Balb/c Wt in vivo treatment with OVA / Vitamin D3.
Female Balb/c wild-type mice at the age of 8-9 weeks were treated at day
0 intranasally with 25μl PBS, PBS + Vitamin D3 (=1α,25
DihodroxyvitaminD3 from Sigma Aldrich, 10 ng), Ovalbumin (500 μg) or OVA
+ Vitamin D3 (500μg and 10 ng respectively, with 20 min time delay in
between) according to their group. After 48h the treatment was repeated.
After additional 48 h whole lung cells were isolated as described
previously and cultured with anti-CD3 (10μg/ml) respectively
anti-CD3(10μg/ml) and anti-CD28 (1μg/ml) antibodies17.