NGS data produced in 'Rapid selection and identification of functional CD8+ T-cell epitopes from large peptide-coding libraries'; Nature Communications (2019)

Sharma, G et al. Rapid selection and identification of functional CD8+ T-cell epitopes from large peptide-coding libraries. Nature Communications. Accepted (August 2019)

Abstract:

Cytotoxic CD8+ T-cells recognize and eliminate infected or malignant cells that present, at their cell surfaces, short peptide epitopes derived from intracellularly processed antigens. However, broadly searching for specific major histocompatibility complex (MHC)-bound peptide epitopes that are naturally processed and capable of eliciting a functional T-cell response has been challenging. Here, we report a method for deep and unbiased T-cell epitope profiling, which is done by using in vitro co-culture of CD8+ T-cells and target cells transduced with high-complexity epitope-encoding minigene libraries. Target cells that are subject to cytotoxic attack from T-cells in co-culture are isolated, before they are lost to apoptosis, by fluorescence-activated cell sorting and characterized by sequencing the minigenes encoded within. In the present study, we validate this highly parallelized method using known murine T-cell receptor/peptide-MHC pairs and diverse minigene-encoded epitope libraries to identify naturally processed and MHC-presented peptide epitopes unambiguously and with high sensitivity.