Production and evaluation of a recombinant chimeric vaccine against clostridium botulinum neurotoxin types C and D.
Bovine botulism is a fatal disease that is caused by botulinum neurotoxins (BoNTs) produced by Clostridium botulinum serotypes C and D and that causes great economic losses, with nearly 100% lethality during outbreaks. It has also been considered a potential source of human food-borne illness in many countries. Vaccination has been reported to be the most effective way to control bovine botulism. However, the commercially available toxoid-based vaccines are difficult and hazardous to produce. Neutralizing antibodies targeted against the C-terminal fragment of the BoNT heavy chain (HC) are known to confer efficient protection against lethal doses of BoNTs. In this study, a novel recombinant chimera, consisting of Escherichia coli heat-labile enterotoxin B subunit (LTB), a strong adjuvant of the humoral immune response, fused to the HC of BoNT serotypes C and D, was produced in E. coli. Mice vaccinated with the chimera containing LTB and an equivalent molar ratio of the chimera without LTB plus aluminum hydroxide (Al(OH)3) developed 2 IU/mL of antitoxins for both serotypes. Guinea pigs immunized with the recombinant chimera with LTB plus Al(OH)3 developed a protective immune response against both BoNT/C (5 IU/mL) and BoNT/D (10 IU/mL), as determined by a mouse neutralization bioassay with pooled sera. The results achieved with guinea pig sera fulfilled the requirements of commercial vaccines for prevention of botulism, as determined by the Brazilian Ministry of Agriculture, Livestock and Food, Supply. The presence of LTB was essential for the development of a strong humoral immune response, as it acted in synergism with Al(OH)3. Thus, the vaccine described in this study is a strong candidate for the control of botulism in cattle.
Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Luciana A F
LA
Gil
Carlos Eduardo P
CE
da Cunha
Gustavo M S G
GM
Moreira
Felipe M
FM
Salvarani
Ronnie A
RA
Assis
Francisco Carlos F
FC
Lobato
Marcelo
M
Mendonça
Odir A
OA
Dellagostin
Fabricio R
FR
Conceição
D000536:Aluminum Hydroxide / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D000992:Antitoxins / Q000276:immunology; D001427:Bacterial Toxins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D001905:Botulinum Toxins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D001906:Botulism / Q000276:immunology* / Q000517:prevention & control / Q000662:veterinary; D002417:Cattle; D047968:Clostridium botulinum type C / Q000276:immunology*; D047969:Clostridium botulinum type D / Q000276:immunology*; D004341:Drug Evaluation; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D029968:Escherichia coli Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005260:Female; D006168:Guinea Pigs; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014611:Vaccination / Q000379:methods; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology* / Q000592:standards
D000907:Antibodies, Bacterial; D000992:Antitoxins; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D000536:Aluminum Hydroxide; C027538:heat-labile enterotoxin, E coli; D001905:Botulinum Toxins