Fig 2: A phenotypically CD8 positive subset of human B cells.Mobilized apheresis material was analyzed with multicolor flow cytometry. The CD8+ TCR negative population was gated (A), and the cells included in the thick black gate resolved using the markers CD335 for NK cells and CD19 for B cells (B). Percentage of both populations is given in the quadrants, and black arrows point to the CD8 expression profiles and MFI values of the boxed CD335+ NK cells and CD19+ B cells, respectively. One representative donor out of four shown. (C) TCR-negative cells were flow sorted into CD8 high, CD8 medium, and CD8 dim populations along with TCR-CD8- cells as negative controls, as indicated by the dashed boxes on panel (A). Expression of genes in the sorted population was then measured by quantitative real-time PCR, and is shown as percentage of the GAPDH housekeeping gene (* for NCR, encoding CD335, the scale on the right hand side applies). One representative donor out of two shown.

Fig 3: Expansion and tissue infiltration of cytotoxic T subsets in IgG4-RD.(A) UMAP plot of subgroups of T cells. CTL, cytotoxic T lymphocyte; Treg, regulatory T cell; MAIT, mucosal-associated invariant T cell; DN_T, double negative T cell; T/Mono, doublets of T cells and monocytes. (B) Violin plot showing expression distribution of canonical cell markers in cell types shown in A. (C) Comparison of proportions of each cell type between the 2 groups (7 HCs, 9 IgG4-RD patients). Results are shown as mean ± SD. Exact 2-tailed P values are given. Significance was determined by 2-tailed Student’s t test (CD4+ naive T, CD4+ effector memory T, CD4+ memory T, CD4+ CTL, Treg, IFN-stimulated CD4+ T, CD8+ CTL-GZMK, CD8+ CTL-GNLY, and DN_T) or Mann-Whitney test (CD8+ naive T, MAIT, γδT, and prolif. lymph). (D) Representative tissue multicolor immunofluorescence images and modeling analyses (repeated 3 times for each tissue type). Original magnification, ×200. (E) Comparisons of infiltrated T cell subsets in different tissues (lymph node n = 9, salivary gland n = 5, pancreas n = 13, lacrimal gland n = 10). Kruskal-Wallis test followed by post hoc Dunn’s multiple-comparison test was performed. (F) Volcano plot showing the differentially expressed genes between T cells from IgG4-RD and HCs. (G) GO analysis using Metascape for genes that were upregulated in T cells from IgG4-RD. (H) The expression levels of well-defined chemotaxis- and cytotoxicity-related genes. (I) The chemotaxis score, cytotoxicity score, and score of inhibitory receptors in T cell subpopulations. Significance determined by Mann-Whitney test. (J) Constituent ratios of follicular helper T (Tfh) cells and type 2 Tfh (Tfh2) cells among CD4+ T cells (3 HCs, 5 IgG4-RD patients). Significance determined by χ2 test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Fig 4: TIGIT is elevated on CD8+T cell in patients with GC and significantly reduced after D2 gastrectomy. (a) Flow cytometry analysis of CD8+ TIGIT+, CD8+PD-1+, or CD8+ TIGIT+PD-1+ T-cell populations in pre- and post-operative PBMC from patients with GC. (b) Frequency and number of CD8+ TIGIT+, CD8+PD-1+, or CD8+ TIGIT+PD-1+ cells in pre-chemotherapy and post-chemotherapy treated PBMC from patients with GC. **P < 0.01.

Fig 5: Combination of SOX and anti-TIGIT is beneficial to CD8+ T cells proliferation and IFN-γ release. (a, b) CD8+ TIL cells were treated with SOX, Anti-TIGIT, or SOX +Anti-TIGIT. Immunofluorescence was used to detect expression levels of KI-67. (c, d) CD8+ TIL cells were treated with SOX, Anti-TIGIT, or SOX +Anti-TIGIT, and stimulated with PMA for 5 h. Frequency and number of IFN-γ of CD8+ cells were quantified. *P < 0.05, **P < 0.01.