Figure 2. mAb CD69.2.2 induces CD69 internalization in activated lymphocytes in vitro and inhibition of CD69 expression in vivo. (A) CD69+ lymphocytes, previously stained with mAb CD69.2.2 antibodies and GAMalexa488, were incubated at 4°C or 37°C for 1 hour (left column). Incubation with PBS (pH 2.0; right column), removes membrane-bound mAbs. Representative photographs were taken by confocal microscopy. (B-C) C57BL/6 mice were treated at 8 weeks of age with a single injection (500 μg intraperitoneally) of mAb CD69.2.2 or control mAb (IgG1). Lymphocytes from treated mice were analyzed by flow cytometry. Representative profiles of B cells (B220/CD69) in lymph node (LN), spleen, and peritoneum 15 days after injection are shown (B). Bone marrow and LN lymphocytes (C) were subjected to flow cytometric analysis. Representative profiles of B220/IgM, IgD/IgM, LY6G/B220, and CD3/B220 are shown with the percentage of cells in each quadrant.
Figure 2.

mAb CD69.2.2 induces CD69 internalization in activated lymphocytes in vitro and inhibition of CD69 expression in vivo. (A) CD69+ lymphocytes, previously stained with mAb CD69.2.2 antibodies and GAMalexa488, were incubated at 4°C or 37°C for 1 hour (left column). Incubation with PBS (pH 2.0; right column), removes membrane-bound mAbs. Representative photographs were taken by confocal microscopy. (B-C) C57BL/6 mice were treated at 8 weeks of age with a single injection (500 μg intraperitoneally) of mAb CD69.2.2 or control mAb (IgG1). Lymphocytes from treated mice were analyzed by flow cytometry. Representative profiles of B cells (B220/CD69) in lymph node (LN), spleen, and peritoneum 15 days after injection are shown (B). Bone marrow and LN lymphocytes (C) were subjected to flow cytometric analysis. Representative profiles of B220/IgM, IgD/IgM, LY6G/B220, and CD3/B220 are shown with the percentage of cells in each quadrant.

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