Expression of PPARγ and paraoxonase 2 correlated with Pseudomonas aeruginosa infection in cystic fibrosis

Phoebe E Griffin, Louise F Roddam, Yvonne C Belessis, Roxanne Strachan, Sean Beggs, Adam Jaffe, Margaret A Cooley, Phoebe E Griffin, Louise F Roddam, Yvonne C Belessis, Roxanne Strachan, Sean Beggs, Adam Jaffe, Margaret A Cooley

Abstract

The Pseudomonas aeruginosa quorum sensing signal molecule N-3-oxododecanoyl-l-homoserine lactone (3OC(12)HSL) can inhibit function of the mammalian anti-inflammatory transcription factor peroxisome proliferator activated receptor (PPAR)γ, and can be degraded by human paraoxonase (PON)2. Because 3OC(12)HSL is detected in lungs of cystic fibrosis (CF) patients infected with P. aeruginosa, we investigated the relationship between P. aeruginosa infection and gene expression of PPARγ and PON2 in bronchoalveolar lavage fluid (BALF) of children with CF. Total RNA was extracted from cell pellets of BALF from 43 children aged 6 months-5 years and analyzed by reverse transcription-quantitative real time PCR for gene expression of PPARγ, PON2, and P. aeruginosa lasI, the 3OC(12)HSL synthase. Patients with culture-confirmed P. aeruginosa infection had significantly lower gene expression of PPARγ and PON2 than patients without P. aeruginosa infection. All samples that were culture-positive for P. aeruginosa were also positive for lasI expression. There was no significant difference in PPARγ or PON2 expression between patients without culture-detectable infection and those with non-Pseudomonal bacterial infection, so reduced expression was specifically associated with P. aeruginosa infection. Expression of both PPARγ and PON2 was inversely correlated with neutrophil counts in BALF, but showed no correlation with other variables evaluated. Thus, lower PPARγ and PON2 gene expression in the BALF of children with CF is associated specifically with P. aeruginosa infection and neutrophilia. We cannot differentiate whether this is a cause or the effect of P. aeruginosa infection, but propose that the level of expression of these genes may be a marker for susceptibility to early acquisition of P. aeruginosa in children with CF.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Relative gene expression of PPARγ…
Figure 1. Relative gene expression of PPARγ in BALF from CF patients with and without culture-defined P. aeruginosa infection.
Horizontal bars represent median values. Significance assessed by Mann–Whitney U test. P. aeruginosa pos  =  P. aeruginosa infection detected by culture (>105 cfu/mL BALF). P. aeruginosa neg  =  P. aeruginosa undetectable by culture.
Figure 2. Relative gene expression of PPARγ…
Figure 2. Relative gene expression of PPARγ in BALF from CF patients with P. aeruginosa detected by RT–qPCR.
Horizontal bars represent median values. Significance assessed by Kruskal–Wallis analysis with Dunn’s post test. P. aeruginosa neg  =  no detectable 16S rRNA product in PCR. P. aeruginosa low  =  product detected Cp>30; P. aeruginosa med  =  product detected Cp 20–30; P. aeruginosa high  =  product detected Cp<20.
Figure 3. Low PPARγ gene expression is…
Figure 3. Low PPARγ gene expression is associated with P. aeruginosa infection but not with presence of other pathogens.
Horizontal bars represent median values. Significance compared with no pathogen group assessed by Kruskal–Wallis analysis with Dunn’s post test. No pathogen: no bacterial or fungal pathogens detected by culture. non-Pa bacteria: infection with any bacteria other than P. aeruginosa without detectable P. aeruginosa and A. fumigatus; A. fumigatus: infection with A. fumigatus without detectable P. aeruginosa and with or without other bacteria; P. aeruginosa: infection with P. aeruginosa with or without other pathogens. Infection defined as >105 cfu/mL BALF.
Figure 4. Low PON2 gene expression is…
Figure 4. Low PON2 gene expression is associated with P. aeruginosa infection but not with other pathogens.
Horizontal bars represent median values. Significance compared with no pathogen group assessed by Kruskal–Wallis analysis with Dunn’s post test. No pathogen: no bacterial or fungal pathogens detected by culture. non-Pa bacteria: infection with any bacteria other than P. aeruginosa with undetectable P. aeruginosa and A. fumigatus; A. fumigatus: Aspergillus fumigatus infection without detectable P. aeruginosa and with or without other bacteria; P. aeruginosa: P. aeruginosa infection with or without other pathogens. Infection defined as >105 cfu/mL BALF.

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