Utilization of sialic acid as a coreceptor is required for reovirus-induced biliary disease
Erik S. Barton, … , J. Denise Wetzel, Terence S. Dermody
Erik S. Barton, … , J. Denise Wetzel, Terence S. Dermody
Published June 15, 2003
Citation Information: J Clin Invest. 2003;111(12):1823-1833. https://doi.org/10.1172/JCI16303.
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Utilization of sialic acid as a coreceptor is required for reovirus-induced biliary disease

Abstract

Infection of neonatal mice with some reovirus strains produces a disease similar to infantile biliary atresia, but previous attempts to correlate reovirus infection with this disease have yielded conflicting results. We used isogenic reovirus strains T3SA– and T3SA+, which differ solely in the capacity to bind sialic acid as a coreceptor, to define the role of sialic acid in reovirus encephalitis and biliary tract infection in mice. Growth in the intestine was equivalent for both strains following peroral inoculation. However, T3SA+ spread more rapidly from the intestine to distant sites and replicated to higher titers in spleen, liver, and brain. Strikingly, mice infected with T3SA+ but not T3SA– developed steatorrhea and bilirubinemia. Liver tissue from mice infected with T3SA+ demonstrated intense inflammation focused at intrahepatic bile ducts, pathology analogous to that found in biliary atresia in humans, and high levels of T3SA+ antigen in bile duct epithelial cells. T3SA+ bound 100-fold more efficiently than T3SA– to human cholangiocarcinoma cells. These observations suggest that the carbohydrate-binding specificity of a virus can dramatically alter disease in the host and highlight the need for epidemiologic studies focusing on infection by sialic acid–binding reovirus strains as a possible contributor to the pathogenesis of neonatal biliary atresia.

Authors

Erik S. Barton, Bryan E. Youree, Daniel H. Ebert, J. Craig Forrest, Jodi L. Connolly, Tibor Valyi-Nagy, Kay Washington, J. Denise Wetzel, Terence S. Dermody

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Figure 3

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Genotypic and phenotypic characterization of virus isolated from mice fo...
Genotypic and phenotypic characterization of virus isolated from mice following infection with T3SA– and T3SA+. (a) Viral genomic RNA was extracted from either homogenized livers of infected mice or second-passage lysate stocks. S1 gene segment cDNA’s were amplified using RT-PCR (labeled at right as Primary and Secondary), and restriction enzyme digestion was performed using BstNI. Digested secondary PCR products (labeled at right as 355 bp and 190 bp) were visualized by agarose gel electrophoresis and ethidium bromide staining. Undigested (U) and digested (D) stock virions are shown along with infected livers from days 4, 8, and 12 after inoculation. The lanes labeled M were loaded with 100-bp markers. (b) Homogenized livers obtained 4, 8, and 12 days after inoculation were passaged in L cells for 72 hours. L cell lysate (150 μl) was adsorbed to either L cells or MEL cells (2 × 105) and incubated at 37°C for 24 hours. Viral titers at 0 hours and 24 hours were determined from liver samples obtained from three to four mice by plaque assay using L cells. Viral yields were calculated by dividing titer at 24 hours by titer at 0 hours, and average viral yields for all at 3 days after inoculation are shown. Error bars indicate SEM.