Autophagy and ER stress during varicella infection

自噬最近在水痘感染的培养细胞中得到证实， 带状疱疹病毒（VZV）以及人带状疱疹囊泡中。点状自噬体的存在 在带状疱疹皮肤囊泡中，自噬（大自噬）是一种相关的生物学机制， 在由这种疱疹病毒引起的疾病的自然史过程中。重要的是， 进行了充分的初步研究，以证明VZV感染诱导的自噬 不像HSV诱导的自噬。首先，VZV基因组缺乏与VZV基因组的同源物。 HSV ICP 34.5基因。其次，我们现在已经通过两种不同的方法表明，VZV感染 快速诱导显著的ER应激，这是HSV系统中不知道发生的事件。因此 重申该建议的中心假设是ER应激是VZV的关键组成部分 感染时，ER应激与ER中过量错误折叠VZV糖蛋白有关， ER相关的降解和自噬是缓解ER应激的结果。这些 这些事件允许受感染的细胞存活更长时间，以避免凋亡，并且在这个意义上可以是 被认为是前病毒。检验这一假设的研究计划包括三个具体目标。目的 1被称为自噬的诱导作为VZV诱导的ER应激的结果。目标2被称为 VZV诱导的内质网应激激活未折叠蛋白反应（UPR）。目标3： 涉及A.阿尔文实验室，包括研究自噬和ER应激在SCID-hu 小鼠模型，感染各种重组突变VZV基因组后。在我们 研究计划，我们将继续扩大实验方法之前的调查 针对从ER应激到UPR的三个传感器通路（IRE 1、ATF 6和 PERK）。我们已经在VZV感染的细胞中检测到四种与UPR相关的蛋白， 包括BiP（HSPA 5）、HSPA 8、HSPD 1和PPIA：肽基丙基顺反异构酶）。更 最近我们已经在VZV感染的细胞中检测到剪接形式的XBP 1（X-Box结合蛋白）， 普遍定期审议的另一个标志。最终的方法将包括动物实验， 制备含有大量gE突变的重组病毒。这些突变体的表型 已经在细胞培养物和SCID小鼠模型中确定了病毒。因此 上述实验计划将确定ER应激和自噬在 VZV感染的细胞反应。这些结果不仅与我们对 水痘疫苗接种，而且还削弱带状疱疹相关的疾病称为带状疱疹后神经痛。

Autophagy has recently been demonstrated in both cultured cells infected with varicella- zoster virus (VZV) as well as in a human zoster vesicle. The presence of punctate autophagosomes in a zoster skin vesicle firmly established that autophagy (macroautophagy) is a relevant biological process during the natural history of disease caused by this herpesvirus. Of importance, we have carried out sufficient preliminary studies to document that autophagy induced by VZV infection does not resemble HSV-induced autophagy. First of all, the VZV genome lacks a homolog of the HSV ICP 34.5 gene. Secondly, we have now shown by two different methods that VZV infection quickly induces marked ER stress, an event not known to occur in the HSV system. Therefore, the re-stated central hypothesis for this proposal is that ER stress is a critical component of VZV infection, that ER stress is related to an over abundance of misfolded VZV glycoproteins in the ER, and that ER-associated degradation and autophagy are consequences that relieve ER stress. These events allow the infected cell to survive longer, to avoid apoptosis, and in that sense can be considered pro-viral. The Research Plan to test this hypothesis involves three Specific Aims. Aim 1 is called Induction of autophagy as a consequence of VZV induced ER stress. Aim 2 is called Activation of the unfolded protein response (UPR) by VZV induced ER stress. Aim 3, which involves the A. Arvin laboratory, includes Studies of autophagy and ER stress in the SCID-hu mouse model, following infection with various recombinant mutated VZV genomes. In our Research Plan, we will continue prior investigations with an expanded experimental approach directed toward the three sensor pathways that lead from ER stress to the UPR (IRE1, ATF6 and PERK). We have already detected four proteins associated with the UPR in VZV infected cells, including BiP (HSPA5), HSPA8, HSPD1 and PPIA: peptidyl-propyl-cis-trans-isomerase). More recently we have detected the spliced form of XBP1 (X-Box binding protein) in VZV infected cells, another marker of the UPR. The final approach will include animal experiments with already prepared recombinant viruses containing numerous gE mutations. The phenotypes of these mutant viruses have already been determined in both cell culture and the SCID mouse model. Thus the above experimental plan will determine the relative importance of ER stress and autophagy in the cellular response to VZV infection. The results are relevant not only to our understanding of varicella vaccination but also the debilitating zoster-related illness called post-herpetic neuralgia.