Author: 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.
Title: Transcriptome Analysis of Human Peripheral Blood Mononuclear Cells Exposed to Lassa Virus and to the Attenuated Mopeia/Lassa Reassortant 29 (ML29), a Vaccine Candidate Document date: 2013_9_12
ID: 0epeljaf_27
Snippet: Whole blood from healthy donors was collected in heparin tubes, mixed with 20 ng/ml lipopolysaccharide (LPS Cat No-tlrlekpls, InvivoGen, San Diego, CA) as down-regulation control for thrombomodulin [45] , MEM as non-stimulated control and ML29 virus. Then this mix was brought to 3 ml with RPMI 10% FBS and incubated for 4, 8, or 24 hours. For each time-point cells were stained with phycoerytrhin-conjugated mouse anti-human thrombomodulin monoclona.....
Document: Whole blood from healthy donors was collected in heparin tubes, mixed with 20 ng/ml lipopolysaccharide (LPS Cat No-tlrlekpls, InvivoGen, San Diego, CA) as down-regulation control for thrombomodulin [45] , MEM as non-stimulated control and ML29 virus. Then this mix was brought to 3 ml with RPMI 10% FBS and incubated for 4, 8, or 24 hours. For each time-point cells were stained with phycoerytrhin-conjugated mouse anti-human thrombomodulin monoclonal antibody (PE-CD141, BD Biosciences, San Jose, CA) and fluorescein isothiocyanate mouse anti-human CD14+ antibody (FITC-CD14+ BD Biosciences, San Jose, CA). Cells were collected (10,000 events per sample) using a FACSCalibur cytometer (BD Biosciences) and the data were analyzed using FlowJo 8.8.4 software (Tree Star, San Carlos, CA). The monocyte population was gated and percentages of double positive cells (CD14+ and CD141+) were determined.
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