Imaging antigen-induced PI3K activation in T cells.

Activation of phosphoinositide 3-kinase (PI3K) at the immunological synapse between a T cell and an antigen-presenting cell (APC) has not been demonstrated. Using fluorescent-specific probes, we show here that the formation of an immunological synapse led to sustained production of 3'-phosphoinositides in the T cell, whereby phosphatidylinositol-3,4,5-trisphosphate (PIP3) but not phosphatidylinositol-3,4-bisphosphate was localized to the cell membrane. The accumulation of PIP3 after T cell activation preceded the increase in intracellular calcium. Neither the formation of conjugates between T cells and APCs nor signaling events such as phosphotyrosine accumulation and calcium increase changed substantially when PI3K was inhibited, and only a limited reduction in synthesis of interleukin 2 occurred. In T cell-APC conjugates, PIP3 accumulated at the T cell-APC synapse as well as in the rest of the T cell plasma membrane, which indicated unusual regulation of PI3K activity during antigen presentation.