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.

Vero E6 cells (ATCC, CRL-1586) were cultured in Dulbecco's modified Eagle's medium (DMEM, GIBCO-BRL) supplemented with 10% fetal bovine serum (FBS, GIBCO-BRL), 1% penicillinstreptomycin, and L-glutamine (2 mM); incubated at 37uC in 5% CO 2 for 24 hours. Vero cell monolayers were infected with LASV and ML29 at a multiplicity of infection (MOI) of 0.01 and incubated 1 hour at 37uC in 5% CO 2 , washed with PBS and then covered in DMEM 2% FBS. Supernatants were collected at 48 and 72 hours post infection, titrated in Vero cells and stored at 270uC at 10 7 PFU/ml. Human PBMC were obtained by apheresis from healthy volunteers followed by Ficoll-hypaque isolation [37] and resuspended in RPMI 1640 medium with 10% heat-inactivated human AB serum (Sigma).