The JC Virus-Like Particle Overlay Assay

JC virus (JCV) belongs to the family of double-stranded DNA polyomaviruses and in humans causes a demyelinating disease of the central nervous system, progressive multifocal leukoencephalopathy (PML). It has been reported that sialic acids play a pivotal role in hemagglutination of red blood cells and entry into host cells of JCV and that JCV can enter a wide variety of cell types and localize to the nuclei. The outer shell of the JCV virion comprises the major capsid protein VP1, and a virus-like particle (VLP) consisting of recombinant VP1 made from Escherichia coli exhibit a virion-like structure and physiological functions (cellular attachment and intracytoplasmic trafficking) similar to those of JCV virions. To examine the mechanism of cell attachment of JCV, an overlay assay using a VLP has been developed, revealing that sialoglycoproteins, including α1 acid-glycoprotein, fetuin, and transferrin receptor bind with VLP. In addition, VLPs bind to glycolipids, such as lactosylceramide and gangliosides including GM3, GD2, GD3, GD1b, GT1b, and GQ1b, and VLP weakly bind to GD1a. In this section, detailed procedures for the synthesis of VLP from E. coli and VLP overlay assay are described.