Figure legends
Figure 1. Higher activation status of CD4+ T cells from primary biliary cholangitis (PBC) patients. (a ) Comparison of the proportions of CD4+ T cells in total T cells in PBC (n=42) and health control (HC) (n=20) with representative plots of flow cytometry for each group in the left panel and scatter-plot in the right panel. (b ) Comparison of the proportions of naïve CD4+ T cells in total CD4+T cells in PBC and HC groups with representative plots of flow cytometry for each group in the left panel and scatter-plot in the right panel. (c ) In PBC, the ratios of activated (HLA-DR+/ICOS+) CD4+ T cells were higher than that in HC.
Figure 2. Increased multiple-cytokine activation of CD4+ T Cells from patients with PBC. (a ) Comparison of proportions of Th1/Th2/Th17 (IFN-γ+/IL-4+/IL-17+CD4+ T cells) in total CD4+ T cells in PBC and HC groups with representative plots of flow cytometry for each group in the left panel and scatter-plot in the right panel.
Figure 3. Increased expression of Lysosomal associated membrane protein 2 isoform A (LAMP-2A) in PBC naïve CD4+ T Cells, and its relationship with disease severity. Mean fluorescence intensity (MFI) of LAMP-2A in activated (a ) (HLA-DR+), (b ) (ICOS+) CD4+ T cells were significantly higher than non-activated (HLA-DR/ICOS) ones, either in PBC or HC group. (c ) LAMP-2A expressions in non-activated (HLA-DR/ICOS) CD4+ T cells from PBC patients were greater than that in HC. (d ) Higher LAMP-2A expressions in naïve CD4+ T cells from PBC patients than that in HC. (e-f ) LAMP-2A expressions in naïve CD4+ T cells from PBC patients were independent of gender, age, the presence of anti-mitochondrial antibody (AMA), and a variety of clinical biochemical indicators reflecting liver function (ALT, AST, ALP, GGT, TBil and Alb ). (g ) (h ) LAMP-2A expressions in naïve CD4+ T cells from PBC patients with advanced clinical stage were significantly increased, in term of pathological stage as well as the degree of liver fibrosis.
Figure 4. Naïve CD4+ T cells from PBC patients were more liable to be activated than HC counterparts, and this abnormal hyperactivity could be reversed by deleting the gene encoding LAMP-2A. (a ) Comparison of the proliferation ability of naïve CD4+ T cells in PBC, HC groups and PBC naïve CD4+ T cells transfected with LAMP-2A-specific shRNA (PBC-L2A) after stimulation by CD3/CD28 activator in vitro. Representative histograms of flow cytometry for each group were in the left panel and column chart in the right panel. (b ) Comparison of the apoptosis of naïve CD4+ T cells in PBC, HC and PBC-L2A groups, with representative histograms of flow cytometry for each group in the left panel and column chart in the right panel. (c ) IFN-γ and IL-2 levels in the cell supernatant of naïve CD4+ T cells in PBC, HC and PBC-L2A groups were quantified by enzyme-linked immunosorbent assay (ELISA), including resting naïve CD4+ T cells (Rest) and cells following activation by CD3/CD28 activator (Act). * p < 0.05; *** P < 0.001.