To investigate the immunoprotective effects of the recombinant type A flagellin (FLA), the flaA gene of Legionella pneumophila serogroup 1 strain Paris was cloned into pET28a(+). Recombinant FLA (rFLA) was overexpressed in E. coli BL21 (DE3) and purified by Ni2+ exchange chromatography. Female BALB/c aged 6-8 weeks were immunized with 20 μg of rFLA. Nonimmunized mice along with mice inoculated with a sublethal dose of live L. pneumophila intravenously were considered as negative and positive controls, respectively. A significant serum antibody response was observed in female BALB/c mice immunized with rFLA. Production of IFN-γ and IL-12, and TNF-α in the serum and the splenocyte cultures, and antigen-specific splenocyte proliferation suggested a strong innate and adaptive cell-mediated immunity response in rFLA-immunized mice. Intravenous lethal challenge with L. pneumophila serogroup 1 (strain Paris) showed that 60% of mice immunized with rFLA survived in a 10-day follow-up survey. These results show that rFLA from L. pneumophila can elicit strong innate and adaptive immune responses and suggest the possibility of a long-term immunity against lethal challenge with L. pneumophila.

1.
Aghababa H, Mohabati Mobarez A, Khoramabadi N, Behmanesh M, Mahdavi M, Tebianian M, Nejati M: A comparative approach to strategies for cloning, expression, and purification of Mycobacterium tuberculosis mycolyl transferase 85B and evaluation of immune responses in BALB/c mice. Mol Biotechnol 2014;56:487-497.
[PubMed]
2.
Albert-Weissenberger C, Cazalet C, Buchrieser C: Legionella pneumophila - a human pathogen that co-evolved with fresh water protozoa. Cell Mol Life Sci 2007;64:432-448.
[PubMed]
3.
Blander SJ, Breiman RF, Horwitz MA: A live avirulent mutant Legionella pneumophila vaccine induces protective immunity against lethal aerosol challenge. J Clin Invest 1989;83:810-815.
[PubMed]
4.
Blander SJ, Horwitz MA: Vaccination with the major secretory protein of Legionella induces humoral and cell-mediated immune responses and protective immunity across different serogroups of Legionella pneumophila and different species of Legionella. J Immunol 1991;147:285-291.
[PubMed]
5.
Blander SJ, Szeto L, Shuman HA, Horwitz MA: An immunoprotective molecule, the major secretory protein of Legionella pneumophila, is not a virulence factor in a guinea pig model of Legionnaires' disease. J Clin Invest 1990;86:817-824.
[PubMed]
6.
Brieland JK, Engleberg NC, Huffnagle GB, Remick DG, Fantone JC: Host pathogen interactions in Legionnaires' disease: lessons learned from a murine animal model. Immunopharmacology 2000;48:249-252.
[PubMed]
7.
Brieland JK, Remick DG, LeGendre ML, Engleberg NC, Fantone JC: In vivo regulation of replicative Legionella pneumophila lung infection by endogenous interleukin-12. Infect Immun 1998;66:65-69.
[PubMed]
8.
Coscolla M, Fenollar J, Escribano I, Gonzalez-Candelas F: Legionellosis outbreak associated with asphalt paving machine, Spain, 2009. Emerg Infect Dis 2010;16:1381-1387.
[PubMed]
9.
Dietrich C, Heuner K, Brand BC, Hacker J, Steinert M: Flagellum of Legionella pneumophila positively affects the early phase of infection of eukaryotic host cells. Infect Immun 2001;69:2116-2122.
[PubMed]
10.
Fields BS, Benson RF, Besser RE: Legionella and Legionnaires' disease: 25 years of investigation. Clin Microbiol Rev 2002;15:506-526.
[PubMed]
11.
Fliermans CB, Cherry WB, Orrison LH, Smith SJ, Tison DL, Pope DH: Ecological distribution of Legionella pneumophila. Appl Environ Microbiol 1981;41:9-16.
[PubMed]
12.
Gomez-Valero L, Rusniok C, Buchrieser C: Legionella pneumophila: population genetics, phylogeny and genomics. Infect Genet Evol 2009;9:727-739.
[PubMed]
13.
Heath L, Chrisp C, Huffnagle G, LeGendre M, Osawa Y, Hurley M, Engleberg C, Fantone J, Brieland J: Effector mechanisms responsible for gamma interferon-mediated host resistance to Legionella pneumophila lung infection: the role of endogenous nitric oxide differs in susceptible and resistant murine hosts. Infect Immun 1996;64:5151-5160.
[PubMed]
14.
