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: 3965842
• 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/3965842
• 关键词: Bordetella pertussis; aminoacid analyzer; antibacterial antibody; bacterial toxins; bactericidal immunity; bioassay; high performance liquid chromatography; immunological substance; monoclonal antibody; newborn animals; passive immunization; pertussis; thermostability; virulence

Bordetella pertussis produces several protein toxins including pertussis toxin (see project Z01 HD 1302) and dermonecrotic toxin or heat labile toxin (HLT). HLT, when injected subcutaneously into suckling mice, results in a local pronounced hemorrhagic lesion. The extent, localization, and nature of this toxin-induced injury suggests HLT contributes to the pathogenesis of B. pertussis. Conditions for assay of the toxin, using the suckling mouse model, have been established. Homogeneous preparations of HLT have been obtained and it has been established that the toxin is a single polypeptide chain with a molecular weight of about 150,000. The purified toxin has been used to produce antibodies. These antibodies have been used to document the purity of current toxin preparation. In addition, they neutralize toxic activity and high titer sera (1/20,000) have been obtained from immunized rabbits. Passive immunization of mice with HLT directed antibody protects mice against challenge with live B. pertussis suggesting that HLT may serve as a protective antigen. To investigate this possibility in greater detail, monoclonal antibodies to HLT are being produced. The toxicity of HLT is poorly understood and more detailed information establishing its role in pathogenesis is needed. Preliminary studies in which purified HLT was injected into mice indicate that the effects of the toxin are broad and histologic changes affect tissues such as spleen, liver, marrow, and thymus. Understanding of the pathologic changes in these tissues should contribute to elucidation of its mechanism of action and role in disease.

百日咳杆菌产生几种蛋白质毒素， 毒素（见项目Z 01 HD 1302）和皮肤坏死毒素或热不稳定毒素 毒素（HLT）。 当皮下注射给乳鼠时， 局部明显出血性损伤 范围、定位和 这种毒素诱导的损伤的性质表明HLT有助于 B的发病机制。百日咳。 毒素测定条件，使用 乳鼠模型。 均质制剂 已经获得了HLT，并且已经确定该毒素是一种 分子量约为150，000的单一多肽链。 纯化的毒素已被用于生产抗体。 这些抗体 已被用于记录当前毒素制剂的纯度。 在 此外，它们中和毒性活性和高滴度血清（1/20，000） 已从免疫的兔子获得。 小鼠被动免疫 HLT定向抗体保护小鼠免受活B的攻击。 提示HLT可作为保护性抗原。 到 为了更详细地研究这种可能性， HLT正在生产中。 对HLT的毒性了解甚少，需要提供更详细的信息 需要确定其在发病机制中的作用。 初步研究 将纯化的HLT注射到小鼠体内表明， 毒素是广泛的并且组织学变化影响组织如脾， 肝脏骨髓和胸腺 对乳腺癌病理变化的认识 这些组织有助于阐明其作用机制 在疾病中的作用。