Figure 5.

Expression of immunomodulatory molecules in cultured human LNSCs. (A) Induction of transforming growth factor beta 1 (TGFB1) CD40, CD80, CD274 (PD-L1; programmed cell death 1 ligand), nitric oxide synthase 2 (NOS2), and indoleamine 2,3-dioxygenase 1 (IDO1) was assessed by qPCR in cultured LNSCs (passages 4–9) after stimulation with IFNγ at different time points (4, 24, and 48 h). Data are represented as fold induction (median with interquartile range) by comparing the mRNA levels in stimulated cells to corresponding unstimulated cells in 15 donors (n = 5 per donor group). The dotted line represents a fold induction of 1. (B) CD40 protein expression was measured by flow cytometry in CD45− stromal cells. Histogram depicts the increase in staining between isotype, unstimulated and stimulated for 72 h with IFNγ and is displayed for one representative donor (RA-risk individuals passage 8) out of 13 donors tested. (C) Induction of CD40 protein upon stimulation with IFNγ for 72 h is further presented as Mean Fluorescent Intensity (MFI) ratio (isotype/staining) in all donors measured (healthy individuals n = 5, RA-risk individuals n = 5, and RA patients n = 3, passages 4–9).