Determination of interferon-gamma in peripheral blood mononuclear cells treated with aqueous extract of gynura procumbens / Putri Amirah Azman

Gynura procumbens (Lour.), locally known as "sambung nyawa" is annual evergreen shrub that can be found in Southeast Asia. G.procumbens is used to treat many ailments such as inflammation. G.procumbens'' anti-inflammatory and immunomodulatory activity may be utilized for treatment of inflammatory disease. The immune system play crucial role in in the pathogenesis of inflammatory disorders which can be treated using drug that have anti-inflammatory and immunostimulant properties. Interferon-gamma (IFN-y) is type II interferon family, secreted by activated T cell and Natural Killer (NK) cells plays role in macrophage activation, inflammation, T helper (Th 1 cell responses) and immunoediting. Cytokine such as Interferon-y is detected using peripheral blood mononuclear cells (PBMC) because PBMC contain lymphocytes that consist of T cell, B cell, NK cells, monocytes and dendritic cells (Miyahira, 2012). Although G.procumbens has known to have anti-inflammatory properties, the expression of Interferon-gamma towards G.procumbens have yet to be fully discovered. Therefore, we would like to determine the expression of Interferon-y in PBMC treated with aqueous extract of G.procumbens using Luminex Human Magnetic Assay. In the present study, we first demonstrated the phytochemical properties of G.procumbens. Then, 12 ml of whole blood is separated using Ficoll density gradient centrifugation to obtain Peripheral blood mononuclear cells (PBMC). PBMC is treated with G.procumbens with addition of CLI 095 and Polymyxin B as inhibitor. The expression of Interferon-y is measured using Luminex assay. The results show that the expression of Interferon-y in PBMC treated with aqueous extract of G.procumbens showed extremely low value in three different concentration and test group. The results for all groups showed extremely low value (<49.259 I>g/ml) of expression towards Interferon-y where statistically analysis were not necessary. In conclusion, we demonstrate the Peripheral Blood Mononuclear Cells (PBMC) treated with aqueous extract of Gprocmbens do not possess the expression of Interferon-gamma (IFN-y) via TLR 4 receptors and MyD88 pathway.