Heparan sulfate in T.gondii attachment and invasion

描述(申请人提供)：弓形虫是一种广泛传播的 感染人类和动物的细胞内寄生虫。令人惊叹的能力 几乎感染每一种哺乳动物的弓形虫导致了一种假设，即它 识别硫酸乙酰肝素，一种广泛存在于多种 细胞。为了解决这一假设，我们提出了一系列研究，以解决 硫酸乙酰肝素在弓形虫与内皮细胞附着中的作用在目标1中，我们 建议检测弓形虫对突变和siRNA基因沉默的CHO的附着 在模拟血流的条件下HS生物合成中的细胞与 静态条件。此外，我们将利用肝素衍生物作为潜在的 阻断这些寄生虫附着的治疗剂。弓形虫可以引起 人类的疾病是通过跨越血脑屏障而发生的。因此，目标2将 研究HS在内皮细胞系和双层系统中的作用 血脑屏障。这一目标的目标是为房协在 弓形虫附着于内皮细胞组织。我们将使用CRE-loxP 重组系统在HS中创造内皮细胞基因敲除小鼠 生物合成，可以检测弓形虫的组织传播。此外， 这些组织可以被取出，并在体外检测弓形虫的附着。这个 这项研究的结果可能导致治疗NOT的治疗策略 只有弓形虫病，但有各种各样的寄生虫病和其他感染 利用硫酸乙酰肝素进入哺乳动物细胞。

DESCRIPTION (provided by applicant): Toxoplasma gondii is a wide-spread intracellular parasite infecting humans and animals. The amazing ability of Toxoplasma to infect almost every mammalian has led to the hypothesis that it recognizes heparan sulfate, a surface glyconjugate found on a wide variety of cells. To address this hypothesis, we propose a set of studies to addresses the role of heparan sulfate in Toxoplasma attachment to endothelium. In Aim 1, we propose to examine T gondii attachment to mutant and siRNA gene silenced CHO cells in HS biosynthesis under conditions that mimic blood flow as compared to static conditions. Further, we will utilize heparin derivatives as potential therapeutic agents to blockattachment of these parasites. Toxoplasma can cause disease in humans is by crossing the blood brain barrier. Thus, Aim 2 will examine the role of HS in endothelial cell lines and bi-layersystems that mimic the blood brain barrier. The goal of this aim is to establish a role for HS in Toxoplasma attachment to endothelial cell tissues. We will utilize Cre-loxP recombinant systems to create endothelial cell knockout mice in HS biosynthesis, which can be tested for Toxoplasma tissue dissemintion. Further, these tissues can be removed and assayed for T gondii attachment ex vivo. The results of this study could lead to therapeutic strategies for treating not only Toxoplasmosis, hut a wide variety of parasitemias and other infections that utilize heparan sulfate for entry into mammalian cells.