Imaging and analysis on the interaction between human antigen-pulsed V82 T cells and antigen-specific CD4 T cells

This protocol describes how to visualize surface protein-protein co-localization across a cell-cell interface between antigen-presenting y8-T cells and CD4 T cells. By consolidating immunofluorescence assay, confocal microscopy and 3D imaging analysis, it enables assessment of interaction between cell surface proteins such as A42PD1 and TLR4 between co-cultured y8-T and CD4 T cells. This protocol can be applied to study a surface protein of interest and its poten-tial interaction with a target cell/protein at the cell-cell interface. For complete details on the use and execution of this profile, please refer to Mo et al. (2020).