Oral Presentation 23rd Annual Lorne Proteomics Symposium 2018

Temporal quantitative proteomics of human CD4+ and CD8+ T-cell activation using tandem mass spectrometry and SWATH (#4)

Harshi Weerakoon 1 2 , Jeremy Potriquet 3 , Yide Wong 1 3 , Oscar Haigh 4 , Ailin Lepletier 1 , John J Miles 1 3 , Jason Mulvenna 2 , Michelle M Hill 1 5
  1. QIMR Berghofer Medical Reserach Institution, Herston, QLD, Australia
  2. School of Biomedical Sciences, The University of Queensland, St Lucia, QLD, Australia
  3. Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, QLD, Australia
  4. Translational Research Institute, Woolloongabba, QLD, Australia
  5. The University of Queensland Diamantina Institute, The University of Queensland, Woolloongabba, QLD, Australia

Aberrant T-cell activation or termination is a characteristic of numerous inflammatory and auto-immune diseases. Furthermore, dysfunction of anti-tumour T-cells (‘exhaustion’) has emerged as an important aspect of cancer development. While some of the signalling events and transcription factors involved in T-cell activation are well-established, a systems map of human T-cell activation circuits remains to be generated. In particular, there are limited quantitative proteomics studies on various human T-cell lineages which have distinct functions. To understand proteomic changes associated with different T-cell lineages, we use a relatively new mass spectrometry-based label free quantification method; SWATH (sequential window acquisition of all theoretical fragment masses) to characterize the temporal changes in activated CD4+ and CD8+ T-cell proteomes. A consensus spectral library was generated from a set of shotgun T-cell proteomics data with fractionation at cellular, protein and peptide levels. SWATH experiments were then performed at 8 time points following in-vitro T-cell receptor activation over 7 days, on both CD4+ and CD8+ T-cells subsets from 3 volunteers. A total of 2,100 proteins were quantified and ~500 of these proteins were significantly changed over the course of T-cell activation and termination. Major changes to both CD4+ and CD8+ T-cell proteomic profiles occurred 24 hours after activation, peaking on the 3rd day. Functional annotation of these significantly changed proteins revealed activation phase-specific modulations of energy metabolism pathways up to Day 3. Thereafter, upregulation of cell death and apoptosis pathways, and downregulation of energy metabolism pathways were revealed as possible regulatory networks associated with termination of T-cell activation. This study generated the first temporal quantitative proteome of human CD4+ and CD8+ T-cell activation, providing a baseline for future studies.