An antisense peptide nucleic acid against Pseudomonas aeruginosa inhibiting bacterial-induced inflammatory responses in the cystic fibrosis IB3-1 cellular model system.
Discovery of novel antimicrobial agents against Pseudomonas aeruginosa able to inhibit bacterial growth as well as the resulting inflammatory response is a key goal in cystic fibrosis research. We report in this paper that a peptide nucleic acid (PNA3969) targeting the translation initiation region of the essential acpP gene of P. aeruginosa, and previously shown to inhibit bacterial growth, concomitantly also strongly inhibits induced up-regulation of the pro-inflammatory markers IL-8, IL-6, G-CSF, IFN-γ, IP-10, MCP-1 and TNF-α in IB3-1 cystic fibrosis cells infected by P. aeruginosa PAO1. Remarkably, no effect on PAO1 induction of VEGF, GM-CSF and IL-17 was observed. Analogous experiments using a two base mis-match control PNA did not show such inhibition. Furthermore, no significant effects of the PNAs were seen on cell growth, apoptosis or secretome profile in uninfected IB3-1 cells (with the exception of a PNA-mediated up-regulation of PDGF, IL-17 and GM-CSF). Thus, we conclude that in cell culture an antimicrobial PNA against P. aeruginosa can inhibit the expression of pro-inflammatory cytokines otherwise induced by the infection. In particular, the effects of PNA-3969 on IL-8 gene expression are significant considering the key role of this protein in the cystic fibrosis inflammatory process exacerbated by P. aeruginosa infection.
Department of Life Sciences and Biotechnology, Ferrara, Italy.
Giulia
G
Montagner
Department of Pathology and Diagnostics, University Hospital of Verona, Italy.
Valentino
V
Bezzerri
Department of Pathology and Diagnostics, University Hospital of Verona, Italy.
Giulio
G
Cabrini
Department of Life Sciences and Biotechnology, Ferrara, Italy.
Enrica
E
Fabbri
Department of Life Sciences and Biotechnology, Ferrara, Italy.
Monica
M
Borgatti
Department of Life Sciences and Biotechnology, Ferrara, Italy.
Ilaria
I
Lampronti
Department of Life Sciences and Biotechnology, Ferrara, Italy.
Alessia
A
Finotti
Department of Cellular and Molecular Medicine & Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, The Panum Institute, University of Copenhagen, Denmark. Electronic address: ptrn@sund.ku.dk.
Peter E
PE
Nielsen
Department of Life Sciences and Biotechnology, Ferrara, Italy. Electronic address: gam@unife.it.
Roberto
R
Gambari
D017209:Apoptosis / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation / Q000235:genetics; D003550:Cystic Fibrosis / Q000473:pathology*; D006801:Humans; D007249:Inflammation / Q000150:complications / Q000235:genetics / Q000382:microbiology; D016209:Interleukin-8 / Q000235:genetics; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D020135:Peptide Nucleic Acids / Q000235:genetics*; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*; D015854:Up-Regulation / Q000235:genetics
D016209:Interleukin-8; D016376:Oligonucleotides, Antisense; D020135:Peptide Nucleic Acids
Cystic fibrosis; IL-8; Peptide nucleic acids; Pseudomonas aeruginosa