recombinant IL-10 reduces the production of TNF-α from whole-blood cultures from healthy individuals stimulated with LPS. This anti-inflammatory effect of IL-10 is not reproduced in whole-blood cultures from individuals with diabetes.1 (link) Therefore, we tested whether THP-1 macrophages exposed to high levels of glucose developed this resistance to IL-10 (via TNF-α reduction). The cells were plated for 1 hour, as mentioned previously (on 24-well plates, 37°C, 5% CO2), and then stimulated with 5 µg/mL LPS in the presence or absence of recombinant IL-10 (10 ng/mL; R&D systems, Minneapolis, MN, USA). Supernatants were collected after 6 hours of incubation and frozen at −80°C to be further studied for TNF-α cytokine concentration, as described elsewhere.1 (link)
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Glucose Impairs IL-10 Anti-Inflammatory Effect
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Other organizations : Presbyterian College, Wake Forest University
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Variable analysis
independent variables
- Presence or absence of recombinant IL-10 (10 ng/mL)
- TNF-α cytokine concentration in the supernatant
- THP-1 macrophages
- LPS concentration (5 µg/mL)
- Incubation time (6 hours)
- Cell culture conditions (24-well plates, 37°C, 5% CO2)
- Positive control: THP-1 macrophages stimulated with LPS
- Negative control: Not explicitly mentioned
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