title: Proteomic analysis of anti-Francisella tularensis LVS antibody response in murine model of tularemia.
abstract: Francisella tularensis live vaccine strain infection of mice has been established as an experimental model of tularemia that is suitable for studies of immune mechanisms against the intracellular pathogen. In this study, the model was used to explore immunogenic repertoire of F. tularensis with the aim of identifying new molecules able to activate the host immune system, potential bacterial markers with vaccine, and diagnostic applications. Immunoproteomic approach based on the combination of two-dimensional gel electrophoresis, immunoblotting, and mass spectrometry was applied. Globally, 36 different proteins were identified, which strongly reacted with sera from experimentally infected mice, including several putative virulence markers of intracellular pathogens as nucleoside diphosphate kinase, isocitrate dehydrogenase, RNA-binding protein Hfq, and molecular chaperone ClpB. Of them, 27 proteins are described for the first time as immunorelevant Francisella proteins. When comparing murine immunoproteome of F. tularensis with our previous data from human patients, 25 of the total of 50 identified murine sera immunoreactive spots were recognized by human sera collected from patients suffering from tularemia, as well. Immune sera from two Lps gene congenic strains of mice, C3H/HeN (Lpsn) and C3H/HeJ (Lpsd), represented murine immunoproteome in this study. The spectrum of immunoreactive spots detected by two-dimensional immunoblotting varied throughout the course of infection depending on murine strain. Nevertheless, the antibody patterns of the two strains showed significant homogeneity in being directed against almost identical subset of antigens.
authors.affiliation: Proteome Center for the Study of Intracellular Parasitism of Bacteria, Purkyne Military Medical Academy, Hradec Králové, Czech Republic.
authors.forename: Jana
authors.initials: J
authors.lastname: Havlasová
authors.forename: Lenka
authors.initials: L
authors.lastname: Hernychová
authors.forename: Martin
authors.initials: M
authors.lastname: Brychta
authors.forename: Martin
authors.initials: M
authors.lastname: Hubálek
authors.forename: Jurai
authors.initials: J
authors.lastname: Lenco
authors.forename: Pär
authors.initials: P
authors.lastname: Larsson
authors.forename: Margaretha
authors.initials: M
authors.lastname: Lundqvist
authors.forename: Mats
authors.initials: M
authors.lastname: Forsman
authors.forename: Zulana
authors.initials: Z
authors.lastname: Krocová
authors.forename: Jiri
authors.initials: J
authors.lastname: Stulík
authors.forename: Aks
authors.initials: A
authors.lastname: Macela
mesh_terms: D000818:Animals; D020296:Animals, Congenic; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D019295:Computational Biology; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005604:Francisella tularensis / Q000276:immunology*; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D013058:Mass Spectrometry; D051379:Mice; D008809:Mice, Inbred C3H; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014406:Tularemia / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007106:Immune Sera