EV-D68是一种不是脊髓灰质炎病毒的肠道病毒，与严重的呼吸道疾病和急性松弛性脊髓炎(AFMD)有关，AFMD是一种类似脊髓灰质炎的疾病，可导致儿童瘫痪。自2014年以来，与EV-D68的流行和基因多样性增加有关的AFMD爆发情况不断增加，尽管该病毒在20世纪60年代被发现。EV-D68如何靶向中枢神经系统的机制尚不清楚。由于肠道病毒是不会常规感染其他动物物种的人类病原体，建立一个人类中枢神经系统模型对于了解发病机理非常重要。在这里，我们描述了两种基于人类脊髓器官样体(hSCO)的EV-D68感染模型，该器官样体来源于诱导多能干细胞(iPSC)系列。一个hSCO模型主要由脊髓运动神经元组成，而另一个模型包括多种神经细胞系谱，包括运动神经元、中间神经元和胶质细胞。这些hSCO可被现代株的EV-D68有效感染，而历史型株则不行，并且能够在至少2周的时间内产生细胞外病毒而没有明显的细胞病变影响。相比之下，与hSCO感染另一种肠道病毒Echo11的情况相比，会导致明显的结构破坏和细胞凋亡。综上所述，这些发现表明在具有AFMD的脊髓中，EV-D68感染不是神经细胞死亡的唯一介导因素，免疫反应引起的继发损伤可能是发病机理的贡献因素。重要性AFMD是一种罕见疾病，会导致患儿长期后遗症。目前尚不清楚EV-D68是如何导致儿童瘫痪的，并且没有有效的治疗和预防策略。小鼠不是EV-D68的天然宿主，因此，现有的小鼠模型使用免疫抑制或新生小鼠、小鼠适应病毒或颅内接种。为了补充现有模型，我们报道了两个EV-D68感染的hSCO模型。这些三维的多细胞模型由人类细胞组成，并包括多种神经系谱，包括运动神经元，中间神经元和胶质细胞。这些新的EV-D68感染的hSCO模型将有助于理解EV-D68如何损伤人类脊髓，从而可能导致对该病毒的新治疗和预防策略。

Enterovirus D68 (EV-D68) is a nonpolio enterovirus associated with severe respiratory illness and acute flaccid myelitis (AFM), a polio-like illness causing paralysis in children. AFM outbreaks have been associated with increased circulation and genetic diversity of EV-D68 since 2014, although the virus was discovered in the 1960s. The mechanisms by which EV-D68 targets the central nervous system are unknown. Since enteroviruses are human pathogens that do not routinely infect other animal species, establishment of a human model of the central nervous system is essential for understanding pathogenesis. Here, we describe two human spinal cord organoid (hSCO)-based models for EV-D68 infection derived from induced, pluripotent stem cell (iPSC) lines. One hSCO model consists primarily of spinal motor neurons, while the another model comprises multiple neuronal cell lineages, including motor neurons, interneurons, and glial cells. These hSCOs can be productively infected with contemporary strains, but not a historic strain, of EV-D68 and produce extracellular virus for at least 2 weeks without appreciable cytopathic effect. By comparison, infection with hSCO with another enterovirus, echovirus 11, causes significant structural destruction and apoptosis. Together, these findings suggest that EV-D68 infection is not the sole mediator of neuronal cell death in the spinal cord in those with AFM and that secondary injury from the immune response likely contributes to pathogenesis. IMPORTANCE AFM is a rare condition that causes significant morbidity in affected children, often contributing to life-long sequelae. It is unknown how EV-D68 causes paralysis in children, and effective therapeutic and preventative strategies are not available. Mice are not native hosts for EV-D68, and thus, existing mouse models use immunosuppressed or neonatal mice, mouse-adapted viruses, or intracranial inoculations. To complement existing models, we report two hSCO models for EV-D68 infection. These three-dimensional, multicellular models comprised human cells and include multiple neural lineages, including motor neurons, interneurons, and glial cells. These new hSCO models for EV-D68 infection will contribute to understanding how EV-D68 damages the human spinal cord, which could lead to new therapeutic and prophylactic strategies for this virus.