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.