CD3+ B-1a Cells as a Mediator of Disease Progression in Autoimmune-Prone Mice

Figure 6

Intracellular staining for the detection of IFNγ, TNFα, and IL-10. Mononuclear cells isolated from the peripheral blood were cultured with LPS (10 μg/ml), PMA (50 ng/ml), ionomycin (500 ng/ml), and monensin (2 μM) for 5 h. After culture, the cells were stained with appropriate fluorescence antibodies to detect cell-surface markers, fixed, and permeabilized. The cells were also stained intracellularly with APC-conjugated anti-IFNγ, anti-TNFα, and anti-IL-10. After washing, the cells were immediately subjected to flow cytometric analysis. (a) Representative results of the flow cytometry of the B6/lpr mice showing intracellular staining of IFNγ (left column), TNFα (middle column), and IL-10 (right column) of B cells (upper line), T cells (middle line), and CD3+B220+ cells (lower line). (b) Representative results of flow cytometry of the MRL/lpr mice showing intracellular staining of IFNγ (left column), TNFα (middle column), and IL-10 (right column) of B cells (upper line), T cells (middle line), and CD3+B220+ cells (lower line).

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