Fig. 1

GLS-1027 inhibits pro-inflammatory cytokine release in human PBMC in vitro. (A) GLS-1027 exhibits cell-type-specific inhibition of LPS-induced TNF-α production. TNF-α levels were measured 48 h post stimulation with LPS (5 µg/ml) in absence or presence of GLS-1027 (10 µM). (B) GLS-1027 does not inhibit CD3/CD28 mediated T cell activation. Purified human CD3⁺ T cells were stimulated with anti-CD3/CD28 beads (10 µg/ml) or ConA in the presence or absence of GLS-1027 (10 µM), and IFN-γ production was measured via ELISpot immune assay. (C) T cell proliferation in the presence or absence of 10 µM GLS-1027 was analyzed by ³H‐thymidine uptake lymphoproliferative assay after co-stimulation through CD3.⁺CD28 using plate bound antibodies. (D-E) GLS-1027 exhibits dose-dependent inhibition of TNF-α and IL-1β secretion from LPS-stimulated cells. PBMC were isolated and cultured in the presence or absence of LPS 5ug/ml with or without GLS-1027 (0.1,1.0 or 10 µM). Cytokine levels (pg/ml) in cell-free supernatants were assayed by ELISA as described under Materials and Methods. Error bars in all panels indicate 1 standard deviation among 3 replicates per condition. The experiment was repeated twice with similar results are shown in this representative experiment