Snapshot assessment of RNA-expression in severely burned patients using the PAXgene Blood RNA system: a pilot study.

ABSTRACT

Severe burn induces destabilization of the immune system and the likelihood of multiple organ dysfunction syndrome. Current studies focus on RNA-expression analyses of immune system cells, however, the present methods of analysis are complex, potentially altered by artefacts and therefore not feasible for routine analyses. The new PAXgene Blood RNA System provides "snapshot" analysis of RNA by immediate cell lysis and prevention of RNA-degradation. Using this system the aim of this study was to analyse intracellular cytokine RNA-expression under clinical conditions. Whole blood samples (PAXgene tubes) of nine severely burned patients were drawn at admission and 6, 12, 24, 48 and 72h after trauma during routine treatment. Four healthy individuals served as control. Analysis of RNA-expression of TNF-alpha as pro-inflammatory and IL-10 as anti-inflammatory mediator was performed by RT-PCR. The RNA-expression of TNF-alpha was increased at 72h after burn. The increase occurred mainly in surviving patients. In contrast, RNA-expression of IL-10 was elevated already at 24h and the difference between surviving and deceased patients occurred earlier. We demonstrate for the first time a "snapshot" analysis of cytokine RNA-expression in severely burned patients under routine conditions. The results correspond well to current hypothesis of posttraumatic MODS development.