(P14-010-24) Development of a Model to Investigate Diet-induced Inflammation Using Peripheral Blood Mononuclear Cells (PBMC)

Objectives: Develop a model to investigate diet-induced inflammation using Peripheral Blood Mononuclear Cells (PBMC).

Methods: PBMC were collected before and after (4 h) from individuals fed a High Fat Moderate Carbohydrate meal (HFMC) to investigate the NFκB signaling pathway using an immunofluorescence method. In a separate experiment, fasting human whole blood samples treated with lipopolysaccharide (LPS, 1ng/mL) for 4 h at 37° C with and without a phosphatase inhibitor (Halt™) was used to understand inflammatory signaling. Interleukin-6 (IL-6) was measured as a marker of inflammation using the ELISA method before and after LPS+/-Halt™ treatments.

Results: PBMC collected from individuals fed with HFMC indicated activation of the NFκB signaling pathway as observed by translocation of P50 Subunits from cytoplasm to nucleus at 4 h after HFMC meal ingestion. PBMCs collected after incubation of fasting whole blood with LPS (4 h) had a similar presentation as observed in the 4 h post meal PBMCs. There are several limitations in quantifying the inflammatory status in PBMC florescent images as it was a translocation of NFκB subunits from the cytoplasm to the nucleus. However, fasting human whole blood samples treated with LPS for 4 h showed significantly increased IL-6 concentration compared to the baseline (0 h, pre-treatment, 2.1 ± 0.5 Vs. 1.4 ± 0.5 ng/mL, P< 0.05). LPS-induced IL-6 production was inhibited by Halt™ (2.1 ± 0.5 Vs. 1.2 ± 0.4, P< 0.05), possibly due to inhibition of phosphorylation (i.e. NFκB) and preventing downstream inflammatory signaling pathway.

Conclusions: The data from this study help us to further our knowledge in the development of an in-vitro model to explore diet-induced inflammation (i.e., postprandial inflammation) using PBMC.

Funding Sources: N/A