MOLECULAR BASIS OF ANTIGENIC VARIATION ON MALARIA

疟疾抗原变异的分子基础

• 批准号: 8172319
• 负责人: MARY R GALINSKI
• 金额: $ 5.48万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172319
• 关键词: Agglutination; Antigenic Variation; Antigens; Attention; Biological Models; Blood; Cells; Chronic; Computer Retrieval of Information on Scientific Projects Database; Data; Down-Regulation; Funding; Gene Expression; Gene Expression Profile; Gene Proteins; Generations; Genetic; Genome; Grant; Human; Immune response; In Vitro; Infection; Institution; Macaca mulatta; Malaria; Manuscripts; Modeling; Molecular; Northern Blotting; Parasites; Phenotype; Plasmodium; Preparation; Proteins; Proteomics; Publishing; RNA; Regulation; Reporting; Research; Research Personnel; Resources; Reverse Transcriptase Polymerase Chain Reaction; Sampling; Source; Staging; Testing; Time; Transcript; United States National Institutes of Health; Variant; base; in vivo; insight; prototype; research study; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The objectives of this research are focused on the molecular mechanisms that govern variant antigen gene expression in Plasmodium. Antigenic variation is a fundamental adaptation to evade a host protective immune response and is one of the major factors contributing to the establishment of chronic blood infections. The classic P. knowlesi-rhesus monkey model is amenable to both in vitro and in vivo studies and unique stable clones of the P. knowlesi H strain expressing distinct SICA (Schizont Infected Cell Agglutination) variant antigen phenotypes after induced sequential switchings can be maintained after numerous in vivo passages (60 generations) in naive rhesus monkeys. These isogenic clonal lines provide a special tool for studies of the cellular and genetic mechanisms underlying clonal antigenic variation. We hypothesized that studies of the clonal phenotypes provide insights towards understanding the regulation of antigenic variation in vivo. This year, we published an important report on the redefined SICAvar prototype gene and protein, which emphasizes the need for further attention to the closure of the Pk genome data. We have also produced definitive data using proteomics (LC-MS/MS), real time RT-PCR and northern blot experiments defining the predominant transcripts and proteins produced by the classical P. knowlesi Pk1(A+)1+, Pk1(B+)1+ and Pk1(C+)1+ (i.e. SICA+) clones and show strong downregulation in the SICA- clones. Analyses are also underway with collaborators and a manuscript in preparation on a comprehensive P. knowlesi blood-stage transcriptome from RNA samples collected from an in vivo rhesus infection and in vitro adapted parasites. Proteomic data is also being generated, and compared with the transcriptome, with SICA expression and other hypotheses being tested. We also published a review emphasizing the importance of P. knowlesi not only as a model system, but also as a parasite shown recently to cause hundreds of cases of malaria in humans.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 本研究的目的是集中在疟原虫变异抗原基因表达的分子机制。抗原变异是逃避宿主保护性免疫应答的基本适应，并且是促成慢性血液感染建立的主要因素之一。经典的诺氏疟原虫-恒河猴模型适用于体外和体内研究，并且在诱导的顺序转换后表达不同SICA（裂殖体感染细胞凝集）变体抗原表型的诺氏疟原虫H株的独特稳定克隆可以在幼稚恒河猴中多次体内传代（60代）后保持。这些同基因克隆系为研究克隆抗原变异的细胞和遗传机制提供了一个特殊的工具。我们假设，克隆表型的研究提供了对了解体内抗原变异的调节的见解。 今年，我们发表了一份关于重新定义的SICAvar原型基因和蛋白质的重要报告，其中强调需要进一步关注Pk基因组数据的关闭。我们还使用蛋白质组学（LC-MS/MS）、真实的时间RT-PCR和北方印迹实验产生了明确的数据，这些实验定义了经典诺氏毕赤酵母Pk 1（A+）1+、Pk 1（B+）1+和Pk 1（C+）1+（即SICA+）克隆产生的主要转录物和蛋白质，并在SICA-克隆中显示出强烈的下调。 与合作者的分析也在进行中，并且正在准备一份关于从体内恒河猴感染和体外适应寄生虫收集的RNA样本中的全面P. knowlesi血液阶段转录组的手稿。蛋白质组数据也正在生成，并与转录组进行比较，SICA表达和其他假设正在测试中。 我们还发表了一篇综述，强调诺氏疟原虫不仅是一个模型系统，而且是一种寄生虫，最近在人类中引起了数百例疟疾病例。