PMC:3359311 / 10426-12381 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/3359311","sourcedb":"PMC","sourceid":"3359311","source_url":"http://www.ncbi.nlm.nih.gov/pmc/3359311","text":"PBMCs from CF patients do not produce increased amounts of IL-1β upon inflammasome stimulation\nPrevious studies have shown that the loss of CFTR results in increased NF-κB activity and pro-inflammatory cytokine secretion [4], [5], [32], [36], [37]. To further examine this relationship, PBMCs from CF patients and healthy adult controls were stimulated with PAO1, LPS+ATP, and LPS+Poly(dA:dT), to activate the NLRC4, NLRP3, and AIM2 inflammasomes, respectively. By 24 hours of stimulation, CF PBMCs did not produce increased amounts of IL-1β (Fig. 5a) or IL-8 (Fig. 5b) when compared to healthy controls. However, we did notice a transient decrease (P\u003c0.001) in the amount of IL-1β produced by CF cells in response to LPS+ATP at 6 hours (data not shown). Stimulation of PBMCs with P. aeruginosa that lacks exsA (PAO1ΔexsA), a key regulator of type III secretion, produced three-fold less IL-1β compared to the parental PAO1 strain by 24 hours (Fig. 5a). Inflammasome stimulation without priming did not result in any IL-1β production in either CF or control PBMCs. Contrary to our hypothesis, these results indicate that PBMCs from CF patients do not display increased production of IL-1β or IL-8 with inflammasome activation nor do they suggest any increased basal or induced NF-κB activity. These results are consistent with our observation that caspase-1 activity is not different between CF and control PBMCs (Fig. 4).\n10.1371/journal.pone.0037689.g005 Figure 5 PBMCs from CF patients do not produce increased amounts of IL-1β.\nPBMCs from CF patients (n = 17–20) and healthy controls (n = 15–19) were primed with LPS (10 ng/ml) overnight and stimulated with P. aeruginosa PAO1 (MOI = 1), P. aeruginosa PAO1 lacking exsA (MOI = 1), ATP (5 mM), or Poly(dA:dT) (1 µg/ml) for 24 hours. P. aeruginosa lacking exsA was used as a type III secretion control in comparison with wild-type P. aeruginosa. Supernatants were assayed for (A) IL-1β and (B) IL-8.\n\nN","divisions":[{"label":"Title","span":{"begin":0,"end":94}},{"label":"Figure 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