Nonhuman Primate Model to Assess Fetal Zika Virus Infection Complications

摘要 感染寨卡病毒(ZIKV)最近被认为与以下疾病的发病率增加有关 巴西的小头畸形症，然而，ZIKV在胎儿中存在对神经发育的实际影响， 包括小头畸形症，人们对此知之甚少。虽然正在进行的流行病学很可能会加强怀孕 结果与寨卡病毒相关，并可能揭示影响母体感染的动态 在人类身上，直接询问影响胎儿结局的因素是不可能的 临床环境。非人灵长类动物提供了一个极好的机会来洞察这些 ZIKV感染胎儿的病理生理过程我们的总体目标是建立胎儿寨卡病毒感染 以羊水分娩为模型，研究其对胎儿生长发育和中枢神经系统发育的影响。通过直接感染 携带病毒的胎儿，我们将绕过经胎盘传播的潜在瓶颈，这可能是一种 研究胎儿结局的限速步骤，因为在大多数情况下，胎盘可以预防，而不是 促进病原体向胎儿的传播。因此，将病毒直接引入羊水中将 是一个更有效的方法来定义病理事件直接在胎儿间隔室。我们建议两个 具体目标是解决我们的目标。 具体目的1.为了确定猕猴胚胎对ZIKV感染是否敏感，我们将绕过 潜在的胎盘屏障并直接接种羊水，监测胎儿的健康和发育 病毒学、免疫学、超声波和核磁共振分析。 具体目的2.为了明确胎儿感染的个体发育和组织趋向性，我们将顺序收集胎儿 宫内感染后的组织图谱病毒嗜性和胎儿发病机制，重点全面 胎儿组织病理学。 该模型的开发和验证将建立一种快速筛查胎儿感染对 中枢神经系统以及整个胎儿发育。非人灵长类的直接验证 研究胎儿神经病理、生长受限和可能的胎儿死亡的模型将是非常有价值的 在建立非人灵长类平台方面向前迈进一步，以获得对严重 巴西疫情中出现的神经病理学。可预测的胎儿结局将允许选择性滴定 通过组织病理学判断与剂量相关的神经影响，使模型也会揭示出更细微的 神经行为的侮辱比小头畸形，很可能是一个极端的结果。

ABSTRACT Infection with Zika virus (ZIKV) has been very recently implicated in an increase in the incidence of microcephaly in Brazil, however the actual impact of the presence of ZIKV in the fetus on neural development, including microcephaly, is poorly understood. While ongoing epidemiology will likely strengthen pregnancy outcome associations with ZIKV and possibly reveal the dynamics of maternal infection that influence pregnancy outcome, direct interrogation of factors that drive fetal outcomes will not be possible in human clinical settings. The nonhuman primate offers an outstanding opportunity to gain insight into these pathophysiological processes in fetal infection with ZIKV. Our overall goal is to establish a fetal ZIKV infection model by amniotic fluid delivery to study its effects on fetal growth and CNS development. By directly infecting the fetus with the virus, we will bypass the potential bottleneck of transplacental transmission, which may be a rate-limiting step in studying fetal outcomes, since in most circumstances the placenta prevents, and does not facilitate, transmission of pathogens to the fetus. Thus, direct introduction of the virus into the amniotic fluid will be a more efficient approach for defining pathological events directly in the fetal compartment. We propose two Specific Aims to address our goal. Specific Aim 1. To determine whether the macaque fetus is sensitive to ZIKV infection, we will bypass the potential placental barrier and directly inoculate the amniotic fluid, monitoring fetal well-being and development with virological, immunological, ultrasound and MRI analyses. Specific Aim 2. To define the ontogeny and tissue tropism of fetal infection, we will sequentially collect fetal tissues following intrauterine infection to map viral tropism and fetal pathogenesis, focusing on comprehensive fetal histopathology. The development and validation of this model will establish a rapid screen for the impact of fetal infection on the central nervous system as well as overall fetal development. Direct validation of the nonhuman primate model to study fetal neuropathology, growth restriction, and possibly fetal demise will be an incredibly valuable step forward in building the nonhuman primate platform to obtain a mechanistic understanding of the severe neuropathology seen in the outbreak in Brazil. A predictable fetal outcome will allow selective titration of the dose-related neurological impact as judged by histopathology, so that the model will also reveal more subtle neurobehavioral insults than microcephaly, likely to be an extreme outcome.