Independently of CD146, the sSS patients exhibited increased CD31 expression on CD4 and CD8 cells; some showed loss of CD28 from CD4 cells (Supporting information, Fig. S8). Other memory,

adhesion and homing markers were similar to those in HDs. Thus, circulating T cells in the few CTD patients who exhibited phenotypic T cell activation had increased CD146 expression, associated with a broadened range of activation markers. We examined CD146 expression on circulating CD4 and CD8 T cells of HDs and patients with CTDs, and characterized the relationship of Selleck Cobimetinib CD146 with surface markers associated with activation, memory, adhesion and homing. As expected, CD146 expression correlated with some activation and memory markers, but unexpected differences between CD4 and CD8 T cells were observed. CD146 on T cells was increased in a small number of patients with sSS, all of whom exhibited systemic T cell activation, but not in patients with other CTDs, who did not. Previous work has shown CD146 induction by phytohaemagglutinin-activated T cells [3, 7]. We found that stimulation of HD T cells with anti-CD3/anti-CD28, a more physiological stimulus, up-regulated CD146 expression with slower kinetics and longer persistence than CD69, but similar to CD25. Both activated CD4 and CD8 T cells expressed

CD146. Ex vivo, however, the relationship of CD146 expression to T cell activation was more complex. Selleck BGB324 CD146-expressing CD4 T cells contained a greater proportion of activated-phenotype cells than bulk CD4 Cell press T cells (OX40+, CD69+ and low-level

CD25 expression). Within the CD4 subset, the CD146+ population comprised almost exclusively CD45RO+/RA–/CD28+ non-senescent memory cells, and was enriched in CD27− cells, suggesting repeated activation. Nevertheless, the correlation with activation was not absolute: most activated cells lacked CD146, and no single marker correlated perfectly with CD146 expression. Thus, CD4 T cell activation in vivo does not induce CD146 expression as uniformly as it does in vitro. This could partly reflect differences in the timing of expression of activation markers post-stimulation but suggests that physiological stimuli induce CD146 expression more selectively than is recapitulated in vitro. A few CD146+CD4 T cells are FoxP3+ CD25high, consistent with a Treg phenotype, but FoxP3 can be expressed by human activated effector T cells and additional markers would be required to address this definitively [33]. Previous work has reported similar findings, albeit with fewer markers analysed in individual donors [7]. Unexpectedly, the association of CD146 with activation and memory ex vivo was less marked in CD8 T cells. In HD CD8 cells, CD146-expressing cells were less frequent than in CD4 cells; of the activation markers studied, only CD69 was enriched significantly in CD146+ CD8 cells.