Abstract
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 μg), Hn (5 μg), Hc (5 μg), or a mixture of Hn (5 μg) and Hc (5 μg) 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 106 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 105 to 106 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 μg, also showed a high level of resistance against both C and D toxins. Antibody levels immunized with GST fused- or GST eliminated-preparation 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 μg, is useful for vaccine trials in mice.
| Original language | English |
|---|---|
| Pages (from-to) | 445-455 |
| Number of pages | 11 |
| Journal | Microbiology and Immunology |
| Volume | 51 |
| Issue number | 4 |
| Publication status | Published - 2007 |
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Keywords
- Botulinum neurotoxin
- Botulism
- Clostridium botulinum
- Recombinant vaccine
ASJC Scopus subject areas
- Immunology and Microbiology(all)
- Microbiology (medical)
- Microbiology
Cite this
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. / Lee, Jae Chul; Hwang, Hyun Jung; Sakaguchi, Yoshihiko; Yamamoto, Yumiko; Arimitsu, Hideyuki; Tsuji, Takao; Watanabe, Toshihiro; Ohyama, Tohru; Tsuchiya, Tomofusa; Oguma, Keiji.
In: Microbiology and Immunology, Vol. 51, No. 4, 2007, p. 445-455.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - 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
AU - Lee, Jae Chul
AU - Hwang, Hyun Jung
AU - Sakaguchi, Yoshihiko
AU - Yamamoto, Yumiko
AU - Arimitsu, Hideyuki
AU - Tsuji, Takao
AU - Watanabe, Toshihiro
AU - Ohyama, Tohru
AU - Tsuchiya, Tomofusa
AU - Oguma, Keiji
PY - 2007
Y1 - 2007
N2 - 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 μg), Hn (5 μg), Hc (5 μg), or a mixture of Hn (5 μg) and Hc (5 μg) 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 106 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 105 to 106 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 μg, also showed a high level of resistance against both C and D toxins. Antibody levels immunized with GST fused- or GST eliminated-preparation 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 μg, is useful for vaccine trials in mice.
AB - 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 μg), Hn (5 μg), Hc (5 μg), or a mixture of Hn (5 μg) and Hc (5 μg) 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 106 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 105 to 106 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 μg, also showed a high level of resistance against both C and D toxins. Antibody levels immunized with GST fused- or GST eliminated-preparation 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 μg, is useful for vaccine trials in mice.
KW - Botulinum neurotoxin
KW - Botulism
KW - Clostridium botulinum
KW - Recombinant vaccine
UR - http://www.scopus.com/inward/record.url?scp=34247362076&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34247362076&partnerID=8YFLogxK
M3 - Article
C2 - 17446685
AN - SCOPUS:34247362076
VL - 51
SP - 445
EP - 455
JO - Microbiology and Immunology
JF - Microbiology and Immunology
SN - 0385-5600
IS - 4
ER -