C terminal half fragment (50 kDa) of heavy chain components of Clostridium botulinum type C and D neurotoxins can be used as an effective vaccine.
Recombinant whole heavy chains (H, 100 kDa) and their N-terminal (Hn, 50 kDa) and C-terminal (Hc, 50 kDa) half fragments of Clostridium botulinum type C and D neurotoxins were expressed as glutathione S-transferase (GST) fusion proteins in Escherichia coli. GST eliminated-preparations of H (10 microg), Hn (5 microg), Hc (5 microg), or a mixture of Hn (5 microg) and Hc (5 microg) of types C and D were mixed with an equal volume of adjuvant, and then were twice injected into mice subcutaneously. After immunization, the mice were challenged with up to 10(6) the minimum lethal doses (MLD)/0.5 ml of C or D toxin, the type of which was same as that of the immunogens. All of the mice immunized with antigens except for Hn survived against 10(5) to 10(6) MLD/0.5 ml of the toxins, but the mice immunized with Hn were killed by 100 MLD/0.5 ml. The mice immunized with a mixture of C-Hc and D-Hc, each 5 microg, also showed a high level of resistance against both C and D toxins. Antibody levels immunized with GST fused-or GST eliminatedpreparation were quite similar. These results indicate that recombinant GST-fused Hc can be used as a safe and effective vaccine for type C and D botulism in animals. It also became clear that one time inoculation with a large amount of C-Hc or D-Hc, 100 microg, is useful for vaccine trials in mice.
Lee
Jae-Chul
JC
Department of Bacteriology, Okayama University Medical School, Okayama, Japan.
Hwang
Hyun-Jung
HJ
Sakaguchi
Yoshihiko
Y
Yamamoto
Yumiko
Y
Arimitsu
Hideyuki
H
Tsuji
Takao
T
Watanabe
Toshihiro
T
Ohyama
Tohru
T
Tsuchiya
Tomofusa
T
Oguma
Keiji
K
D000818:Animals; D001428:Bacterial Vaccines / Q000276:immunology*; D001905:Botulinum Toxins / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D001906:Botulism / Q000517:prevention & control; D047968:Clostridium botulinum type C / Q000737:chemistry*; D047969:Clostridium botulinum type D / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D009498:Neurotoxins; D021122:Protein Subunits / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000276:immunology* / Q000302:isolation & purification; D018675:Toxicity Tests; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000276:immunology
D001428:Bacterial Vaccines; D009498:Neurotoxins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D005982:Glutathione Transferase; D001905:Botulinum Toxins