title: Infection- and procedure-dependent effects on pulmonary gene expression in the early phase of influenza A virus infection in mice.
abstract: BACKGROUND
  Investigating the host response in the early stage of influenza A virus (IAV) infection is of considerable interest. However, it is conceivable that effects due to the anesthesia and/or intranasal infection procedure might introduce artifacts. We therefore aimed to evaluate the effects of anesthesia and/or intranasal infection on transcription of selected pulmonary mRNAs in two inbred mouse strains with differential susceptibility to IAV infection.
  
  
  RESULTS
  DBA/2J and C57BL/6J mice were evaluated in a time course experiment in which lung tissue was sampled after 6, 12, 18, 24, 48 and 120 h. After anesthesia with ketamine and xylazine, a suspension of mouse-adapted IAV strain PR8_Mun in 20 μl sterile buffer, or 20 μl sterile buffer only, was instilled intranasally. The mice receiving anesthesia and PBS only were designated the "mock treatment" group. Pulmonary expression of 10 host mRNAs (Fos, Retnla, Irg1, Il6, Il1b, Cxcl10, Stat1, Ifng, Ifnl2, and Mx1) and viral hemagglutinin (HA) mRNA were determined at the designated time points. As expected, weight loss and viral replication were greater in the DBA/2J strain (which is more susceptible to IAV infection). Four mRNAs (Retnla, Irg1, Il6, and Cxcl10) were procedure-dependently regulated in DBA/2J mice between 6 and 24 h, and two (Retnla and Il6) in C57BL/6J mice, although to a lesser extent. All 10 mRNAs rose after infection, but one (Fos) only in DBA/2J mice. These infection-dependent effects could be separated from procedure-dependent effects beginning around 12 h in DBA/2J and 18 h in C57BL/6J mice. The interferon-related mRNAs Stat1, Ifng, Infl2, and Mx1 were unaffected by mock treatment in either mouse strain. Mx1 and Infl2 correlated best with HA mRNA expression (r = 0.97 and 0.93, respectively, in DBA/2J).
  
  
  CONCLUSIONS
  These results demonstrate effects of the anesthesia and/or intranasal infection procedure on pulmonary gene expression, which are detectable between approximately 6 and 24 h post procedure and vary in intensity and temporal evolution depending on the mouse strain used. Mock infection controls should be included in all studies on pulmonary gene expression in the early phase of infection with IAV and, likely, other respiratory pathogens.
authors.forename: Matthias
authors.initials: M
authors.lastname: Preusse
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Tantawy
authors.forename: Frank
authors.initials: F
authors.lastname: Klawonn
authors.forename: Klaus
authors.initials: K
authors.lastname: Schughart
authors.affiliation: Institute for Experimental Infection Research, TWINCORE Center for Experimental and Clinical Infection Research, Feodor-Lynen-Str, 7, Hannover 30625, Germany. Frank.Pessler@twincore.de.
authors.forename: Frank
authors.initials: F
authors.lastname: Pessler
mesh_terms: D000818:Animals; D001835:Body Weight; D004195:Disease Models, Animal; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D054884:Host-Pathogen Interactions; D009980:Influenza A virus / Q000502:physiology*; D008168:Lung / Q000473:pathology* / Q000821:virology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D009976:Orthomyxoviridae Infections / Q000473:pathology* / Q000821:virology*; D013997:Time Factors; D019562:Viral Load