The aim of this study was to investigate the effect of IL13 and CLA on differentiation properties of Macrophages to M2 type using an acute monocytic leukemia (THP1) and further measurement of CD206 (MR) and CD163 as a markers of differentiation using FACS analyzer. Also, to investigate whether M2 differentiation is up regulated through the activation of PPARγ, by the addition of its antagonist GW9662 to down regulate CD206 (MR) expression. The human monocytic cell line THP-1 cell was first derived from the peripheral blood of a 1 year old male with acute monocytic leukemia. Cell Culture Manipulation and sub-culture for the expression CD206 (MR) in CLA and IL-13-treated THP-1 cells. THEN CD206 (MR) expression in CLA and IL-13-treated THP-1 cells in the presence of GW9662. In this study the In-vitro differentiation of monocytes to M2 were enhanced by using Conjugated linoleic acid (CLA) isomers and IL-13, The result shows significant increase in MR (M2 marker) expression on the surface of differentiated cells as well there was significant decrease (P>.005) in the expression of CD 163 suggesting no up regulation occur. The expression of M2 markers (CD206) and PPARγ, a nuclear receptor controlling macrophage inflammation, correlate positively. This was clear when cells were treated with the PPARγ antagonist GW9662 (1μM), the amount of CD206 expression was reduced in both (IL13 and CLA). In conclusion the conducted study shows that CLA and IL-13 enhance the ability of Macrophages to differentiate from M1 to M2 thus promote the anti-inflammatory M2 macrophages phenotype also it demonstrated that PPARγ activation induces polarization of monocytes towards a beneficial M2 phenotype suggesting that CLA and IL-13 partly modulate their action through PPARγ activation.
Read full abstract