Identification, Characterization and Immunogenicity of an O-Antigen Capsular Polysaccharide of Francisella tularensis

被引:72
作者
Apicella, Michael A. [1 ]
Post, Deborah M. B. [2 ]
Fowler, Andrew C. [1 ]
Jones, Bradley D. [1 ]
Rasmussen, Jed A. [1 ]
Hunt, Jason R. [1 ]
Imagawa, Sayaka [3 ]
Choudhury, Biswa [3 ]
Inzana, Thomas J. [4 ]
Maier, Tamara M. [5 ]
Frank, Dara W. [5 ]
Zahrt, Thomas C. [5 ]
Chaloner, Kathryn [6 ]
Jennings, Michael P. [7 ]
McLendon, Molly K. [1 ]
Gibson, Bradford W. [2 ]
机构
[1] Univ Iowa, Dept Microbiol, Iowa City, IA 52242 USA
[2] Buck Inst Age Res, Novato, CA USA
[3] Univ Calif San Diego, San Diego, CA 92103 USA
[4] Virginia Polytech Inst & State Univ, Virginia Maryland Reg Coll Vet Med, Blacksburg, VA 24061 USA
[5] Med Coll Wisconsin, Ctr Biopreparedness & Infect Dis, Milwaukee, WI 53226 USA
[6] Univ Iowa, Coll Publ Hlth, Dept Biostat, Iowa City, IA 52242 USA
[7] Griffith Univ, Inst Glyc, Gold Coast, Australia
来源
PLOS ONE | 2010年 / 5卷 / 07期
关键词
STRUCTURAL-ANALYSIS; TULAREMIA VACCINE; ESCHERICHIA-COLI; DEFICIENT MUTANT; LIPID-A; LVS; BACTERIUM; VIRULENCE; BIOSYNTHESIS; DISEASE;
D O I
10.1371/journal.pone.0011060
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Capsular polysaccharides are important factors in bacterial pathogenesis and have been the target of a number of successful vaccines. Francisella tularensis has been considered to express a capsular antigen but none has been isolated or characterized. We have developed a monoclonal antibody, 11B7, which recognizes the capsular polysaccharide of F. tularensis migrating on Western blot as a diffuse band between 100 kDa and 250 kDa. The capsule stains poorly on SDS-PAGE with silver stain but can be visualized using ProQ Emerald glycoprotein stain. The capsule appears to be highly conserved among strains of F. tularensis as antibody 11B7 bound to the capsule of 14 of 14 F. tularensis type A and B strains on Western blot. The capsular material can be isolated essentially free of LPS, is phenol and proteinase K resistant, ethanol precipitable and does not dissociate in sodium dodecyl sulfate. Immunoelectron microscopy with colloidal gold demonstrates 11B7 circumferentially staining the surface of F. tularensis which is typical of a polysaccharide capsule. Mass spectrometry, compositional analysis and NMR indicate that the capsule is composed of a polymer of the tetrasaccharide repeat, 4)-alpha-D-GalNAcAN-(1->4)-alpha-D-GalNAcAN-(1->3)-beta-D-QuiNAc-(1->2)-beta-D-Qui4NFm-(1-, which is identical to the previously described F. tularensis O-antigen subunit. This indicates that the F. tularensis capsule can be classified as an O-antigen capsular polysaccharide. Our studies indicate that F. tularensis O-antigen glycosyltransferase mutants do not make a capsule. An F. tularensis acyltransferase and an O-antigen polymerase mutant had no evidence of an O-antigen but expressed a capsular antigen. Passive immunization of BALB/c mice with 75 mu g of 11B7 protected against a 150 fold lethal challenge of F. tularensis LVS. Active immunization of BALB/c mice with 10 mu g of capsule showed a similar level of protection. These studies demonstrate that F. tularensis produces an O-antigen capsule that may be the basis of a future vaccine.
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页数:18
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