252-LB: Oxidative Stress Induces Toll-Like Receptor -2 and -4 in Human Peripheral Blood Mononuclear Cells: Implications for Metabolic Inflammation

Background: Toll-like receptors (TLRs) are emerging as immunometabolic receptors and nutrient sensors. In obese and T2D patients, oxidative stress acts as a critical early-trigger of altered pathophysiology. In immune cells, it remains unclear whether the oxidative stress can modulate expression of the receptors. We assessed the expression of TLR2 and TLR4 in the peripheral blood mononuclear cells (PBMCs) following H 2 O 2 treatment. Methods: Human PBMCs were isolated from healthy donors and were subjected to oxidative stress by H 2 O 2 with or without antioxidants or ROS inhibitors. Using qRT-PCR and flow cytometry, the gene expression of TLR2 and TLR4 was determined, Moreover, expression of the phosphorylated c-Jun, ERK1/2, p38, and NF-?B signaling proteins was determined. Results: TLR2 and TLR4 gene and protein expression was elevated in PBMCs following induction of oxidative stress compared to controls (P 2 O 2 -mediated TLR2 and TLR4 upregulation. Oxidative stress also increased the gene expression of proinflammatory IL-1β, IL-6, TNF-α, IFN-γ and MCP-1 (P Conclusion: Taken together, oxidative stress upregulates the TLR2 and TLR4 expression in PBMC through the MAPK/NF-?B dependent mechanism in parallel with increased expression of proinflammatory cytokines/chemokine. We speculate that TLR2/TLR4 suppression by ROS-inhibition or antioxidant strategies may alleviate metabolic inflammation. Disclosure S.T. Sindhu: None. A. Al Madhoun: None. N. Akhter: None. F. Al-Mulla: None. R. Ahmad: None. Funding Kuwait Foundation for the Advancement of Sciences; Dasman Diabetes Institute