Abstract
MglA (Macrophage Growth Locus A) is a pleiotropic transcription factor controlling the expression pattern of more than 100 genes in Francisella novicida, including all the ORFs located on the pathogenicity island. To further probe the role of MglA in the pathogenicity of the related strain Francisella tularensis LVS, we generated a mutant LVS strain in which the mglA locus was disrupted by insertion of a non-polar selectable marker cassette.
In vitro and in vivo analysis of the phenotype associated with the mglA null mutation in comparison to the wild-type parental strain and to a complementation strain (ΔmglA:mglA), established that the mutated strain ΔmglA: (i) cannot multiply in vitro in macrophages, (ii) is severely attenuated in a murine model of infection, exhibiting over 10,000 and 10,000,000 fold decrease in virulence by intranasal (IN) administration, and intraperitoneal (IP) route of infection respectively, (iii) unlike wild type LVS, the mutant strain cannot multiply in the lungs, liver and spleen of infected animals following IP administration, (iv) the ΔmglA mutant do not disseminate to target organs (e.g. liver and spleen) following IN administration, (v) Infection by ΔmglA bacteria elicits a significant humoral response, and systemic IP administration of high doses of ΔmglA cells results in full protection of animals against a subsequent IP challenge with high doses of the virulent wild-type strain.
These results indicate that MglA plays a major role in F. tularensis LVS virulence as previously shown for F. novicida. Our studies suggest that inactivation of MglA may serve as a platform for the development of an improved attenuated vaccine of virulent F. tularensis strains.
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References
Baron, G. S. & Nano, F. E. (1998) MglA and MglB are required for the intramacrophage growth of Francisella novicida. Mol Microbiol, 29, 247–59.
Ben Nasr, A., Haithcoat, J., Masterson, J. E., Gunn, J. S., Eaves-Pyles, T. & Klimpel, G. R. (2006) Critical role for serum opsonins and complement receptors CR3 (CD11b/CD18) and CR4 (CD11c/CD18) in phagocytosis of Francisella tularensis by human dendritic cells (DC): uptake of Francisella leads to activation of immature DC and intracellular survival of the bacteria. J Leukoc Biol, 80, 774–86.
Bonquist, L., Lindgren, H., Golovliov, I., Guina, T. & Sjöstedt, A. (2008) MglA and Igl proteins contribute to the modulation of Francisella tularensis live vaccine strain-containing phagosomes in murine macrophages. Infect Immun, 76, 3502–10.
Brotcke, A., Weiss, D. S., Kim, C. C., Chain, P., Malfatti, S., Garcia, E. & Monack, D. M. (2006) Identification of MglA-regulated genes reveals novel virulence factors in Francisella tularensis. Infect Immun, 74, 6642–55.
Buddingh, G. J. & Womack, F. C. (1941) Observations on the infection of chick embryos with Bacterium Tularense, Brucella, and Pasteurella Pestis. J Exp Med, 74, 213–222.
Charity, J. C., Costante-Hamm, M. M., Balon, E. L., Boyd, D. H., Rubin, E. J. & Dove, S. L. (2007) Twin RNA polymerase-associated proteins control virulence gene expression in Francisella tularensis. PLoS Pathog, 3, e84.
Conlan, J. W. & Oyston, P. C. (2007) Vaccines against Francisella tularensis. Ann NY Acad Sci, 1105, 325–50.
Councilman, W. T. S. & Strong, R. P (1921) Plague-like infections in rodents. Trans Assoc Am Phys., 36, 135–43.
Ellis, J., Oyston, P. C., Green, M. & Titball, R. W. (2002) Tularemia. Clin Microbiol Rev, 15, 631–46.
Forsman, M., Sandstrom, G. & Sjöstedt, A. (1994) Analysis of 16S ribosomal DNA sequences of Francisella strains and utilization for determination of the phylogeny of the genus and for identification of strains by PCR. Int J Syst Bacteriol, 44, 38–46.
Fortier, A. H., Green, S. J., Polsinelli, T., Jones, T. R., Crawford, R. M., Leiby, D. A., Elkins, K. L., Meltzer, M. S. & Nacy, C. A. (1994) Life and death of an intracellular pathogen: Francisella tularensis and the macrophage. Immunol Ser, 60, 349–61.
Francis, E. E. (1927) Microscopic changes of tularemia in the tick, Dermacentor andersoni and the bedbug, Cimex lectularius. Pub Health Rep, 42, 2763–72.
