Transcriptional reprogramming via signaling domains of CD2, CD28 and 4-1BB

The goal of the project was to assess transcriptomic changes induced by costimulatory signals.A reporter line based on the human Jurkat T cell line E6.1 was engineered to express chimeric receptors harboring signaling domains derived from human CD2, CD28 and 4-1BB fused to the extracellular domain of mouse ICOS. In addition, reporter cells expressing mouse ICOS lacking a cytoplasmic domain was generated. The resulting reporter cells were stimulated with control T cell stimulator cells (TCS-control) that engaged the TCR/CD3 complex alone or with TCS that co-engaged the TCR/CD3 complex and the respective chimeric mICOS receptor (TCS-mICOSL). TCS are based on the murine thymoma line BW5147 and express a membrane bound anti-CD3 single chain antibody. Following 24 h of co-culture the cells were harvested und subjected to RNA-seq analysis.