PERTUSSIS HEAT LABILE TOXIN (HLT)--ISOLATION AND CHARACTERIZATION

百日咳热不稳定毒素 (HLT)——分离和表征

• 批准号: 4693834
• 负责人: R D SEKURA
• 金额: --
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/4693834
• 关键词: Bordetella pertussis; adenylate cyclase; aminoacid analyzer; antibacterial antibody; bacterial toxins; bactericidal immunity; bioassay; high performance liquid chromatography; immunological substance; newborn animals; pertussis; proteolysis; thermostability; virulence; zymogens

Bordetella pertussis produces several protein toxins including pertussis toxin and dermonecrotic toxin (heat labile toxin or HLT). HLT, injected subcutaneously into suckling mice, induces a lcoal pronounced hemorrhagic lesion. The extent, localization, and nature of this toxin-induced injury suggests HLT contributes to te pathogenesis of B. pertussis. Conditions for assay of the toxin using the suckling mouse model have been established. By use of conventional protein purification techniques and high pressure liquid chromatography, homogeneous preparations of HLT have been obtained. These studies show that the toxin is a single polypeptide chain with a molecular weight of about 150,000. The purified toxin has been subjected to amino acid analysis and has been used to produce antibodies. These antibodies have been used to document the purity of current toxin preparation. In addition, HLT antibodies neutralize toxic activity and high titer sera (1/20,000) have been obtained from immunized rabbits. HLT, in addition to mediating a dermonecrotic lesion, was shown to be lethal, induce a short term elevation (3 days) of white blood cells, and elicit and marked atrophy and "bleaching" of the spleen. The significance of these changes to possible biochemical mechanisms of action and their relation to the pathogenesis of pertussis are being investigated.

百日咳博德特氏菌产生多种蛋白质毒素，包括百日咳 毒素和皮肤坏死毒素（不耐热毒素或 HLT）。 HLT，注入 皮下注射到乳鼠体内，引起明显的出血 病变。 这种毒素引起的损伤的程度、定位和性质 表明 HLT 有助于百日咳博德特氏菌的发病机制。 状况 使用乳鼠模型进行毒素测定 已确立的。 通过使用常规的蛋白质纯化技术和 高压液相色谱法，HLT均质制剂具有 已获得。 这些研究表明该毒素是单一多肽 链，分子量约为150,000。 纯化后的毒素已进行氨基酸分析并已 用于产生抗体。 这些抗体已被用来记录 当前毒素制剂的纯度。 此外，HLT 抗体 中和毒性活性并获得高滴度血清（1/20,000） 来自免疫兔子。 HLT，除了介导皮肤坏死之外 病变，被证明是致命的，引起短期升高（3天） 白细胞，并引起明显的萎缩和“漂白” 脾。 这些变化对可能的生化的意义 作用机制及其与百日咳发病机制的关系 正在接受调查。