Open Access

Antonius A. de Waard, Tamara Verkerk, Marlieke L. M. Jongsma, Kelly Hoefakker, Sunesh Sethumadhavan, Carolin Gerke, Sophie Bliss, George M. C. Janssen, Arnoud H. de Ru, Frans H. J. Claas, Arend Mulder, Robert Tampé, Peter A. van Veelen, Anne Halenius, Robbert M. Spaapen

• With the emergence of immunotherapies, the understanding of functional HLA class I antigen presentation to T cells is more relevant than ever. Current knowledge on antigen presentation is based on decades of research in a wide variety of cell types with varying antigen presentation machinery (APM) expression patterns, proteomes and HLA haplotypes. This diversity complicates the establishment of individual APM contributions to antigen generation, selection and presentation. Therefore, we generated a novel Panel of APM Knockout Cell lines (PAKC) from the same genetic origin. After CRISPR/Cas9 genome-editing of ten individual APM components in a human cell line, we derived clonal cell lines and confirmed their knockout status and phenotype. We then show how PAKC will accelerate research on the functional interplay between APM components and their role in antigen generation and presentation. This will lead to improved understanding of peptide-specific T cell responses in infection, cancer and autoimmunity.