High-throughput automated image analysis of neuroinflammation and neurodegeneration enables quantitative assessment of virus neurovirulence
Section snippets
Viruses
LGTV wild-type strain TP21 was received from the Rockefeller Foundation Collection and amplified in Vero cells [31]. The chimeric recombinant TBEV/DEN4Δ30 virus, containing the prM and E genes of the TBEV strain Sofjin and a 30-nucleotide deletion in the 3′ non-coding region of the genome, was originally recovered after transfection of Vero cells with RNA transcripts of its full-length chimeric cDNA genome [32]. The YF 17D vaccine virus was received from Sanofi Pasteur, Inc. (Swiftwater, PA)
Comparison of lymphocytic immunoreactivity and histopathological scores
Two general approaches can be used to evaluate the severity of cellular inflammatory infiltration in the CNS induced by viruses with different neurovirulence. The conventional semi-quantitative approach relies on histopathological analysis of routinely stained (H&E and/or Nissl) sections and assignment of cellular inflammatory infiltration (CII) scores based on the number of perivascular infiltrates and degree of parenchymal infiltration. The representative H&E-stained sections that contained
Acknowledgements
We acknowledge the staff of Bioqual, Inc. (Rockville, MD) and Pathology Associates Division of Charles River Laboratories (Frederick, MD), and Dr. J. Ward, Dr. R. Montali, Lawrence Faucette, Marina Rahman, and Katherine Shea (Comparative Medicine Branch, NIAID, NIH) for their help in conducting the studies. We also thank Drs. J. Taubenberger and S. Whitehead, NIAID, NIH for helpful discussions and critical reading of the manuscript. This work was supported by funds provided by the NIAID
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