BMMCs sensitization with serum from immune OVA/SEB mice and activation with either OVA or SEB
BMMCs were prepared as previously described (16). Thereafter, BMMCs were sensitized o.n with 1:5 diluted serum harvested from OVA/SEB challenged mice 48h after the challenge. The day after sensitized BMMCs were washed and resuspended in BMMCs culture medium and activated for 24h at 37oC with either 0.5µg/ml OVA or 10µg/ml SEB. After incubation supernatants were collected and analyzed for TNFα levels.
Sash mice intraperitoneal reconstitution with bone marrow derived MCs (BMMCs)
OVA/SEB AP model was induced in 6-12-week-old female Sash mice as described above. Twenty-four hours before challenge with OVA/SEB, mice were reconstituted i.p with 2×106 BMMCs/200μl PBS obtained from C57BL/6. Mice were euthanized 48h after challenge and peritoneal lavages (3ml) were processed as described above.