors.lastname: Hu
authors.forename: Yuwei
authors.initials: Y
authors.lastname: Shang
authors.forename: Jinlin
authors.initials: J
authors.lastname: Huang
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Fangzhe
authors.initials: F
authors.lastname: Ren
authors.forename: Yang
authors.initials: Y
authors.lastname: Jiao
authors.forename: Zhiming
authors.initials: Z
authors.lastname: Pan
authors.forename: Xin-An
authors.initials: XA
authors.lastname: Jiao
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002169:Campylobacter Infections / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D016123:Campylobacter jejuni / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D007106:Immune Sera / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015592:Immunoradiometric Assay / Q000379:methods*; D008875:Middle Aged; D015854:Up-Regulation
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D007106:Immune Sera
keywords: 2-DE; C. jejuni; CCDA; Campy blood-free selective medium; Campylobacter jejuni; DFI; DOC; GBS; Guillain–Barré syndrome; HRP; IEF; IVET; IVIAT; Immunoproteomics; In vivo-induced antigen; LOS; MALDI–TOF MS; MBP; NMRC; Naval Medical Research Center; PMF; STM; differential fluorescence induction; horseradish peroxidase; in vivo expression technology; in vivo-induced antigen technology; isoelectric focusing; lipooligosaccharides; maltose-binding protein; matrix-assisted laser desorption ionization–time of flight mass spectrometry; peptide mass fingerprinting; signature-tagged mutagenesis; sodium deoxycholate; two-dimensional echocardiography