Creation of a tissue culture model for Toxoplasma gondii sexual development

描述（由申请人提供）：刚地弓形虫的性循环仅限于猫的肠道，在那里形成数百万个卵囊并随后随粪便排出。这个建议的目的是概括弓形虫的性周期在组织培养。体外性发育条件的创造将允许对特定基因对生物过程和冗余途径的贡献进行快速分析。由于弓形虫的无性发育已经可以在组织培养中进行，因此它也将允许对原体寄生虫的完整生命周期进行分子分析。弓形虫的组织培养性发育的另一个好处是稳定地生产弓形虫疫苗载体，以对抗自身和其他病原体，如疟原虫。表达约氏疟原虫或诺氏疟原虫环孢子子蛋白的重组弓形虫分别在小鼠和恒河猴中提供抗疟疾感染的保护性免疫。作为卵囊生产的弓形虫疫苗载体将是理想的，因为它在几乎任何环境条件下都是稳定的，而且它将是口服接种。我的实验室也在鉴定对弓形虫形成慢性感染至关重要的毒力基因。这些建议的协同工作将使我们能够创造一种减毒疫苗株，可以从细胞培养中产生稳定的卵囊。公共卫生相关性：刚地弓形虫的性循环仅限于猫的肠道，在那里形成数百万个卵囊，随后随粪便排出。这个建议的目的是概括弓形虫的性周期在组织培养。弓形虫的组织培养性发育的另一个好处是稳定地生产弓形虫疫苗载体，以对抗自身和其他病原体，如疟原虫。

DESCRIPTION (provided by applicant): The sexual cycle of Toxoplasma gondii is limited to the feline intestine where millions of oocysts are formed and subsequently excreted in their feces. The goal of this proposal is to recapitulate the T. gondii sexual cycle in tissue culture. Creation of ex vivo sexual development conditions will allow the contribution of specific genes to biological processes and redundant pathways to be rapidly analyzed. It will also allow for a molecular analysis of the complete lifecycle of a protistan parasite, as T. gondii asexual development can already be performed in tissue culture. Another benefit of tissue culture sexual development of T. gondii will be the stable production of a T. gondii vaccine vector against itself and other pathogens, such as Plasmodium. Recombinant T. gondii expressing circumsporozoite protein from Plasmodium yoelii or P. knowlesi provides protective immunity against malaria infection in mice and rhesus monkeys, respectively. A T. gondii vaccine vector produced as an oocyst will be ideal because it will be stable in virtually any environmental condition and it will be an oral inoculation. My laboratory is also identifying the virulence genes critical for T. gondii to establish a chronic infection. Synergizing the work from these proposals will allow us to create an attenuated vaccine strain that can be produced as a stable oocyst from cell culture. PUBLIC HEALTH RELEVANCE: The sexual cycle of Toxoplasma gondii is limited to the feline intestine where millions of oocysts are formed and subsequently excreted in their feces. The goal of this proposal is to recapitulate the T. gondii sexual cycle in tissue culture. Another benefit of tissue culture sexual development of T. gondii will be the stable production of a T. gondii vaccine vector against itself and other pathogens, such as Plasmodium.