Guina, T., Radulovic, D., Bahrami, A. J., Bolton, D. L., Rohmer, L., Jones-Isaac, K. A., Chen, J., Gallagher, L. A., Gallis, B., Ryu, S., Taylor, G. K., Brittnacher, M. J., Manoil, C. & Goodlett, D. R. (2007) MglA regulates Francisella tularensis subsp. novicida (Francisella novicida) response to starvation and oxidative stress. J Bacteriol, 189, 6580–6.
Larsson, P., Oyston, P. C., Chain, P., Chu, M. C., Duffield, M., Fuxelius, H. H., Garcia, E., Halltorp, G., Johansson, D., Isherwood, K. E., Karp, P. D., Larsson, E., Liu, Y., Michell, S., Prior, J., Prior, R., Malfatti, S., Sjöstedt, A., Svensson, K., Thompson, N., Vergez, L., Wagg, J. K., Wren, B. W., Lindler, L. E., Andersson, S. G., Forsman, M. & Titball, R. W. (2005) The complete genome sequence of Francisella tularensis, the causative agent of tularemia. Nat Genet, 37, 153–9.
Lauriano, C. M., Barker, J. R., Yoon, S. S., Nano, F. E., Arulanandam, B. P., Hassett, D. J. & Klose, K. E. (2004) MglA regulates transcription of virulence factors necessary for Francisella tularensis intraamoebae and intramacrophage survival. Proc Natl Acad Sci USA, 101, 4246–9.
Lindemann, S. R., Mclendon, M. K., Apicella, M. A. & Jones, B. D. (2007) An in vitro model system used to study adherence and invasion of Francisella tularensis live vaccine strain in nonphagocytic cells. Infect Immun, 75, 3178–82.
Mariathasan, S., Weiss, D. S., Dixit, V. M. & Monack, D. M. (2005) Innate immunity against Francisella tularensis is dependent on the ASC/caspase-1 axis. J Exp Med, 202, 1043–9.
Nano, F. E., Zhang, N., Cowley, S. C., Klose, K. E., Cheung, K. K., Roberts, M. J., Ludu, J. S., Letendre, G. W., Meierovics, A. I., Stephens, G. & Elkins, K. L. (2004) A Francisella tularensis pathogenicity island required for intramacrophage growth. J Bacteriol, 186, 6430–6.
Oyston, P. C., Sjöstedt, A. & Titball, R. W. (2004) Tularaemia: bioterrorism defence renews interest in Francisella tularensis. Nat Rev Microbiol, 2, 967–78.
Ritter, D. B. G. & Gerloff, R. K. (1966) Deoxyribonucleic acid hybridization among some species of the genus Pasteruella. J Bacteriol, 92, 1838–9.
Sandstrom, G., Sjöstedt, A., Johansson, T., Kuoppa, K. & Williams, J. C. (1992) Immunogenicity and toxicity of lipopolysaccharide from Francisella tularensis LVS. FEMS Microbiol Immunol 5, 201–10.
Santic, M., Molmeret, M., Klose, K. E., Jones, S. & Kwaik, Y. A. (2005) The Francisella tularensis pathogenicity island protein IglC and its regulator MglA are essential for modulating phagosome biogenesis and subsequent bacterial escape into the cytoplasm. Cell Microbiol, 7, 969–79.
Shepard, C. C. (1959) Nonacid-fast bacteria and HeLa cells: their uptake and subsequent intracellular growth. J Bacteriol, 77, 701–14.
Tigertt, W. D. (1962) Soviet viable Pasteurella tularensis vaccines. Bacterio Rev, 26, 354–73.
West, T. E., Pelletier, M. R., Majure, M. C., Lembo, A., Hajjar, A. M. & Skerrett, S. J. (2008) Inhalation of Francisella novicida Delta mglA causes replicative infection that elicits innate and adaptive responses but is not protective against invasive pneumonic tularemia. Microbes Infect, 10, 773–80.
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Cohen, O. et al. (2010). Effect of Disruption of mglA on the Virulence and Immunogenicity of the Francisella tularensis Live Vaccine Strain (LVS). In: Shafferman, A., Ordentlich, A., Velan, B. (eds) The Challenge of Highly Pathogenic Microorganisms. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9054-6_24
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DOI: https://doi.org/10.1007/978-90-481-9054-6_24
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