Schughart1 project protocol

Susceptibility to influenza A (H1N1) viral infection in females of 9 strains of mice   (2009)

Schughart K
With: Srivastava B, Błazejewska P, Hessmann M, Bruder D, Geffers R, Mauel S, Gruber AD




Schughart1_Protocol

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

Workflow

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

-
balance scale
baseline body weight
3
 Mice infected intranasally
2 x 103 FFU
nebulizer (1 mL syringe)
-
4
 Weight loss and survival monitored for 14 days
-
balance scale
daily body weights, survival

See Abbreviations

Equipment, supplies

  • Balance scale
  • Syringes, 1 mL

Reagents, solutions

  • Influenza A/Puerto Rico/8/34 (H1N1; PR8) adapted for mouse; virus propagated in the chorioallantoic cavity of 10d embryonated hen eggs for 48h at 37°C, and titrated according to Srivastava et al., 2009.
  • Ketamine-Xylazine (Rompun®) combination anesthetic

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 14 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.
f. Strains are designated susceptible (=0) or resistant (=1), with susceptible characterized by rapid weight loss (>25%) and death within seven days of infection (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
PBS
 Phosphate buffered saline

Data collected by investigator

• percent change in body weight

• susceptibility to viral infection