Fig 1: TIGIT+ cells display a distinct cytokine profile. (a) Representative dot plots showing intracellular cytokine production after cultivating CLL PBMCs for 24 h with CD3/CD28 activating beads. (b) Cytokine production of TIGIT- or TIGIT+CD4+ T cells in 14 samples. (c) Mean fluorescence intensity ratio (MFIR) of CD155 and CD112 on CD5+CD19+ CLL (top) or CD5+ T cells (bottom). The histograms show representative FACS plots of CLL cells (gated for CD5+CD19+cells) and T cells (CD5+ cells) stained with isotype controls (in gray) and CD112/CD155 specific antibodies (in black). The dot plots show results from n = 14 samples.
Fig 2: Expression levels of TIGIT and PD-1 in TIGIT+ PD-1+ CXCR5- CD4+ T, TIGIT- PD-1+ CXCR5- CD4+ T and TIGIT+ PD-1- CXCR5- CD4+ T cells in patients with RA and the HC. (A) Percentage of TIGIT+ PD-1+ CXCR5- CD4+ T cells, (B) TIGIT- PD-1+ CXCR5- CD4+ T cells, (C) TIGIT- PD-1+ CXCR5- CD4+ T cells, the MFI of (D) TIGIT and (E) PD-1 in TIGIT+ PD-1+ CXCR5- CD4+ T cells, (F) the MFI of PD-1 in TIGIT- PD-1+ CXCR5- CD4+ T cells, (G) the MFI of TIGIT in TIGIT+ PD-1- CXCR5- CD4+ T cells between patients with RA and the HC. (H) The MFI of TIGIT in TIGIT+ PD-1+ CXCR5- CD4+ T cells and TIGIT+ PD-1- CXCR5- CD4+ T cells from patients with RA. (I) The MFI of PD-1 in TIGIT+ PD-1+ CXCR5- CD4+ T cells and TIGIT-PD-1+CXCR5-CD4+T cells from patients with RA. HC, healthy controls; MFI, mean fluorescence intensity; PD-1, programmed cell death protein 1; RA, rheumatoid arthritis; TIGIT, T cell immunoreceptor with immunoglobulin and ITIM domain; Ig, immunoglobulin; CXCR5, C-X-C chemokine receptor type 5.
Fig 3: T cells from CLL patients display elevated TIGIT expression. (a-b) Peripheral blood samples from CLL patients (CLL) and age-matched healthy controls (HC) were analyzed by flow cytometry (FACS) with regard to TIGIT, 2B4 and PD-1 expression on CD4 or CD8 T cells. (c) Distribution of inhibitory receptors on TIGIT- or TIGIT+ T cells. (d) Correlation of TIGIT and PD-1 expression on CD4+ or CD8+ T cells. (e) Distribution of CD4+TIGIT+ cells in patients divided according to clinical markers of disease burden (Rai/ Binet stage or treatment status).
Fig 4: TIGIT is particularly expressed on antigen experienced T cells. (a) T cell subsets in peripheral blood samples from CLL patients and age-matched healthy controls (HC) were measured by FACS analysis defined by CD62 L and CD45RA. Plots represent naïve (Tnaive: CD62 L+CD45RA+), central memory (TCM: CD62 L+CD45RA-), effector memory (TEM: CD62 L-CD45RA-) and terminally differentiated effector memory (TEMRA: CD62 L-CD45RA+). (b) T cell subset distribution in the TIGIT- and TIGIT+ T cell compartment. (c) Absolute cell counts (cells/µL blood) of CD4+ and CD8+ subsets expressing TIGIT.
Fig 5: Absolute cell counts of TIGIT+Th1, TIGIT+Treg and TIGIT+Tfh cells are significantly increased in CLL. (a) Plot of percentages of Th1, Th2, Treg and Tfh among CD4+ T cells from controls and CLL patients. (b) Percentage of TIGIT expressing Th1, Th2, Treg and Tfh cells. (c) Absolute cell counts (cells/µL). (d) Percentage of CD226+ cells among TIGIT- versus TIGIT+ CD4+ or CD8+ cells in HCs or CLL patients.(e) Correlation of TIGIT and CD226 expression on CD4+ or CD8+ T cells.
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