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.

Monocytes were isolated from purified PBMC by plastic adherence and immature dendritic cells (DC) were generated from them after 5 days culture in supplemented RPMI. Briefly, the method for generating DC was to plate human PBMC at 1.67610 7 cells/2 ml/well in a 6-well plate in RPMI, 10% fetal calf serum. After 2 hrs, non-adherent cells were removed and the adherent monocytes were fed with 2.5 ml RPMI containing 20 ng/ml GM-CSF (Peprotech, Rocky Hill NJ), and 20 ng/ml IL-4 (Peprotech). After 5 days of incubation, cells were harvested, analyzed by flow cytometry, and processed for incubation with virus. On average, 22% of the cells were differentiated, monocytederived DC (DR+, CD11c+). [38] .