BIOLOGICAL EFFECTS OF PERTUSSIS TOXIN B-OLIGOMER

百日咳毒素 B-低聚物的生物学作用

• 批准号: 3146144
• 负责人: ERIK L HEWLETT
• 金额: $ 25.57万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3146144
• 关键词: ADP ribosylation; CD3 molecule; SDS polyacrylamide gel electrophoresis; T cell receptor; T lymphocyte; affinity labeling; calcium flux; cyclic AMP; cyclic GMP; cytotoxic T lymphocyte; glycolipids; glycoproteins; immune tolerance /unresponsiveness; immunoprecipitation; laboratory mouse; lymphocyte proliferation; monoclonal antibody; neoplastic cell culture for noncancer research; neurons; pertussis toxin; phosphatidylinositols; phospholipase A2; phospholipase C; platelet activation; prostaglandins; protein kinase C; protein structure function; protein tyrosine kinase; receptor binding; receptor expression; second messengers; site directed mutagenesis; thin layer chromatography; tissue /cell culture

Pertussis is a major cause of childhood morbidity in the developing world and as a result of concerns about pertussis vaccine reactogenicity, has increased in incidence in many areas of the developed world. Pertussis toxin (PT), a major virulence factor of Bordetella pertussis, elicits a variety of effects including lymphocytosis, sensitization to histamine, and enhancement of insulin secretion. Intriguingly, the B or binding component of PT (B-oligomer), which delivers its active portion to target cells, has biological effects of its own, namely calcium mobilization followed by lymphocyte proliferation and platelet aggregation. These actions, which are poorly understood, are independent of the ADP-ribosylation of G- proteins by which PT has the majority of its known effects. Acquisition of more information is important because many acellular pertussis vaccines contain PT in which holotoxin activities are eliminated, but B-oligomer effects are present or enhanced. In this proposal, different preparations of B-oligomer and holotoxin will be evaluated for potency in eliciting B- oligomer effects (lymphocyte proliferation and platelet activation). The second messenger systems involved will be explored and a screen of other cell types will be made for responses to B-oligomer. The receptor for B- oligomer action will be identified and the structural requirements for B- oligomer to elicit its effects will be studied with toxins containing mutations at specific sites. Finally, the relationship between B-oligomer mobilization of calcium and the subsequent intoxication of a target cell by ADP-ribosylation will be investigated. The resultant data will provide a perspective on using B-oligomer as a research tool and will guide its use as an antigen in acellular pertussis vaccine.

百日咳是发展中国家儿童发病的主要原因 由于对百日咳疫苗反应原性的担忧， 发达国家许多地区的发病率有所增加。 百日咳 毒素 (PT) 是百日咳博德特氏菌的主要毒力因子，可引起 多种效应，包括淋巴细胞增多、对组胺敏感等 增强胰岛素分泌。 有趣的是，B 或结合成分 PT（B-寡聚体）将其活性部分递送至靶细胞， 其自身的生物效应，即钙动员，然后 淋巴细胞增殖和血小板聚集。 这些行动， 知之甚少，独立于 G- 的 ADP-核糖基化 PT 发挥大部分已知作用的蛋白质。 收购 更多信息很重要，因为许多无细胞百日咳疫苗 含有消除全毒素活性的PT，但B-低聚物 效果存在或增强。 在这个提案中，不同的准备工作 将评估 B-寡聚物和全毒素在引发 B- 寡聚物效应（淋巴细胞增殖和血小板活化）。 这 将探索所涉及的第二信使系统，并筛选其他 细胞类型将针对 B 寡聚体做出反应。 B-受体 将确定低聚物的作用以及 B-的结构要求 将用含有的毒素来研究低聚物以引起其作用 特定位点的突变。 最后，B-低聚物之间的关系 钙的动员和随后的靶细胞中毒 ADP-核糖基化将被研究。 由此产生的数据将提供 使用 B-低聚物作为研究工具的观点并指导其使用 作为无细胞百日咳疫苗中的抗原。