BIOLOGY AND IMMUNOLOGY OF MALARIA PARASITE LIVER-STAGES

描述：我们对疟原虫肝脏阶段蛋白的了解仍然很多。 “一个黑匣子”。有令人信服的证据表明，人类、非人类灵长类和 可以通过以下方法保护小鼠不受子孢子引起的疟疾感染 用辐照的子孢子进行免疫。关于肝期的研究很少 人类疟疾寄生虫的抗原已经进行了，因为缺乏 合适的动物模型和这些抗原的有限供应。为了更好地 了解肝期疟疾寄生虫的生物学及其在 保护性免疫，我们建议分析寄生虫阶段特异性基因 体外子孢子向裂殖子周期转换过程中的表达 在活体内。为了方便分析EE阶段的寄生虫，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，在血液期周期关闭。 差异显示分析将被用来鉴定寄生虫基因 在辐照和未辐照感染的肝细胞中表达 子孢子。用激光捕获法分离肝期疟原虫裂殖体 显微切割与肝期文库的构建 构建和微阵列分析。此外，我们还将鉴定疟原虫 恶性疟原虫肝期特异基因在人中的选择性表达 体外和体内肝细胞。最初，受感染的肝细胞获得 从hHGF/SCID和hHGF/RAG1小鼠移植人肝细胞， 或肝细胞培养，将用于产生恶性疟原虫EE期。 最后，我们将确定约氏疟原虫肝期基因产物的贡献。 通过DNA免疫进行保护性免疫。

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.