For generating mixed BMCs, 8- to 10-week-old GF Jh−/− mice were transferred to an autoclaved plastic cylinder sealed with Mylar film. The cylinder-containing GF mice were irradiated with a lethal dose of 900 cGy using X-RAD 320 (Precision X-ray). Irradiated recipient mice were transferred to the isolator and reconstituted with 5 × 106 mixed BM cells. BM cells from Jh−/− mice were mixed with BM cells from either Icosl−/− or H2-Ab1−/− mice in an 8:2 ratio. An 8:2 mixture of Jh−/− BM cells and WT B6 BM cells was transferred into irradiated GF Jh−/− mice as a control. At 6 weeks after reconstitution, BMCs were analyzed. For elucidating the role of long-lived IgE-producing cells in sustained IgE responses, 14-week-old GF B6 mice were similarly irradiated with 900 cGy and reconstituted with BM cells from Rag1−/− mice to deplete lymphocytes.

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