Ethyl Caffeate (ETHYL 3,4-DIHYDROXYCINNAMATE) suppressed the differentiation of naive CD4+ T cells into Th1 in vitro. Furthermore, Ethyl Caffeate intensely blocked the transcriptional expression in interferon-γ-related signaling, including IFN-γ, T-bet, STAT1, and STAT4.

In vivo, ETHYL CAFFEATE(ECF) treatment reduced the severity of collagen-induced arthritis (CIA), inhibited IFN-γ and IL-6 secretion, and?decreased the proportion of CD11b+Gr-1+ splenic neutrophil.?Meanwhile, ECF treatment significantly inhibited the IFN-γ expression in CD4+T cell without obviously influencing the development of Th17 cells and T regulatory cells.

Shikui X , Aixue Z , Zengjun G , et al. Ethyl Caffeate Ameliorates Collagen-Induced Arthritis by Suppressing Th1 Immune Response[J]. Journal of Immunology Research, 2017, 2017:1-11.

The proliferation of splenocytes or T cells in response to ConA, LPS, and anti-CD3/28 was measured by CCK-8 Kit.?Briefly, BALB/c splenocyte suspension (5?×?105 cells/well) was cultured with ConA (5?μg/ml), LPS (10?μg/ml), and anti-CD3 (5?μg/ml;?145-2C11) in the presence of ECF at indicated concentrations.?The cultures were incubated for 48?h, 20?μl of CCK-8 was then added to each well before the end of culture, and OD value was read at 450?nm.?The MTT method was used to measure the cytotoxicity of the sample.?Splenocytes (5?×?105 cells/well) were cultured in triplicates in the absence or presence of ECF in a 96-well flat-bottomed plate (Costar) for 48?h.?MTT (5?mg/ml) was pulsed for 4?h prior to the end of the culture.?Upon removal of MTT/medium, 150?μl of DMSO was added to each well, and the plate was agitated on an oscillator for 5?min to dissolve the precipitate.?The assay plate was read at 570?nm using a microplate reader.

Collagen was dissolved in 0.1 M acetic acid at 4°C overnight.?Male DBA/1 mice were immunized at the tail base with 125?μg of collagen emulsified in complete Freund's adjuvant (CFA) containing Mycobacterium tuberculosis strain H37Rv.?Each mouse was then boosted with the same amount of collagen plus CFA 21 days later (taken as day 0). Starting from day 9 for 10 consecutive days, the mice were administered daily with ECF (50?mg/kg) or methotrexate (2?mg/kg)