Table 3.
Cell proliferative assay and cytokine production in the splenocyte cultures obtained from immunized mice
|
Immunization route
a
|
Immunization regimen |
Cytokine production (pg/ml)
b
|
||||
|---|---|---|---|---|---|---|
| IL-2 | IFN-γ | IL-4 | IL-5 | SI c | ||
| Intramuscular vaccination |
Saline |
12 ± 3 |
13 ± 8 |
<10 |
<10 |
0.23 |
| |
pVAX1 |
16 ± 4 |
15 ± 5 |
11 ± 8 |
<10 |
0.51 |
| |
pVAX1-MEG-CTXA2/B |
93 ± 21 |
385 ± 64 |
15 ± 7 |
13 ± 6 |
1.61 |
| Intranasal vaccination |
BRD509 |
13 ± 4 |
14 ± 5 |
<10 |
17 ± 5 |
0.75 |
| |
BRD509/pVAX1 |
14 ± 7 |
15 ± 6 |
<10 |
19 ± 5 |
1.07 |
| |
BRD509/ pVAX1-MEG-CTXA2/B |
105 ± 30 |
412 ± 24 |
23 ± 6 |
<10 |
2.12 |
| Intraoral vaccination |
BRD509 |
12 ± 5 |
15 ± 4 |
<10 |
16 ± 8 |
0.75 |
| |
BRD509/pVAX1 |
15 ± 7 |
18 ± 4 |
<10 |
16 ± 7 |
1.45 |
| BRD509/ pVAX1-MEG-CTXA2/B | 121 ± 36 | 507 ± 16 | <10 | 12 ± 4 | 2.85 | |
aMice were immunized by three immunization routes, intramuscular, intraoral, intranasal on day 0 and day 14 and day 28 with different immunization regimens.
bThe splenocyte culture supernatants taken from mice (n = 3, each group) 2 weeks after the last immunization were examined for cytokine production by sandwich ELISA obtained at 24 h for IL-4, at 36 h for IL-5, at 72 h for IL-2 and 96 h for IFN-γ.
cThe results of proliferation assays are expressed as the stimulation index (SI), calculated as the ratio between the mean counts per minute (cpm) for triplicate stimulated cultures and the mean counts per min for triplicate unstimulated cultures. SI values 2.5-fold greater than the SI of the control groups were considered as significant.