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Analyses, using toxin fragment, of disturbances in autonomic nervous system in tetanus intoxication, a possible major cause of tetanus death

使用毒素片段分析破伤风中毒中自主神经系统的紊乱，这是破伤风死亡的可能主要原因

基本信息

批准号：
60480164
负责人：
MATSUDA Morihiro
金额：
$ 4.35万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

项目摘要

Fragments(A-B and C) of tetanus toxin were highly purified from mildly papain-digested toxin by HPLC-gel permeation and adsorption column chromatograpies. Fragment A-B preparation(200 <mu> g), further purified by affinity chromatography using anti-fragment C antibody as ligand, had no spastic toxicity which is typical for tetanus toxin, nor blocked neuromuscular transmission in rat, but showed definite, atypical toxicity("flaccid paralysis" or "weakness") when injected intramuscularly or intravenously into mice and killed them in 2-4 days. Unlike whole toxin, purified fragment A-B, blocked both inhibitory and excitatory synapses almost at the same time after injection into cat spinal cord. Toxin injected intramuscularly or intravenously into mice did not elicit hypertension but remarkable hypotension one to half day before death. Purified fragment A-B showed similar activity in mice. Toxin(0.64-2.6 ng) injected intravenously into rat, unlike toxin injected intramuscularly, elicited abnormal hypertension and subsequent death. Intracerebroventicular injection of toxin into rat gave more reproducible effects than intravenous injection. Immunocytochemical studies by indirect fluorescent technique showed localizatin of toxin in border between area postrema and nucleus solitarius in medulla oblongata. Further exact studies are being carried out using mouse monoclonal anti-bodies against toxin fragments. Toxin injected intravenously into rabbits (first 0.3 <mu> g and then 30 <mu> g 7 days later) showed remarkable fluctuations in blood pressure having repeated hypotensive phases resembling those in severe cases of tetanus patients. Control of blood pressure in these rabbits was attempted by intravenous administration of epinephine under total spinal anaesthesia. The method was successfully applied to treatment of tetanus patients.

用高效液相色谱-凝胶渗透法和吸附柱层析法从轻度木瓜酶消化的破伤风毒素中分离纯化出A-B和C片段。片段A-B制剂(200)经抗片段C抗体亲和层析纯化后，没有典型的破伤风毒素痉挛毒性，也没有阻断大鼠神经肌肉传递，但肌肉注射或静脉注射小鼠2-4天后表现出明确的非典型毒性(“松弛麻痹”或“虚弱”)。与完整毒素不同，纯化的片段A-B在注射到猫脊髓后几乎同时阻断抑制性和兴奋性突触。在小鼠死亡前1~半天，肌肉注射或静脉注射毒素不会引起高血压，但会引起明显的低血压。纯化的片段A-B在小鼠体内表现出类似的活性。与肌肉注射不同，静脉注射毒素(0.64-2.6 ng)可引起异常高血压和随后的死亡。大鼠脑室内注射毒素比静脉注射毒素具有更好的重复性。间接荧光免疫细胞化学研究表明，毒素定位于延髓最后区与孤束核交界处。进一步的精确研究正在使用针对毒素片段的鼠单抗进行。给兔静脉注射毒素(先0.3mt；gt；g，然后7d后30lt；mu&gt；g)可见血压显著波动，反复出现类似破伤风重症患者的低血压时相。在全脊髓麻醉下，通过静脉注射肾上腺素来控制这些兔的血压。该方法已成功应用于破伤风患者的治疗。