Schughart2 project protocol

Susceptibility to influenza A (H1N1) viral infection in females of 53 BXD recombinant inbred strains of mice   (2012)

Schughart K, Williams RW
With: Nedelko T, Kollmus H, Klawonn F, Spijker S, Lu L, Heßman M, Alberts R




Schughart2_Protocol

Project protocol — Contents
Workflow
Equipment, supplies
Reagents, solutions
Procedure: Influenza A (H1N1) infection and host response
Definitions
Data
References

Workflow

Step
Procedure
Influenza A virus PR8 (H1N1)
Equipment
Data Collected
1
H1N1 virus cultured and infectivity titrated
-
incubator
-
2
Mice weighed and anesthetized

-
balance scale
baseline body weight
3
 Mice infected intranasally
2 x 103 FFU
nebulizer (1 mL syringe)
-
4
 Body weight and survival monitored for 13 days
-
balance scale
daily body weights, survival

See Abbreviations

Equipment, supplies

  • Balance scale
  • Syringes, needles

Reagents, solutions

  • Influenza A/Puerto Rico/8/34 (H1N1; PR8M) adapted for mouse; virus propagated in 10d embryonated chicken eggs.
  • Ketamine-Xylazine in sterile NaCl (50 mg/mL Ketamine (Invesa Arzneimittel GmbH, Freiburg Germany); 2% Xylazine (Bayer Health-Care, Leverkusen)
  • Phosphate buffered saline (PBS)

Procedure: Influenza A virus PR8 (H1N1) infection and host response

a. Mice are weighed prior to infection (baseline).
b. Mice are anesthetized by intra-peritoneal injection of Ketamine-Xylazine combination at a dose based on individual body weight.
c. H1N1 virus is administered intranasally (2 x 103 FFU in 20 µL sterile PBS).
d. Health is monitored over a period of 13 days by assessing degree of weight loss, general health, and survival of infected mice.
e. Dead mice are noted; moribund mice or mice that have lost >25% of their body weight are euthanized and recorded as dead. (It should be noted that for mice that did not show any signs of body weight loss over the entire time period after infection, additional parameters were not obtained to verify that they were infected.)
f. Strains are designated susceptible (=0) or resistant (=1), with susceptible characterized by rapid weight loss (>25%) and death (Note: designations for susceptible (=0) or resistant (=1) for each strain are assigned to individual mice in the MPD dataset).

Definitions

Abbreviations (see also MPD units and abbreviations)
Influenza A virus PR8
FFU
 Focus forming units; a measure of virus titer in an infected lung

Data collected by investigator

  • percent change in body weight
  • survival (time to death in days)
  • susceptibility to viral infection