Contemporary Circulating Enterovirus D68 Strains Infect and Undergo Retrograde Axonal Transport in Spinal Motor Neurons Independent of Sialic Acid

J Virol. 2019 Jul 30;93(16):e00578-19. doi: 10.1128/JVI.00578-19. Print 2019 Aug 15.


Enterovirus D68 (EV-D68) is an emerging virus that has been identified as a cause of recent outbreaks of acute flaccid myelitis (AFM), a poliomyelitis-like spinal cord syndrome that can result in permanent paralysis and disability. In experimental mouse models, EV-D68 spreads to, infects, and kills spinal motor neurons following infection by various routes of inoculation. The topography of virus-induced motor neuron loss correlates with the pattern of paralysis. The mechanism(s) by which EV-D68 spreads to target motor neurons remains unclear. We sought to determine the capacity of EV-D68 to spread by the neuronal route and to determine the role of known EV-D68 receptors, sialic acid and intracellular adhesion molecule 5 (ICAM-5), in neuronal infection. To do this, we utilized a microfluidic chamber culture system in which human induced pluripotent stem cell (iPSC) motor neuron cell bodies and axons can be compartmentalized for independent experimental manipulation. We found that EV-D68 can infect motor neurons via their distal axons and spread by retrograde axonal transport to the neuronal cell bodies. Virus was not released from the axons via anterograde axonal transport after infection of the cell bodies. Prototypic strains of EV-D68 depended on sialic acid for axonal infection and transport, while contemporary circulating strains isolated during the 2014 EV-D68 outbreak did not. The pattern of infection did not correspond with the ICAM-5 distribution and expression in either human tissue, the mouse model, or the iPSC motor neurons.IMPORTANCE Enterovirus D68 (EV-D68) infections are on the rise worldwide. Since 2014, the United States has experienced biennial spikes in EV-D68-associated acute flaccid myelitis (AFM) that have left hundreds of children paralyzed. Much remains to be learned about the pathogenesis of EV-D68 in the central nervous system (CNS). Herein we investigated the mechanisms of EV-D68 CNS invasion through neuronal pathways. A better understanding of EV-D68 infection in experimental models may allow for better prevention and treatment strategies of EV-D68 CNS disease.

Keywords: ICAM-5; enterovirus D68; motor neuron; retrograde transport; sialic acid.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Axonal Transport*
  • Cell Adhesion Molecules / metabolism
  • Central Nervous System Viral Diseases / metabolism
  • Central Nervous System Viral Diseases / virology
  • Disease Models, Animal
  • Enterovirus D, Human / physiology*
  • Enterovirus Infections / metabolism*
  • Enterovirus Infections / virology*
  • Host-Pathogen Interactions*
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Mice
  • Motor Neurons / cytology
  • Motor Neurons / metabolism*
  • Motor Neurons / virology*
  • Myelitis / metabolism
  • Myelitis / virology
  • N-Acetylneuraminic Acid / metabolism*
  • Nerve Tissue Proteins / metabolism
  • Neuromuscular Diseases / metabolism
  • Neuromuscular Diseases / virology
  • Paralysis / etiology


  • Cell Adhesion Molecules
  • ICAM5 protein, human
  • Nerve Tissue Proteins
  • N-Acetylneuraminic Acid

Supplementary concepts

  • acute flaccid myelitis