Enrichment of RAW264.7 macrophages with essential 18-carbon fatty acids affects both respiratory burst and production of immune modulating cytokines

Abstract

Macrophages play a vital role in the innate immune system. Thereby, production of both reactive oxygen intermediates and immune modulating cytokines is crucial for successful pathogen defense. Fatty acids may interfere with immune response in several ways. In this study, we investigated the influence of essential polyunsaturated fatty acids (PUFA) on key macrophage functions. RAW264.7 macrophages were cultured in a medium supplemented with 2 or 15 μmol/L of the n-6 PUFA linoleic acid (LA) or of the n-3 PUFA α-linolenic acid (LNA), respectively. Cells were tested for incorporation of fatty acids as well as NADPH oxidase activity. Furthermore, supernatants were collected for detection of NO and cytokine release (TNF-α, IL-6, IL-10). Exposure of RAW264.7 macrophages to LA or LNA resulted in incorporation of these fatty acids and their derivatives. Thereby, supplementation with both LA and LNA caused a significant increase in NADPH oxidase activity. In contrast, synthesis of NO was not affected by PUFA supplementation. Moreover, distinct effects could be seen in the release of immune modulating cytokines. Due to enhancement of NADPH oxidase activity, PUFA presumably promote the killing of pathogens crucial in host defense. In addition, the unsaturated fatty acids tested in our study were shown to modulate cytokine release by the macrophages, thus driving immune response into an anti-inflammatory direction. Of note, distinct differences between the n-6 PUFA LA and the n-3 PUFA LNA underline the impact of PUFA family on immune response.