Protocol for potent activation of T cells using BPI-stimulated murine bone marrow-derived cells

In vitro T cell activation and differentiation are fundamental techniques in immunological research. Here, we present a protocol for the potent activation of T cells using bactericidal/permeability-increasing protein (BPI)-stimulated murine bone marrow-derived cells (BMDCs). We describe steps for preparing supernatants harvested from BPI-stimulated BMDCs (SN BPI). We then detail procedures for purification of (naive) CD4⁺ T cells from murine spleens and peripheral lymph nodes and their consecutive polyclonal activation in the presence of SN BPI. For complete details on the use and execution of this protocol, please refer to Bülow et al.¹.