title: Immunodominant Francisella tularensis antigens identified using proteome microarray.
abstract: Stimulation of protective immune responses against intracellular pathogens is difficult to achieve using non-replicating vaccines. BALB/c mice immunized by intramuscular injection with killed Francisella tularensis (live vaccine strain) adjuvanted with preformed immune stimulating complexes admixed with CpG, were protected when systemically challenged with a highly virulent strain of F. tularensis (Schu S4). Serum from immunized mice was used to probe a whole proteome microarray in order to identify immunodominant antigens. Eleven out of the top 12 immunodominant antigens have been previously described as immunoreactive in F. tularensis. However, 31 previously unreported immunoreactive antigens were revealed using this approach. Twenty four (50%) of the ORFs on the immunodominant hit list belonged to the category of surface or membrane associated proteins compared to only 22% of the entire proteome. There were eight hypothetical protein hits and eight hits from proteins associated with different aspects of metabolism. The chip also allowed us to readily determine the IgG subclass bias, towards individual or multiple antigens, in protected and unprotected animals. These data give insight into the protective immune response and have potentially important implications for the rational design of non-living vaccines for tularemia and other intracellular pathogens.
authors.affiliation: Defence Science and Technology Laboratory, Porton Down, Salisbury, UK. jeeyles@dstl.gov.uk
authors.forename: Jim E
authors.initials: JE
authors.lastname: Eyles
authors.forename: Berkay
authors.initials: B
authors.lastname: Unal
authors.forename: Margaret Gill
authors.initials: MG
authors.lastname: Hartley
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Newstead
authors.forename: Helen
authors.initials: H
authors.lastname: Flick-Smith
authors.forename: Joann L
authors.initials: JL
authors.lastname: Prior
authors.forename: Petra C F
authors.initials: PC
authors.lastname: Oyston
authors.forename: Arlo
authors.initials: A
authors.lastname: Randall
authors.forename: Yunxiang
authors.initials: Y
authors.lastname: Mu
authors.forename: Siddiqua
authors.initials: S
authors.lastname: Hirst
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Molina
authors.forename: D Huw
authors.initials: DH
authors.lastname: Davies
authors.forename: Tim
authors.initials: T
authors.lastname: Milne
authors.forename: Kate F
authors.initials: KF
authors.lastname: Griffin
authors.forename: Pierre
authors.initials: P
authors.lastname: Baldi
authors.forename: Rick W
authors.initials: RW
authors.lastname: Titball
authors.forename: Philip L
authors.initials: PL
authors.lastname: Felgner
mesh_terms: D000276:Adjuvants, Immunologic; D000536:Aluminum Hydroxide / Q000276:immunology; D000818:Animals; D001426:Bacterial Proteins / Q000276:immunology / Q000378:metabolism; D005260:Female; D005604:Francisella tularensis / Q000276:immunology* / Q000378:metabolism; D016882:ISCOMs / Q000276:immunology; D016056:Immunodominant Epitopes / Q000032:analysis* / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007371:Interferon-gamma / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009838:Oligodeoxyribonucleotides / Q000276:immunology; D040081:Protein Array Analysis / Q000379:methods*; D020543:Proteome / Q000276:immunology / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D013154:Spleen / Q000166:cytology / Q000276:immunology; D016019:Survival Analysis; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D014406:Tularemia / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control; D014611:Vaccination
chemical_list: D000276:Adjuvants, Immunologic; D001426:Bacterial Proteins; D016882:ISCOMs; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G; D007376:Interleukin-2; D009838:Oligodeoxyribonucleotides; C483020:ProMune; D020543:Proteome; D015847:Interleukin-4; D000536:Aluminum Hydroxide; D007371:Interferon-gamma