Francisella tularensis subspecies tularensis is a highly virulent intracellular bacterial pathogen, causing the disease tularemia. However, a safe and effective vaccine for routine application against F. tularensis has not yet been developed. We have recently constructed the deletion mutants for the DsbA homolog protein (ΔdsbA/FSC200) and a hypothetical protein IglH (ΔiglH/FSC200) in the type B F. tularensis subsp. holarctica FSC200 strain, which exerted different protection capacity against parental virulent strain. In this study, we further investigated the immunological correlates for these different levels of protection provided by ΔdsbA/FSC200 and ΔiglH/FSC200 mutants. Our results show that ΔdsbA/FSC200 mutant, but not ΔiglH/FSC200 mutant, induces an early innate inflammatory response leading to strong Th1-like antibody response. Furthermore, vaccination with ΔdsbA/FSC200 mutant, but not with ΔiglH/FSC200, elicited protection against the subsequent challenge with type A SCHU S4 strain in mice. An immunoproteomic approach was used to map a spectrum of antigens targeted by Th1-like specific antibodies, and more than 80 bacterial antigens, including novel ones, were identified. Comparison of tularemic antigens recognized by the ΔdsbA/FSC200 post-vaccination and the SCHU S4 post-challenge sera then revealed the existence of 22 novel SCHU S4 specific antibody clones.

Bacteriology Division U S Army Medical Research Institute of Infectious Diseases Fort Detrick MD 21702 5011 USA

Department of Molecular Pathology Faculty of Military Health Sciences University of Defence Hradec Kralove Trebesska 1575 Hradec Kralove 500 01 Czech Republic

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Akimana C, Al-Khodor S, Abu Kwaik Y. Host factors required for modulation of phagosome biogenesis and proliferation of Francisella tularensis within the cytosol. PLoS One. 2010;5:e11025. PubMed PMC

Balonova L, Hernychova L, Mann BF, et al. Multimethodological approach to identification of glycoproteins from the proteome of Francisella tularensis, an intracellular microorganism. J Proteome Res. 2010;9:1995–2005. PubMed PMC

Broms JE, Meyer L, Sun K, et al. Unique substrates secreted by the type VI secretion system of Francisella tularensis during intramacrophage infection. PLoS One. 2012;7:e50473. PubMed PMC

Chong A, Child R, Wehrly TD, et al. Structure-function analysis of DipA, a virulence factor required for intracellular replication. PLoS One. 2013;8:e67965. PubMed PMC

Dennis DT, Inglesby TV, Henderson DA, et al. Tularemia as a biological weapon: medical and public health management. JAMA. 2001;285:2763–73. PubMed

Elkins KL, Cooper A, Colombini SM, et al. In vivo clearance of an intracellular bacterium, Francisella tularensis LVS, is dependent on the p40 subunit of interleukin-12 (IL-12) but not on IL-12 p70. Infect Immun. 2002;70:1936–48. PubMed PMC

Evans ME, Gregory DW, Schaffner W, et al. Tularemia: a 30-year experience with 88 cases. Medicine. 1985;64:251–69. PubMed

Forslund AL, Salomonsson EN, Golovliov I, et al. The type IV pilin, PilA, is required for full virulence of Francisella tularensis subspecies tularensis. BMC Microbiol. 2010;10:227. PubMed PMC

Fulton KM, Zhao X, Petit MD, et al. Immunoproteomic analysis of the human antibody response to natural tularemia infection with Type A or Type B strains or LVS vaccination. Int J Med Microbiol. 2011;301:591–601. PubMed PMC

Furuya Y, Kirimanjeswara GS, Roberts S, et al. Increased susceptibility of IgA-deficient mice to pulmonary Francisella tularensis live vaccine strain infection. Infect Immun. 2013;81:3434–41. PubMed PMC

Golovliov I, Twine SM, Shen H, et al. A deltaclpB mutant of Francisella tularensis subspecies holarctica strain, FSC200, is a more effective live vaccine than F. tularensis LVS in a mouse respiratory challenge model of tularemia. PLoS One. 2013;8:e78671. PubMed PMC

Hubalek M, Hernychova L, Brychta M, et al. Comparative proteome analysis of cellular proteins extracted from highly virulent Francisella tularensis ssp. tularensis and less virulent F. tularensis ssp. holarctica and F. tularensis ssp. mediaasiatica. Proteomics. 2004;4:3048–60. PubMed

Johansson A, Berglund L, Sjostedt A, et al. Ciprofloxacin for treatment of tularemia. Clin Infect Dis. 2001;33:267–8. PubMed

Kilmury SL, Twine SM. The Francisella tularensis proteome and its recognition by antibodies. Front Microbiol. 2010;1:143. PubMed PMC

