Immunization of Guinea pigs with Adeno-vectors
Animals.
Hartley outbread female guinea pigs (Harlan)
Age: 6 weeks
Overview groups.
Group 1:
Group 2:
Group 3:
Group 4:
Ad6tetOE1F78E2p7 (T212)
Ad6F78E2662 (T212)
Ad6HVR1E2662 (T212)
Ad6F78E2684 (N2)
Animal identification.
All animals can be identified by the small holes in their ear. The group number of each animal can
be devised from the amount of holes in the animal’s left ear, while the identification number within
the group can be devised from the amount of holes in the animal’s right ear.
Group 1:
Group 2:
Group 3:
Group 4:
6 animals
6 animals
5 animals
4 animals
(1.0 to 1.5)
(2.0 to 2.5)
(3.0 to 3.4)
(4.0 to 4.3)
Technical information.
All animals will be anesthetized before bleeding using a cocktail of ketamine (Ketalar; 6,75 mg/kg
body weight) and xylazine (Rompun; 5 mg/kg). The anesthetic will be injected intramuscularly.
The following volumes will be administered per 100 g of body weight.
Ketalar (15 mg/ml): 45 l
Rompun (2%, 20 mg/ml): 25 l
At the indicated time points, serum will be collected via cardiac puncture. Animals will be
anesthetized and blood will be collected using a 23G needle. Blood will be allowed to cloth for at
least 30 minutes at room temperature. Serum will be collected after centrifugation (5 minutes @
4.500xg). and stored at -80°C.
At the indicated time points, all animals will be vaccinated with 1010 adenoviral particles prepared
in 100 l buffer A195. Prepare fresh vaccine solution for each vaccination point according to the
attached dilution scheme. Prepare and keep all vaccination solution at 4°C just until
administration.
Each dose will be injected intramuscularly using a 1 ml syringe with a 25G needle. The dose (100
l) will be divided over the two rear legs of the animals and injected in the quadriceps muscle (50
l in each leg). The remainder of the vaccines will be refrozen and kept at -80°C, but may not be
used for a subsequent vaccination point.
Schedule.
Time point
Date
Manipulation
Sample
preparation
Day 0
04 June (groups 1&2)
05 June (groups 3&4)
Bleeding
Vaccination #1
Serum
Week 4
29 June (all animals)
Bleeding (start 10h00)
Serum
Vaccination #2
Immediate shipment samples
collected on day 0 and week 4.
Week 6
13 July (all animals)
Bleeding (start 10h00)
Immediate shipment samples
Serum
17 July
Optional sacrifice
serum
Week 12
24-27 August
Optional bleeding
Optional vaccination #3
Serum
Week 14
7-11 September
Optional sacrifice
Serum
Recombinant E1E2 vaccination.
Animals.
Hartley outbread female guinea pigs (Harlan)
Age: 6 weeks
Overview groups.
Group 5:
recombinant E1E2
Animal identification.
All animals can be identified by the small holes in their ear. The group number of each animal can
be devised from the amount of holes in the animal’s left ear, while the identification number within
the group can be devised from the amount of holes in the animal’s right ear.
Group 5:
6 animals
(5.0 to 5.5)
Technical information.
All animals will be anesthetized before bleeding using a cocktail of ketamine (Ketalar; 6,75 mg/kg
body weight) and xylazine (Rompun; 5 mg/kg). The anesthetic will be injected intramuscularly.
The following volumes will be administered per 100 g of body weight.
Ketalar (15 mg/ml): 45 l
Rompun (2%, 20 mg/ml): 25 l
At the indicated time points, serum will be collected via cardiac puncture. Animals will be
anesthetized and blood will be collected using a 23G needle. Blood will be allowed to cloth for at
least 30 minutes at room temperature. Serum will be collected after centrifugation (5 minutes @
4.500xg). and stored at -80°C.
At the indicated time points, all animals will be vaccinated with …..
Schedule.
Time point
Date
Manipulation
Sample
preparation
Day 0
17 July (group 5)
Bleeding
Vaccination #1
Serum
Day 31
17 August (group 5)
Vaccination #2
Day 90
15 October
Vaccination #3
Day 104
29 October
Bleeding and sacrifice
serum