Large Scale HLA Tetramer Tracking of T Cells Throughout Dengue Infection Using Mass Cytometry

T cells play important multi-faceted roles during dengue infection and understanding their responses is important for defining correlates of protective immunity and identifying effective vaccine antigens. Using mass cytometry and a highly-multiplexed peptide-human leukocyte antigen (HLA) tetramer staining strategy, we probed T cells from dengue patients for a total of 430 dengue and control candidate epitopes together with key markers of activation, trafficking, and differentiation.
Conclusion:
During acute disease, dengue-specific CD8+ T cells expressed a distinct profile of activation and trafficking receptors that distinguished them from non-dengue-specific T cells. During convalescence, dengue-specific T cells differentiated into two major cell fates, CD57+ CD127- terminally differentiated senescent memory cells and CD127+ CD57- proliferation-capable memory cells.
Notes:
Preprocessing: After mass cytometry acquisition, the signal of each parameter was normalized based on the equilibration beads added to each sample (Finck et al., 2013). All zero values were randomized by an R-script that uniformly distributes values between minus-one and zero. Individual samples were debarcoded manually in FlowJo v10 (Treestar, Inc.). Gated for DNA_1 and DNA_2 double positive, Cisplatin-negative, CD14-negative, CD45-positive cells. Experiments 1 and 2 have a different staining panel than Experiment 3. Experiments 1 and 2 were acquired on the CyTOF 2 while Experiment 3 was acquired on the Helios. The same control sample FMFG was used in all experiments.