TCR transgenic clone selection guided by immune receptor analysis and single-cell RNA expression of polyclonal responders
- Laboratory of Immunoregulation and Mucosal Immunology, VIB Center for Inflammation Research, Belgium
- Department of Internal Medicine and Pediatrics, Ghent University, Belgium
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmaceutical Sciences, Ghent University, Belgium
- VIB Single Cell Core, VIB Center, Belgium
- Department of Applied Mathematics, Computer Science and Statistics, Ghent University, Belgium
- Department of Head and Skin, Ghent University, Belgium
- Department of Pulmonary Medicine, Erasmus University Medical Center Rotterdam, Netherlands
Figures
Figure 1 with 1 supplement
Schematic overview of the generation of rationalized CORSET8 transgenic mice.
(I) Selection of the immunogenic epitope: here based on T cell epitope mapping results of Zhuang et al., 2021, which were confirmed by Erez et al., 2023 (Erez et al., 2023; Zhuang et al., 2021). (II)…
Figure 1—figure supplement 1
Screening of CD8 T cell hybridoma clones and validation of first generation CORSET8 mice.
(A) Mean fluorescence intensity (MFI) of IL-2 for individual CD8 T cell hybridoma clones. Based on the fold increase of IL-2 between medium control and Spike peptide pool stimulation, clone 47 was …
Figure 2 with 1 supplement
Combined single-cell and T cell receptor (TCR) analysis using DALI identified the most promising T cell clone.
(A) Uniform manifold approximation and projection (UMAP) of splenic CD8 T cell single-cell RNA sequencing visualizing four different clusters based on single-cell analysis. (B) Projection of the …
Figure 2—figure supplement 1
Gating strategy used to sort Spike Tetramer positive and Tetramer negative splenic CD8 T cells.
Flow cytometry plots from a naive and a representative immunized mouse are shown. In total, three samples were subjected for single-cell analysis: (1) Tetramer positive, CD44+ cells pooled from two …
Figure 3
CORSET8 mice exhibit normal T cell populations and T cell receptor (TCR) Tg cells recognize target peptide ex vivo.
(A) Proportion of CD4 T and CD8 T cells among CD3+ TCRβ+ T cells in the spleen of CORSET8 mice and wildtype C57BL/6 littermates. (B) Proportion of CD62L+ CD44−, CD62L+ CD44+, and CD62− CD44+ T cells …
Figure 4
CORSET8 cells proliferate in vivo in response to Spike immunization or SARS-CoV-2 infection.
(A, B) Schematic setup of the experiments. (C) CTV dilution profile 4 days after intratracheal immunization with Spike/CpG or intramuscular immunization with Pfizer BNT162b2 vaccine. Quantification …
Author response image 1
