Mice Lacking Expression of Secondary Lymphoid Organ Chemokine Have Defects in Lymphocyte Homing and Dendritic Cell Localization

Gunn, Michael D.; Kyuwa, Shigeru; Tam, Carmen; Kakiuchi, Terutaka; Matsuzawa, Akio; Williams, Lewis T.; Nakano, Hideki

Contact Sensitization. The shaved abdomens of mice were painted with 0.4 ml of 5 mg/ml FITC dissolved in a 50:50 (vol/ vol) mixture of acetone and dibutylphthalate. After 24 h, cell suspensions of pooled inguinal, axillary, and brachial LNs were prepared from each mouse and analyzed by flow cytometry. In some experiments, aliquots of LN cells were stained with biotinylated anti-CD11c followed by SA-PerCP. To determine I-A expression on FITC Ï© cells, aliquots of LN cells from four mice were pooled, partially purified on metrizamide, stained with biotinylated anti-I-Ad followed by SA-PerCP, and analyzed by flow cytometry. For sensitization with a lower dose of antigen, 25 l of 8 mg/ml FITC was spotted on the shaved right flank of five Ï© / Ï© and five plt mice. After 24 h, the draining and contralateral inguinal LNs were removed and analyzed individually. Single cell suspensions were quantitated, stained with anti-I-Ad and anti-B220, and analyzed by flow cytometry.