Eosinophils and respiratory virus infection: a dual-standard curve qRT-PCR-based method for determining virus recovery from mouse lung tissue

Methods Mol Biol. 2014:1178:257-66. doi: 10.1007/978-1-4939-1016-8_22.

Abstract

Several lines of investigation have indicated a role for eosinophilic leukocytes in limiting virus infectivity and promoting virion clearance. We have established a respiratory virus infection model with pneumonia virus of mice (PVM; family Paramyxoviridae), a natural mouse pathogen that replicates the more severe forms of human disease elicited by the phylogenetically related respiratory syncytial virus (RSV). In this chapter, we present a rapid and highly reproducible dual-standard curve qRT-PCR based method for quantitative detection of PVM replication in mouse lung tissue. We have used this assay to evaluate eosinophil-mediated antiviral host defense in mouse models of cytokine and antigen-driven eosinophilic inflammation.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Eosinophils / cytology*
  • Eosinophils / physiology
  • Lung / immunology*
  • Lung / virology*
  • Mice
  • Polymerase Chain Reaction
  • Respiratory Syncytial Viruses / immunology
  • Respiratory Syncytial Viruses / isolation & purification*