Vaccination with recombinant whole heavy chain fragments of Clostridium botulinum Type C and D neurotoxins.
Mice and ducks were subcutaneously immunized with recombinant whole heavy (H) chains of Clostridium botulinum type C and D neurotoxins, which were expressed as glutathione S-transferase fusion proteins. In the case of mice, it was confirmed that two immunizations with type C- and D-H chains, 10 microg each time, significantly increased the specific antibodies against 100-kDa H chains of type C and D neurotoxins in an immunoblot analysis and an enzyme-linked immunosorbent assay, respectively. The mice immunized with type C- and D-H chains showed no symptoms of botulism when they were challenged with C- and D-16 S toxins at doses, given intraperitoneally, of up to 10(5) and 10(6) minmum lethal doses (MLD), respectively, per mouse. Ducks were immunized with a total of 100 microg of type C-H chain. The ducks also developed specific antibodies to the type C-H chain and showed significant protection against a challenge with 10(3) duck MLD of C-16 S toxin given intravenously. These results indicate that recombinant whole H chains can be used as an effective and safe vaccine for type C and D botulism in domestic animals.
Arimitsu
Hideyuki
H
Department of Bacteriology, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan. kuma@md.okayama-u.ac.jp
Lee
Jae-Chul
JC
Sakaguchi
Yoshihiko
Y
Hayakawa
Yuji
Y
Hayashi
Michiko
M
Nakaura
Miki
M
Takai
Hikaru
H
Lin
Song-Nan
SN
Mukamoto
Masafumi
M
Murphy
Tom
T
Oguma
Keiji
K
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000918:Antibody Specificity; D015153:Blotting, Western; D001905:Botulinum Toxins / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D001906:Botulism / Q000517:prevention & control; D003014:Clostridium botulinum / Q000737:chemistry / Q000235:genetics; D004372:Ducks; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D021122:Protein Subunits / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000276:immunology* / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D014611:Vaccination
D000907:Antibodies, Bacterial; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; C096325:botulinum toxin type D; D001905:Botulinum Toxins; C096324:botulinum toxin type C