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

• Published in: Microbiology and immunology 2007-01, Vol.51 (4), p.445-455
• Main Authors: Lee, Jae-Chul, Hwang, Hyun-Jung, Sakaguchi, Yoshihiko, Yamamoto, Yumiko, Arimitsu, Hideyuki, Tsuji, Takao, Watanabe, Toshihiro, Ohyama, Tohru, Tsuchiya, Tomofusa, Oguma, Keiji
• Format: Article
• Language: English
• Subjects: Animals; Bacterial Vaccines - immunology; botulinum neurotoxin; Botulinum Toxins - genetics; Botulinum Toxins - immunology; Botulinum Toxins - pharmacology; botulism; Botulism - prevention & control; Clostridium botulinum; Clostridium botulinum type C - chemistry; Clostridium botulinum type D - chemistry; Escherichia coli; Escherichia coli - genetics; Glutathione Transferase - metabolism; Mice; Molecular Weight; Neurotoxins; Protein Subunits - administration & dosage; Protein Subunits - genetics; Protein Subunits - immunology; Recombinant Fusion Proteins - administration & dosage; Recombinant Fusion Proteins - immunology; Recombinant Fusion Proteins - isolation & purification; recombinant vaccine; Toxicity Tests; Vaccination; Vaccines, Synthetic - immunology
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.2007.tb03919.x

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.