Metabolomic Profiling of the Immune Stimulatory Effect of Eicosenoids on PMA-Differentiated THP-1 Cells

Abstract

Honey bee venom has been established to have significant effect in immunotherapy. In the present study, (Z)-11-eicosenol-a major constituent of bee venom, along with its derivations methyl cis-11-eicosenoate and cis-11-eicosenoic acid, were synthesised to investigate their immune stimulatory effect and possible use as vaccine adjuvants. Stimuli that prime and activate the immune system have exerted profound effects on immune cells, particularly macrophages; however, the effectiveness of bee venom constituents as immune stimulants has not yet been established. Here, the abilities of these compounds to act as pro-inflammatory stimuli were assessed, either alone or in combination with lipopolysaccharide (LPS), by examining the secretion of tumour necrosis factor-α (TNF-α) and the cytokines interleukin-1β (IL-1β), IL-6 and IL-10 by THP-1 macrophages. The compounds clearly increased the levels of IL-1β and decreased IL-10, whereas a decrease in IL-6 levels suggested a complex mechanism of action. A more in-depth profile of macrophage behaviour was therefore obtained by comprehensive untargeted metabolic profiling of the cells using liquid chromatography mass spectrometry (LC-MS) to confirm the ability of the eicosanoids to trigger the immune system. The level of 358 polar and 315 non-polar metabolites were changed significantly (p < 0.05) by all treatments. The LPS-stimulated production of most of the inflammatory metabolite biomarkers in glycolysis, the tricarboxylic acid (TCA) cycle, the pentose phosphate pathway, purine, pyrimidine and fatty acids metabolism were significantly enhanced by all three compounds, and particularly by methyl cis-11-eicosenoate and cis-11-eicosenoic acid. These findings support the proposed actions of (Z)-11-eicosenol, methyl cis-11-eicosenoate and cis-11-eicosenoic acid as immune system stimulators.

Pharmacological effects and mechanisms of bee venom and its main components: Recent progress and perspective

“…Similarly, the same research group also studied the metabolomic profiling for the immune stimulatory effect of a major constituent of BV, (Z)-11-eicosenol, and its derivations methyl cis-11-eicosenoate and cis-11-eicosenoic acid on PMA-differentiated THP-1 cells. The results supported the proposed actions of the three components as immune system stimulators ( Alqarni et al, 2019 ). Besides, bvPLA 2 has attenuated several immune system-related diseases.…”

Toxicological and bioactivity evaluation of blackcurrant press cake, sea buckthorn leaves and bark from Scots pine and Norway spruce extracts under a green integrated approach

“…THP-1 cells (2 × 10 5 /mL) were differentiated for 48 h using a 50 ng/ mL solution of PMA as described in Alqarni et al (2019). After the 24 h incubation in PMA free medium, the extracts at different concentrations (7-1667 μg/mL) were added to cells and thereafter LPS (0.5 μg/mL) was added.…”

“…Effects of extracts on THP-1 promonocyte viability was measured as described in Alqarni et al (2019). Briefly, the THP-1 cells were added into 96-well plates at a density of 1 × 10 5 cells per well and incubated for 24 h in an incubator (at 37 • C, in 5% CO 2 and 100% RH).…”

Lack of Nck1 protein and Nck-CD3 interaction caused the increment of lipid content in Jurkat T cells

“…Methyl cis-11-eicosenoate, or eicosenoic acid (cis-11), is formed by the elongation of oleic acid, which is then converted to docosenoic acid and nervonic acid. In the human monocyte cell line (THP-1), eicosenoic acid (cis-11) promotes IL-1β and IL-6 production, but inhibits the production of IL-10 [ 34 ]. Thus, decreased eicosenoic acid (cis-11) in Nck1 deficient cells might affect cytokine production by T cells.…”