Recombinant Bacillus subtilis expressing the Clostridium perfringens alpha toxoid is a candidate orally delivered vaccine against necrotic enteritis.
Recombinant Bacillus subtilis endospores have been used to vaccinate against tetanus and anthrax. In this work, we have developed spores that could be used to vaccinate against Clostridium perfringens alpha toxin and that could be used to protect against gas gangrene in humans and necrotic enteritis in poultry. The primary active agent in both cases is alpha toxin. A carboxy-terminal segment of the alpha toxin gene (cpa) fused to the glutathione-S-transferase (GST) gene was cloned in B. subtilis such that the encoded GST-Cpa(247-370) polypeptide had been expressed in the following three different ways: expression in the vegetative cell, expression on the surface of the spore coat (fused to the CotB spore coat protein), and a combined approach of spore coat expression coupled with expression in the vegetative cell. Mice immunized orally or nasally with three doses of recombinant spores that carried GST-Cpa(247-370) on the spore surface showed the most striking responses. This included seroconversion with anti-Cpa(247-370)-specific immunoglobulin G (IgG) responses in their sera, a Th2 bias, and secretory IgA responses in saliva, feces, and lung samples. Neutralizing IgG antibodies to alpha toxin were detected using in vitro and in vivo assays, and a toxin challenge established protection. Mice immunized nasally or orally with recombinant spores were protected against a challenge with 12 median lethal doses of alpha toxin. Existing use of spores as competitive exclusion agents in animal feeds supports their use as a potentially economical and heat-stable vaccine for the poultry industry.
Hoang
Tran H
TH
School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey TW20 0EX, United Kingdom.
Hong
Huynh A
HA
Clark
Graeme C
GC
Titball
Richard W
RW
Cutting
Simon M
SM
D000284:Administration, Oral; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D000942:Antigens, Bacterial / Q000276:immunology; D001412:Bacillus subtilis / Q000235:genetics / Q000276:immunology*; D001427:Bacterial Toxins / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000276:immunology*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005738:Gas Gangrene / Q000517:prevention & control*; D051379:Mice; D008807:Mice, Inbred BALB C; D011201:Poultry Diseases / Q000517:prevention & control*; D010738:Type C Phospholipases / Q000235:genetics / Q000276:immunology*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D002135:Calcium-Binding Proteins; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens
Clin Microbiol Rev. 1996 Apr;9(2):216-34
8964036
J Immunol. 2004 Jan 15;172(2):1118-24
14707086
Infect Immun. 2006 Nov;74(11):6496-500
16923791
Microb Pathog. 2007 Oct;43(4):161-5
17604945
Vaccine. 2003 Oct 1;21(27-30):4215-24
14505901
FEMS Microbiol Lett. 1993 Jun 1;110(1):45-50
8319893
FEMS Microbiol Rev. 2005 Sep;29(4):813-35
16102604
Can J Comp Med. 1976 Jan;40(1):53-9
187291
Infect Immun. 2003 May;71(5):2810-8
12704155
Vaccine. 2004 Oct 22;22(31-32):4139-43
15474704
Vaccine. 1993 Sep;11(12):1253-8
8256506
Avian Dis. 2001 Jul-Sep;45(3):724-32
11569751
Vaccine. 2004 Mar 12;22(9-10):1177-87
15003646
Avian Pathol. 2004 Dec;33(6):537-49
15763720
Vaccine. 2007 Jan 4;25(2):356-65
16920233
Nat Biotechnol. 1997 Jul;15(7):653-7
9219268
Plasmid. 1984 Jul;12(1):1-9
6093169
Int J Antimicrob Agents. 2007 Aug;30(2):101-7
17467959
Clin Vaccine Immunol. 2007 Sep;14(9):1070-7
17634510
J Bacteriol. 2001 Nov;183(21):6294-301
11591673
J Immunol. 1987 May 15;138(10):3448-53
3106483
Vet Microbiol. 2003 Jul 17;94(3):245-56
12814892
Parasitol Res. 2008 Jan;102(2):293-7
17924143
Vaccine. 2007 Jan 4;25(2):346-55
17007969
Infect Immun. 1991 Dec;59(12):4338-42
1718874
Gene. 1996 Nov 21;180(1-2):57-61
8973347
Vet Microbiol. 2001 Mar 20;79(2):133-42
11230935
Vaccine. 2001 Jan 8;19(11-12):1460-6
11163669
J Infect Dis. 2004 Aug 15;190(4):767-73
15272405
Appl Environ Microbiol. 2005 Feb;71(2):968-78
15691955
Avian Dis. 1992 Jan-Mar;36(1):59-62
1567311
J Bacteriol. 2006 Apr;188(7):2692-700
16547057
J Immunol. 1996 Feb 15;156(4):1504-14
8568254
Vaccine. 2004 May 7;22(15-16):1873-85
15121298
Mol Microbiol. 1995 Jan;15(2):191-202
7746141
Vaccine. 2003 Jun 1;21 Suppl 2:S96-101
12763690
Clin Vaccine Immunol. 2008 May;15(5):805-16
18337376