Article Figures & Data
Figures
- FIGURE 1.
Polymyxin B, tyrothricin, gramicidin, and neomycin trigger IL-1β secretion. A, Murine BMDCs were primed with LPS and exposed to members of five different classes of antibiotics at a concentration of 50 μg/ml, and IL-1β secretion was measured in supernatants after 6 h of stimulation. ATP was used as a positive control. Note that only polymyxin B, tyrothricin, gramicidin, and neomycin induced IL-1β secretion. B, LPS primed BMDCs from TNFR1/2 and IL-1R1–deficient mice were exposed to 50 μg/ml of polymyxin B, tyrothricin, gramicidin, and neomycin. IL-1β secretion was measured in supernatants after 6 h of stimulation. Data are expressed as the mean ± SD from three independent experiments, all performed in triplicates.
- FIGURE 2.
IL-1β secretion triggered by polymyxin B, tyrothricin, neomycin, and gramicidin requires all components of the NLRP3 inflammasome. A, Wild-type and NLRP3-deficient mice derived BMDCs were primed with LPS and exposed 50 μg/ml of polymyxin B, tyrothricin, gramicidin, and neomycin. IL-1β secretion was measured in supernatants after 6 h of stimulation. ATP was used as a positive control. B, Immunoblot analysis of mature IL-1β (p17) and caspase-1 cleavage product (p10) in wild-type and NLRP3-deficient BMDCs stimulated with antibiotics. C, IL-1β ELISA for wild-type and ASC-deficient BMDCs stimulated with antibiotics. Note that IL-1β secretion was dependent on the presence of NLRP3 and ASC for all drugs. D, Wild-type BMDCs were stimulated with antibiotics in the presence or absence of the caspase inhibitor Z-VAD-FMK. IL-1β secretion was measured in supernatants after 6 h of stimulation. Data are expressed as the mean ± SD from three independent experiments all performed in triplicates.
- FIGURE 3.
Polymyxin B, tyrothricin, gramicidin, and neomycin activate NLRP3 through different ways. A, LPS-primed wild type BMDCs were stimulated with antibiotics in serum-free buffer with or without 75 mM KCl and NaCl. IL-1β secretion was measured in supernatants after 6 h (B). Wild-type and P2X7-deficient BMDCs were primed with LPS and exposed to antibiotic. IL-1β secretion was measured in supernatants after 6 h of stimulation. ATP was used as control. C and D, LPS-primed wild type BMDCs were treated with cytochalasin D, N-acetyl cysteine (NAC), and CA-07-Me for 30 min. Pretreated cells were stimulated with antibiotics. IL-1β secretion was measured in supernatants after 6 h of stimulation. Data are expressed as the mean ± SD from three independent experiments all performed in triplicate.
Additional Files
Data Supplement
Files in this Data Supplement:
- Supplemental Figures 1-2 (PDF, 156 Kb) -
Description:
Figure 1. Antibiotics-induced IL-1 β production depends on LPS priming
Figure 2. Antibiotcs induced IL-1β secretion other immune cells and neutrophil influx
- Supplemental Figures 1-2 (PDF, 156 Kb) -
Description:
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