Project
protocol
-
Contents
Workflow
and
sampling
Step |
Procedure |
Data
collected |
1 |
Mouse-adapted H3N2 produced and prepared for intra-nasal delivery |
- |
2 |
Mice weighed to determine subsequent weight changes |
Body weight |
3 |
Mice anesthetized |
- |
4 |
Mice infected with H3N2 intra-nasally or with PBS alone |
- |
5 |
Body weight determined daily post-infection |
Body weight |
6 |
Mice euthanized and blood samples taken for hematology analysis |
Leukocyte profiles |
7 |
Lungs harvested and weighed |
Lung weight |
Equipment
and
supplies
- Balance
- Syringes and needles
- Micro tubes containing EDTA (Sarstedt)
- VetScan HM5 (Abaxis)
Reagents
and
solutions
- Ketamine/xylazine (100 mg/mL and 20 mg/mL, respectively)
- H3N2: mouse-adapted virus strain influenza A/HK/01/68 obtained from Otto Haller, University of Freiburg GERMANY
- Natrium chloride solution
- Phosphate-buffered saline
- Isofluroran (cp-pharma, Burgdorf GERMANY)
Procedure: H3N2 preparation/delivery, daily body weights and terminal leukocyte profiles and lung weight
- H3N2 is produced in the allantoic cavity of 10-d-old embryonated hen eggs for 48h at 37°C.
- Mice are weighed to determine dosing.
- Mice are anesthetized by intra-peritoneal injection with a mixture of Ketamine/Xylazine in sterile natrium chloride solution.
- Mice are intra-nasally infected with 20 µL virus solution of sterile phospate-buffered saline (PBS) using a concentration of 10 focus-forming units (FFU), or with PBS alone.
- Body weight is measured daily and determined as percentage of initial weight at day 0 (pre-infection).
- Mice are euthanized with Isofluoran (160 µL) and exsanguinated via puncture of the retro-bulbar vein plexus and lungs are collected.
- Blood is immediately measured in a hematologic system (VetScan).
- Lungs are weighed.
- Relative lung weight is calculated as: (lung weight on day of preparation / body weight on day of preparation) x 100.
Data
collected
by
investigator
- Body weight pre-infection
- Body weight post-infection (expressed as percentage of pre-infection weight)
- Leukocyte profiles
- Lung weight
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