title: Anthrax LFn-PA Hybrid Antigens: Biochemistry, Immunogenicity, and Protection Against Lethal Ames Spore Challenge in Rabbits.
abstract: We describe a novel hybrid anthrax toxin approach that incorporates multiple components into a single vaccine product. The key domains of protective antigen (PA) and lethal factor (LF) that may be critical for inducing protective immunity are combined into one recombinant molecule. Two LF N-terminal domain-PA hybrids, one with wild-type PA and another with furin cleavage-minus PA, were expressed in E. coli and purified in a native form. Both the hybrids bind to the extracellular domain of the host receptor, CMG2; the wild-type hybrid can be cleaved by furin exposing the LF interacting domain, allowing it to oligomerize into lethal toxin as well as translocation pore-like complexes. The hybrid antigens are immunogenic in Dutch-belted rabbits, eliciting strong PA-specific and LF-specific antibodies. However, the lethal toxin neutralizing antibody titers are 3-7 times lower than those elicited by PA-alum. The hybrid antigens conferred 100% (6/6) protection in rabbits challenged intranasally with a 100 LD(50) dose of Bacillus anthracis Ames strain spores.
authors.affiliation: Department of Biology, 103 McCort Ward Hall, The Catholic University of America, 620 Michigan Ave., NE, Washington, DC 20064, USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Li
authors.forename: Kristina K
authors.initials: KK
authors.lastname: Peachman
authors.forename: Laurie
authors.initials: L
authors.lastname: Sower
authors.forename: Stephen H
authors.initials: SH
authors.lastname: Leppla
authors.forename: Sathish B
authors.initials: SB
authors.lastname: Shivachandra
authors.forename: Gary R
authors.initials: GR
authors.lastname: Matyas
authors.forename: Johnny W
authors.initials: JW
authors.lastname: Peterson
authors.forename: Carl R
authors.initials: CR
authors.lastname: Alving
authors.forename: Mangala
authors.initials: M
authors.lastname: Rao
authors.forename: Venigalla B
authors.initials: VB
authors.lastname: Rao
references.citation: Protein Expr Purif. 2000 Apr;18(3):293-302
references.pmid: 10733882
references.citation: Arch Intern Med. 1998 Mar 9;158(5):429-34
references.pmid: 9508220
references.citation: Clin Vaccine Immunol. 2008 Apr;15(4):612-6
references.pmid: 18256208
references.citation: J Ind Microbiol Biotechnol. 2002 Apr;28(4):232-8
references.pmid: 11986925
references.citation: Gene. 1988 Sep 30;69(2):287-300
references.pmid: 3148491
references.citation: Infect Immun. 2007 Nov;75(11):5425-33
references.pmid: 17646360
references.citation: J Biol Chem. 2001 Mar 16;276(11):8371-6
references.pmid: 11113126
references.citation: Cell Microbiol. 2007 Apr;9(4):977-87
references.pmid: 17381430
references.citation: Vaccine. 2002 Jan 31;20(9-10):1412-20
references.pmid: 11818160
references.citation: Methods Enzymol. 1988;165:103-16
references.pmid: 3148094
references.citation: Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10277-81
references.pmid: 1438214
references.citation: Nature. 2004 Aug 19;430(7002):905-8
references.pmid: 15243628
references.citation: J Biol Chem. 2005 Dec 2;280(48):39897-906
references.pmid: 16188881
references.citation: J Appl Microbiol. 1999 Aug;87(2):304
references.pmid: 10475975
references.citation: Cell Mol Life Sci. 2006 Oct;63(19-20):2237-48
references.pmid: 16964578
references.citation: Infect Immun. 1986 May;52(2):356-63
references.pmid: 3084381
references.citation: J Biol Chem. 2006 Jan 20;281(3):1630-5
references.pmid: 16293620
references.citation: Vaccine. 2004 Apr 16;22(13-14):1604-8
references.pmid: 15068841
references.citation: Infect Immun. 2006 Jan;74(1):794-7
references.pmid: 16369043
references.citation: Proc Natl Acad Sci U S A. 2005 Nov 8;102(45):16409-14
references.pmid: 16251269
references.citation: Infect Immun. 2001 Jul;69(7):4509-15
references.pmid: 11401993
references.citation: Annu Rev Cell Dev Biol. 2003;19:45-70
references.pmid: 14570563
references.citation: Virology. 2007 Oct 25;367(2):422-7
references.pmid: 17624389
references.citation: Annu Rev Biochem. 2007;76:243-65
references.pmid: 17335404
references.citation: Expert Rev Mol Med. 2006 Apr 11;8(7):1-18
references.pmid: 16608555
references.citation: Microbiol Mol Biol Rev. 2004 Dec;68(4):617-29
references.pmid: 15590776
references.citation: Infect Immun. 2004 Feb;72(2):1181-3
references.pmid: 14742571
references.citation: Med Microbiol Immunol. 1988;177(5):293-303
references.pmid: 3139974
references.citation: Appl Microbiol. 1963 Jul;11:330-4
references.pmid: 13972634
references.citation: Gene. 1989 Apr 15;77(1):61-8
references.pmid: 2744488
references.citation: Gene. 1986;44(1):71-8
references.pmid: 3021591
references.citation: Nature. 2001 Nov 8;414(6860):225-9
references.pmid: 11700562
references.citation: J Biol Chem. 2001 May 25;276(21):17976-84
references.pmid: 11278833
references.citation: Hum Antibodies. 2003;12(4):129-35
references.pmid: 15156101
references.citation: Vaccine. 1991 Aug;9(8):533-9
references.pmid: 1771966
references.citation: Protein Sci. 2006 Sep;15(9):2190-200
references.pmid: 16943448
references.citation: Infect Immun. 2006 Feb;74(2):1016-24
references.pmid: 16428748
references.citation: J Bacteriol. 2001 Mar;183(6):2111-6
references.pmid: 11222612
references.citation: Biochemistry. 1999 Aug 10;38(32):10432-41
references.pmid: 10441138
references.citation: Curr Opin Microbiol. 2004 Feb;7(1):19-24
references.pmid: 15036135
references.citation: Biologicals. 2004 Mar;32(1):17-27
references.pmid: 15026022
references.citation: Vaccine. 2007 Jan 26;25(7):1225-35
references.pmid: 17069938
references.citation: Cell Microbiol. 2000 Jun;2(3):251-8
references.pmid: 11207581
references.citation: Infect Immun. 2005 Sep;73(9):5978-87
references.pmid: 16113318
references.citation: Proc Natl Acad Sci U S A. 2003 Apr 29;100(9):5170-4
references.pmid: 12700348
references.citation: Br J Exp Pathol. 1954 Apr;35(2):144-52
references.pmid: 13149735
references.citation: J Biol Chem. 2000 Nov 24;275(47):36741-9
references.pmid: 10958789