Konecna K, Hernychova L, Reichelova M, et al. Comparative proteomic profiling of culture filtrate proteins of less and highly virulent Francisella tularensis strains. Proteomics. 2010;10:4501–11. PubMed

Marohn ME, Barry EM. Live attenuated tularemia vaccines: recent developments and future goals. Vaccine. 2013;31:3485–91. PubMed PMC

Melillo AA, Mahawar M, Sellati TJ, et al. Identification of Francisella tularensis live vaccine strain CuZn superoxide dismutase as critical for resistance to extracellularly generated reactive oxygen species. J Bacteriol. 2009;191:6447–56. PubMed PMC

Mohapatra NP, Soni S, Rajaram MV, et al. Type A Francisella tularensis acid phosphatases contribute to pathogenesis. PLoS One. 2013;8:e56834. PubMed PMC

Oyston PC, Sjostedt A, Titball RW. Tularaemia: bioterrorism defence renews interest in Francisella tularensis. Nat Rev Microbiol. 2004;2:967–78. PubMed

Pavkova I, Reichelova M, Larsson P, et al. Comparative proteome analysis of fractions enriched for membrane-associated proteins from Francisella tularensis subsp. tularensis and F. tularensis subsp. holarctica strains. J Proteome Res. 2006;5:3125–34. PubMed

Pechous RD, McCarthy TR, Zahrt TC. Working toward the future: insights into Francisella tularensis pathogenesis and vaccine development. Microbiol Mol Biol R. 2009;73:684–711. PubMed PMC

Pelletier N, Raoult D, La Scola B. Specific recognition of the major capsid protein of Acanthamoeba polyphaga mimivirus by sera of patients infected by Francisella tularensis. FEMS Microbiol Lett. 2009;297:117–23. PubMed

Qin A, Scott DW, Thompson JA, et al. Identification of an essential Francisella tularensis subsp. tularensis virulence factor. Infect Immun. 2009;77:152–61. PubMed PMC

Robertson GT, Child R, Ingle C, et al. IglE Is an outer membrane-associated lipoprotein essential for intracellular survival and murine virulence of type A Francisella tularensis. Infect Immun. 2013;81:4026–40. PubMed PMC

Rodriguez AR, Yu JJ, Guentzel MN, et al. Mast cell TLR2 signaling is crucial for effective killing of Francisella tularensis. J Immunol. 2012;188:5604–11. PubMed PMC

Russell P, Eley SM, Fulop MJ, et al. The efficacy of ciprofloxacin and doxycycline against experimental tularaemia. J Antimicrob Chemoth. 1998;41:461–5. PubMed

Ryden P, Twine S, Shen H, et al. Correlates of protection following vaccination of mice with gene deletion mutants of Francisella tularensis subspecies tularensis strain, SCHU S4 that elicit varying degrees of immunity to systemic and respiratory challenge with wild-type bacteria. Mol Immunol. 2013;54:58–67. PubMed

Schmidt M, Klimentova J, Rehulka P, et al. Francisella tularensis subsp. holarctica DsbA homologue: a thioredoxin-like protein with chaperone function. Microbiology. 2013;159:2364–74. PubMed

Shimizu T, Kida Y, Kuwano K. A dipalmitoylated lipoprotein from Mycoplasma pneumoniae activates NF-kappa B through TLR1, TLR2, and TLR6. J Immunol. 2005;175:4641–6. PubMed

Skyberg JA, Rollins MF, Samuel JW, et al. Interleukin-17 protects against the Francisella tularensis live vaccine strain but not against a virulent F. tularensis type A strain. Infect Immun. 2013;81:3099–105. PubMed PMC

Straskova A, Cerveny L, Spidlova P, et al. Deletion of IglH in virulent Francisella tularensis subsp. holarctica FSC200 strain results in attenuation and provides protection against the challenge with the parental strain. Microbes Infect. 2012;14:177–87. PubMed

Straskova A, Pavkova I, Link M, et al. Proteome analysis of an attenuated Francisella tularensis dsbA mutant: identification of potential DsbA substrate proteins. J Proteome Res. 2009;8:5336–46. PubMed

Tarnvik A, Berglund L. Tularaemia. Eur Respir J. 2003;21:361–73. PubMed

Thathiah P, Sanapala S, Rodriguez AR, et al. Non-FcepsilonR bearing mast cells secrete sufficient interleukin-4 to control Francisella tularensis replication within macrophages. Cytokine. 2011;55:211–20. PubMed PMC

Weiss DS, Brotcke A, Henry T, et al. In vivo negative selection screen identifies genes required for Francisella virulence. P Natl Acad Sci USA. 2007;104:6037–42. PubMed PMC

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Tularemia vaccines