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
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
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DOI：
10.1111/j.1348-0421.2007.tb03919.x
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发表时间：
2007-01-01
影响因子：
2.6
通讯作者：
Oguma, Keiji
中科院分区：
文献类型：
--
作者：
Lee, Jae-Chul;Hwang, Hyun-Jung;Oguma, Keiji
重组肉毒杆菌C型和D型神经毒素全重链（H, 100 kDa）及其n端（Hn, 50 kDa）和C端（Hc, 50 kDa）半片段在大肠杆菌中表达为谷胱甘肽s转移酶（GST）融合蛋白。消GST -将C型和D型的H （10 μ g）、Hn （5 μ g）、Hc （5 μ g）或Hn （5 μ g）和Hc （5 μ g）的混合物与等体积的佐剂混合，两次皮下注射。免疫后给小鼠注射10(6)个最小致死剂量(MLD)/0.5 ml的C或D毒素，其类型与免疫原相同。除Hn抗原外，其余抗原免疫小鼠均能抵抗10(5)~ 10(6)MLD/0.5 ml的毒素，而Hn抗原免疫小鼠则以100 MLD/0.5 ml的毒素死亡。C- hc和D- hc混合免疫小鼠，均为5 μ g，对C和D毒素均表现出高水平的抗性。与GST融合或消除GST制备免疫的抗体水平非常相似。这些结果表明，融合gst的重组Hc可以作为一种安全有效的动物C型和D型肉毒杆菌中毒疫苗。同样清楚的是，一次性接种大量的C-Hc或D-Hc， 100 μ g，对小鼠的疫苗试验是有用的。
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 mu g), Hn (5 mu g), Hc (5 mu g), or a mixture of Hn (5 mu g) and Hc (5 mu g) 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 mu g, 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 mu g, is useful for vaccine trials in mice.