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Updated: Apr 5, 2026
Isolation and Th17 Differentiation of Naïve CD4 T Lymphocytes
Published on: September 26, 2013
The Advanced Lipoxidation end Product Precursor Malondialdehyde Induces IL-17E Expression and Skews Lymphocytes to
Abstract:
Malondialdehyde (MDA) is a highly reactive endogenous product of thromboxane synthesis in the prostagland and lipid peroxidation by reactive oxygen species. Elevated MDA levels occur in diabetes and atherosclerotic plaques. The aim of this study was to examine the molecular mechanisms of MDA-induced IL-17E cytokine expression and its effect on T-cell differentiation. Real-time PCR, RT-PCR and ELISA were used to assess the expression of IL-17 family cytokines in Jurkat T-cells and human peripheral blood lymphocytes (PBLCs) from diabetic subjects. Luciferase reporter assays were used for the promoter activation study. Pharmacological inhibitors were used for signaling pathway experiments. FACS analyses were used to measure the Th1, Th2 and Th17 subset levels. MDA induced significant (2- to 3-fold; p < 0.01) generation of IL-17E mRNA in a dose- and time-dependent manner in Jurkat T-cells and PBLCs. Elevated IL-17E mRNA levels were found in the lymphocytes from diabetic subjects. The increased IL-17E protein and mRNA levels correlate well with serum MDA levels from diabetic patients. Transient transfection of plasmid containing the minimum IL-17E promoter region (pIL-17E-Luc) showed a significant (2-fold; p < 0.01) increase in luciferase activity. Pretreatment of lymphocytes with pharmacological inhibitors showed the involvement of antioxidant, NF-ƙB, p38MAPK, PKC and ERK signaling pathways. Quantification of the Th1, Th2 and Th17 cell population in PBLCs via FACS analyses revealed an increase in the Th17 subset. These results show that MDA transcriptionally upregulates the expression of IL-17E in lymphocytes and alters lymphocyte differentiation towards the pathogenic Th17 subset.
Insights
Malondialdehyde (MDA) significantly increases Interleukin-17E (IL-17E) cytokine expression in T-cells. This MDA-induced IL-17E promotes T-cell differentiation towards the pathogenic Th17 subset, particularly in diabetes.
Area of Science:
- Immunology
- Molecular Biology
- Endocrinology
Background:
- Malondialdehyde (MDA) is a reactive product of lipid peroxidation, with elevated levels linked to diabetes and atherosclerosis.
- Interleukin-17E (IL-17E) is a cytokine implicated in inflammatory processes.
Purpose of the Study:
- To investigate the molecular mechanisms by which MDA induces IL-17E expression.
- To determine the impact of MDA on T-cell differentiation, focusing on Th17 subset development.
Main Methods:
- Real-time PCR, RT-PCR, and ELISA were employed to quantify IL-17 family cytokine expression in T-cells and lymphocytes.
- Luciferase reporter assays assessed IL-17E promoter activity.
- Pharmacological inhibitors and FACS analysis were used to investigate signaling pathways and T-cell subset populations (Th1, Th2, Th17).
Main Results:
- MDA dose- and time-dependently upregulated IL-17E mRNA in Jurkat T-cells and human peripheral blood lymphocytes (PBLCs).
- Elevated IL-17E mRNA and protein levels correlated with serum MDA in diabetic subjects.
- MDA-induced IL-17E expression involved NF-κB, p38MAPK, PKC, and ERK signaling pathways.
- FACS analysis revealed an expansion of the Th17 cell subset.
Conclusions:
- MDA transcriptionally upregulates IL-17E expression in lymphocytes.
- MDA promotes T-cell differentiation towards the pathogenic Th17 subset, suggesting a role in diabetes-related inflammation.
