Poster Presentation International Veterinary Immunology Symposium 2016

T cell receptor repertoire analysis using next-generation sequencing. (#143)

Timothy Connelley 1 , Alie Gillespie 2 , Deepali Vasoya 1 , Andy Law 1 , Mick Watson 1 , Cynthia Baldwin 2 , Ivan Morrison 1
  1. The Roslin Institute, Roslin, MIDLOTHIAN, United Kingdom
  2. University of Massachussetts, Amherst, Connecticut, USA

T-cell recognition of antigens is mediated through the somatically rearranged, clonotypically distributed, heterodimeric T-cell receptor (TCR). The TCR repertoire is hugely diverse, providing a challenge to its comprehensive characterisation. TCR chain analysis by Sanger sequencing is limited by the number of sequences that can be feasibly generated and so provides comparatively low resolution data on TCR repertoires. The advent of NGS technologies has provided a step-change in the level of detailed information that can be generated on TCR repertoires. We have employed a SMART-based unbiased PCR system, the Illumina MiSeq platform and a modified version of the MiTCR bioinformatic pipeline to analyse bovine TCR repertoires. The system uses a set of generic technologies that should be easily transferable to other veterinary species and affords the opportunity to rapidly enhance immunological analyses of T-cell responses.