Epitope Mapping of Anti-Diacylglycerol Kinase ζ Monoclonal Antibody for the Detection of T Cells by Immunohistochemical Analyses

The diacylglycerol kinases (DGKs) are a family of proteins that catalyze the phosphorylation of the cell membrane lipid diacylglycerol (DG), a cellular component that is important in lipid biochemistry and signal transduction, into phosphatidic acid. DG-mediated signal transduction downstream of the T cell receptor has previously been reported to be terminated in most cases by one of 10 DGK isoforms, DGKζ. In this study, we performed immunohistochemical analysis using a rabbit anti-DGKζ monoclonal antibody (mAb) (clone EPR22040-80) against tissues from the tonsils of a patient with oropharyngeal squamous cell carcinoma. We demonstrated that many DGKζ-expressing T cells are localized in the tonsils. We further characterized the binding epitope using an enzyme-linked immunosorbent assay and found that Pro790, Gln791, Gly792, and Leu795 residues of DGKζ are important for facilitating anti-DGKζ mAb binding to DGKζ. This anti-DGKζ mAb could be valuable in immunohistochemical analyses in determining the distribution of DGKζ-expressing T cells in pathophysiological tissues.