Sphingosine kinase 2 signaling modulates altered expression profile of host genome in a murine model of Pseudomonas aeruginosa pneumonia.

We present the set of pseudomonas responsive genes activated in the WT mice in comparison with Sphk2-/- thereby revealing the genes that could contribute to the protection seen in Sphk2-/- mice. Methods: Lung mRNA profiles of 8 week old male wild-type (WT) and Sphingosine kinase 2 knock out (Sphk2-/-) mice were generated by deep sequencing, in triplicate, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two way ANOVA (ANOVA). qRT–PCR validation was performed using SYBR Green assays Genetic deletion of Sphk2-/-resisted alteration of host pulmonary genome by PA infection of the lungs promoting its own virulence thereby conferring significant protection against PA pneumonia. Overall design: Age- and weight-matched male WT, Sphk1-/-, and Sphk2-/- mice were administrated a single intratracheal infusion of sterile PBS or PA103 in P (1x106 CFU/mouse). After 24 h of treatment, lung tissue was collected. Lung tissue was perfused with PBS prior to harvesting from the mice, and processed immediately after harvesting. Whole lung tissue was used to isolate total RNA using microRNeasy® kit (Qiagen, Maryland, Cat no. 74004).