Allergic Peritonitis (AP)
8-9-week-old C57BL/6 (WT) females and/or males were sensitized on days
-14 and -7 with subcutaneous (s.c) injection of 100μg of OVA (A5378-10G,
Sigma-Aldrich, St. Louis, MO,
USA) + 1μg of SEB (S4881-5mg, Sigma-Aldrich) in 200μl of PBS (#1927718,
BI, Beit-Haemek, IL) per mouse (OVA [500μg/ml]/SEB [5 μg/ml])
and challenged at day 0 with i.p injection of 10μg of OVA + 0.1μg SEB in
200μl of PBS per mouse (OVA [50µg/ml]/SEB [0.5 µg/ml]) or PBS
only. Mice were euthanized by CO2 inhalation on days 0
(0h), 2 (48h), 4 (96h), 7(168h) and 10 (240h) and peritoneal lavage was
performed with 3 ml PBSX1+3% FCS. Peritoneal cells were counted in
hemocytometer by Trypan blue (#03-102-1B, BI) staining. Peritoneal
lavages were centrifuged at 150g, 5min, 4oC and
supernatants were collected and stored or immediately used for further
analysis. Cell pellets were resuspended in Flow Cytometry (FC) buffer
(0.5% BSA + 2% FCS in PBSX1) for FC analysis.