5. Present Investigations
5.4 Paper 4
DNA-fragmentation is a source of bactericidal activity against Pseudomonas aeruginosa.
Background
Cystic fibrosis (CF) is a genetic disorder that affects many organs but the lung disease is the major cause of morbidity and mortality in these patients. The characteristics of CF lung disease include chronic bacterial infections (especially Pseudomonas aeruginosa), inflammatory exacerbations, and highly viscous sputum of the lower airways. Long-lasting and dysregulated inflammatory responses in CF airways leads to accumulation of immune cells like neutrophils, that eventually succumb, resulting in abundant extracellular DNA (eDNA). This eDNA is a major contributor to the viscous mucus seen in this disease and is the basis for use of recombinant human DNaseI as a treatment for reducing the viscoelasticity of the sputum for efficient clearance of the airway congestion.
Aim
- To investigate the effects of DNase I treatment in a murine model of acute P.
aeruginosa airway infection.
- To determine the molecular properties of eDNA and possible roles for DNA-fragmentation, in executing bactericidal activity against P. aeruginosa.
Results and conclusions
DNase I treatment of P. aeruginosa infected mice enhanced their survival and decreased the bacterial load in the BALF and lung tissue. The treatment significantly decreased the pro-inflammatory cytokines IL-6 and TNF-α in the lung tissue.
The eDNA isolated form the BALF of the DNase I treated mice showed increased fragmentation with accumulation of smaller fragments of sizes around and below 200 bp.
By using synthetic DNA fragments of decreasing lengths, we showed that there is a size threshold for the DNA fragments to exhibit bactericidal activity and this activity was possibly because of the chelation of divalent cations from the surface of Gram-negative bacterial outer membrane.
The findings suggest a novel host defense strategy that could potentially be employed to treat P. aeruginosa infections, circumventing mechanisms involved in resistance against conventional antibiotics.
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