Transcriptome Analysis of Human Peripheral Blood Mononuclear Cells Exposed to Lassa Virus and to the Attenuated Mopeia/Lassa Reassortant 29 (ML29), a Vaccine Candidate

Zapata, Juan Carlos; Carrion, Ricardo; Patterson, Jean L.; Crasta, Oswald; Zhang, Yan; Mani, Sachin; Jett, Marti; Poonia, Bhawna; Djavani, Mahmoud; White, David M.; Lukashevich, Igor S.; Salvato, Maria S.

Due to the high number of interferon-related genes involved in the LASV and ML29 gene profiles, they were selected for validation by RT-PCR. 24 hour post-exposure (hpe) RNA samples, from 3 donors, and 3 time points used in the microarray test, were evaluated by the Interferon and Receptor PCR Array Kit (Human Interferon and Receptor array, Cat# No. PAHS-064, SABiosciences, Frederick, MD). This platform tested 84 interferon-related genes and 5 housekeeping genes for data normalization. Briefly, 1 mg of RNA from PBMC was reversetranscribed in a 20-mL reaction volume into first-strand cDNA using SuperArray's ReactionReady First Strand cDNA Synthesis Kit (Cat. No C-01, SABiosciences) containing random and oligo dT primers. After mixing the cDNA with RT2 SYBR Green/ ROX qPCR Master Mix (Cat. No PA-012, SABiosciences), 10 mL of the cDNA mixture was dispensed into a 96 well format plate RT 2 profiler PCR array (Cat. No PAHS-064, SAbioscience).