Heterogeneity of circulating CD8 T-cells specific to islet, neo-antigen and virus in patients with type 1 diabetes mellitus
Sandra Laban (Leiden University Medical Center)
Jessica S. Suwandi (Leiden University Medical Center)
Vincent van Unen (Leiden University Medical Center)
Jos Pool (Leiden University Medical Center)
Joris Wesselius (Leiden University Medical Center)
Thomas Hollt (TU Delft - Computer Graphics and Visualisation, Leiden University Medical Center)
Nicola Pezzotti (TU Delft - Computer Graphics and Visualisation)
Anna Vilanova (TU Delft - Computer Graphics and Visualisation)
Boudewijn P.F. Lelieveldt (Leiden University Medical Center)
Bart O. Roep (Leiden University Medical Center, City of Hope National Medical Center)
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Abstract
Auto-reactive CD8 T-cells play an important role in the destruction of pancreatic β-cells resulting in type 1 diabetes (T1D). However, the phenotype of these auto-reactive cytolytic CD8 T-cells has not yet been extensively described. We used high-dimensional mass cytometry to phenotype autoantigen- (pre-proinsulin), neoantigen- (insulin-DRIP) and virus-(cytomegalovirus) reactive CD8 T-cells in peripheral blood mononuclear cells (PBMCs) of T1D patients. A panel of 33 monoclonal antibodies was designed to further characterise these cells at the single-cell level. HLA-A2 class I tetramers were used for the detection of antigen-specific CD8 T-cells. Using a novel Hierarchical Stochastic Neighbor Embedding (HSNE) tool (implemented in Cytosplore), we identified 42 clusters within the CD8 T-cell compartment of three T1D patients and revealed profound heterogeneity between individuals, as each patient displayed a distinct cluster distribution. Single-cell analysis of pre-proinsulin, insulin-DRIP and cytomegalovirus-specific CD8 T-cells showed that the detected specificities were heterogeneous between and within patients. These findings emphasize the challenge to define the obscure nature of auto-reactive CD8 T-cells.
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