BIOLOGY AND IMMUNOLOGY OF MALARIA PARASITE LIVER-STAGES

描述：我们对疟原虫肝期蛋白的了解主要是 一个黑盒子令人信服的证据表明，人类，非人类灵长类动物， 小鼠可以通过以下方式保护免受子孢子诱导的疟疾感染： 用辐射的子孢子免疫。肝脏阶段的研究很少 人类疟原虫的抗原已经进行，由于缺乏 合适的动物模型和这些抗原的有限供应。更好地 了解肝脏阶段疟原虫的生物学及其在 保护性免疫，我们建议分析寄生虫阶段特异性基因 在体外子孢子向裂殖子周期转变过程中的表达 和体内。为了便于EE阶段寄生虫的分析，我们将确定 约氏疟原虫基因在感染小鼠肝脏中的表达， 在感染后早期诱导，并在血液阶段周期期间关闭。 将利用差异显示分析来鉴定寄生虫基因， 在受照射和未受照射感染的肝细胞中表达 子孢子疟原虫肝脏阶段的寄生虫将通过激光捕获分离 显微切割及肝期cDNA文库的构建 构建和微阵列分析。此外，我们将识别疟原虫 在人中选择性表达的恶性疟原虫肝脏期特异性基因 体外和体内肝细胞。最初，感染的肝细胞获得 从移植了人肝细胞的hHGF/SCID和hHGF/RAG 1小鼠， 或肝细胞培养物，将用于产生恶性疟原虫EE阶段。 最后，我们将定义约氏疟原虫肝脏阶段基因产物的贡献 通过DNA免疫保护免疫。

DESCRIPTION: Our knowledge of Plasmodium liver stage proteins remains largely 'a black box'. Convincing evidence exists that humans, non-human primates, and mice can be protected against sporozoite-induced malaria infection by immunization with irradiated sporozoites. Few studies of the liver-stage antigens of human malaria parasites have been carried out due to the lack of appropriate animal models and the limited supply of these antigens. To better understand the biology of the liver-stage malaria parasite and its role in protective immunity, we propose to analyze the parasite stage-specific gene expression during the transition from sporozoite to merozoite cycle in vitro and in vivo. To facilitate the analysis of EE-stage parasites, we will identify Plasmodium yoelii gene in the infected mouse liver, the expression of which is induced early after infection, and shut off during the blood-stage cycle. Differential display analysis will be utilized to identify parasite genes that are expressed in hepatocytes infected with -irradiated and non-irradiated sporozoites. Plasmodium liver-stage schizonts will be isolated by laser-capture micro dissection and the generation of liver-stage cDNA for library construction and micro array analysis. In addition, we will identify Plasmodium falciparum liver-stage specific genes that are selectively expressed in human hepatocytes in vitro and in vivo. Initially, the infected hepatocytes obtained from either hHGF/SCID and hHGF/RAG1 mice transplanted with human hepatocytes, or hepatocyte cultures, will be used to generate P. falciparum EE-stages. Finally, we will define the contribution of P. yoelii liver-stage gene products in protective immunity via DNA immunizations.