Caspofungin enhances antifungal immunity in human blood through transcriptional reprogramming
Caspofungin enhances antifungal immunity in human blood through transcriptional reprogramming
Human peripheral blood was collected from healthy volunteers with written informed consent. This study was conducted in accordance with the Declaration of Helsinki and all protocols were approved by the Ethics Committee of the University Hospital Jena (permit number: 3639-12/12). After 8 hours of infection, human RNA was stabilized in PAXgene Blood RNA tubes (PreAnalytiX) and extracted using the PAXgene Blood RNA Isolation Kit (PreAnalytiX) according to the manufacturer's instructions. RNA concentration was measured with a NanoDrop OneC spectrophotometer (Thermo Fisher Scientific), and RNA integrity was assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies).   Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.