Heuner K, Bender-Beck L, Brand BC, Luck PC, Mann KH, Marre R, Ott M, Hacker J: Cloning and genetic characterization of the flagellum subunit gene (flaA) of Legionella pneumophila serogroup 1. Infect Immun 1995;63:2499-2507.
[PubMed]
15.
Heuner K, Steinert M, Marre R, Hacker J: Genomic structure and evolution of Legionella species. Curr Top Microbiol Immunol 2002;264:61-78.
[PubMed]
16.
Joller N, Sporri R, Hilbi H, Oxenius A: Induction and protective role of antibodies in Legionella pneumophila infection. Eur J Immunol 2007;37:3414-3423.
[PubMed]
17.
Kaufmann SH: Immunity to intracellular bacteria. Annu Rev Immunol 1993;11:129-163.
[PubMed]
18.
Klein TW, Newton C, Widen R, Friedman H: Delta 9-tetrahydrocannabinol injection induces cytokine-mediated mortality of mice infected with Legionella pneumophila. J Pharmacol Exp Ther 1993;267:635-640.
[PubMed]
19.
Merriam JJ, Mathur R, Maxfield-Boumil R, Isberg RR: Analysis of the Legionella pneumophilafliI gene: intracellular growth of a defined mutant defective for flagellum biosynthesis. Infect Immun 1997;65:2497-2501.
[PubMed]
20.
Newton C, McHugh S, Widen R, Nakachi N, Klein T, Friedman H: Induction of interleukin-4 (IL-4) by legionella pneumophila infection in BALB/c mice and regulation of tumor necrosis factor alpha, IL-6, and IL-1beta. Infect Immun 2000;68:5234-5240.
[PubMed]
21.
Newton CA, Klein TW, Friedman H: Secondary immunity to Legionella pneumophila and Th1 activity are suppressed by delta-9-tetrahydrocannabinol injection. Infect Immun 1994;62:4015-4020.
[PubMed]
22.
Ott M, Messner P, Heesemann J, Marre R, Hacker J: Temperature-dependent expression of flagella in Legionella. J Gen Microbiol 1991;137:1955-1961.
[PubMed]
23.
Pruckler JM, Benson RF, Moyenuddin M, Martin WT, Fields BS: Association of flagellum expression and intracellular growth of Legionella pneumophila. Infect Immun 1995;63:4928-4932.
[PubMed]
24.
Ricci ML, Torosantucci A, Scaturro M, Chiani P, Baldassarri L, Pastoris MC: Induction of protective immunity by Legionella pneumophila flagellum in an A/J mouse model. Vaccine 2005;23:4811-4820.
[PubMed]
25.
Skerrett SJ, Bagby GJ, Schmidt RA, Nelson S: Antibody-mediated depletion of tumor necrosis factor-alpha impairs pulmonary host defenses to Legionella pneumophila. J Infect Dis 1997;176:1019-1028.
[PubMed]
26.
Skerrett SJ, Martin TR: Roles for tumor necrosis factor alpha and nitric oxide in resistance of rat alveolar macrophages to Legionella pneumophila. Infect Immun 1996;64:3236-3243.
[PubMed]
27.
Susa M, Ticac B, Rukavina T, Doric M, Marre R: Legionella pneumophila infection in intratracheally inoculated T cell-depleted or -nondepleted A/J mice. J Immunol 1998;160:316-321.
[PubMed]
28.
Xu JN, Yang ZW, Chen JP, Chen DL, Wang T, Liu MJ, Zhang L, Yang BB: Protective immunity against Legionnaires' disease in an A/J mouse model using a DNA vaccine composed of an outer membrane protein (29 kDa) and the pilE fusion protein. Diagn Microbiol Infect Dis 2012;73:9-15.
[PubMed]
29.
Xu Y, Guan W, Xu JN, Cao DP, Yang BB, Chen DL, Chen JP: Evaluation of the protective immunity of the Legionella pneumophila recombinant protein FlaA/MompS/PilE in an A/J mouse model. Vaccine 2011;29:4051-4057.
[PubMed]
30.
Yang ZW, Chen JP, Wang T, Tian Y: Immune response and protective efficacy in BALB/c mice induced by DNA vaccine encoding immunogenic protein of Legionella pneumophila. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2007;23:209-212.
[PubMed]
31.
Zhang L, Chen JP, Zhang L, Wang T, Liu MJ, Tian Y: Cloning and prokaryotic expression of flagellum subunit gene (flaA) of Legionella pneumophila. Sichuan Da Xue Xue Bao Yi Xue Ban 2007;38:194-197.
[PubMed]
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