Purification Toxoplasma gondii Tissue Cysts Using Percoll Gradients.
Purification Toxoplasma gondii Tissue Cysts Using Percoll Gradients.
复制标题
使用Percoll梯度纯化弓形虫组织囊肿。
DOI：
10.1002/cpmc.30
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发表时间：
2017-05-16
影响因子：
--
通讯作者：
Sinai AP
中科院分区：
文献类型：
--
作者：
Watts EA;Dhara A;Sinai AP
原生动物寄生虫弓形虫能够感染所有温血动物和人类。寄生虫的传染性、可传播形式包括最终猫科动物宿主内的性周期产生的卵囊和在所有慢性感染的温血宿主的无性周期期间在中枢神经系统和肌肉中形成弓形虫的组织包囊。这些组织囊肿中充满了生长缓慢的缓殖子，直到最近，这些缓殖子还被认为是在免疫充足的情况下处于休眠状态的实体。在免疫抑制期间重新激活活跃生长具有至关重要的临床意义。然而，我们对组织包囊或其所容纳的缓殖子知之甚少，因为组织包囊的多样性无法在细胞培养系统中复制。我们使用 Percoll 梯度对受感染小鼠大脑中的组织囊肿纯化进行优化，提供了一种有效的方法来回收体内来源的组织囊肿，可应用于成像、细胞生物学、生化、转录组和蛋白质组分析。
The protozoan parasite Toxoplasma gondii is capable of infecting all warm blooded animals and humans. Infectious, transmissible forms of the parasite include oocysts produced by the sexual cycle within the definitive feline host and tissue cysts that form Toxoplasma in the CNS and muscle during the asexual cycle within all chronically infected warm-blooded hosts. These tissue cysts are populated with slow growing bradyzoites which have been until recently thought to be dormant entities in the context of immune sufficiency. Reactivation to active growth during immune suppression is of critical clinical importance. Yet we know little about tissue cysts or the bradyzoites they house as the diversity of tissue cysts cannot be replicated in cell culture systems. Our optimization of tissue cyst purification from the brains of infected mice using Percoll gradients provides an efficient means to recover in vivo derived tissue cysts that can be applied to imaging, cell-biologic, biochemical, transcriptomic and proteomic analyses.