Computational protocol: Efficacy of a Plasmodium vivax Malaria Vaccine Using ChAd63 and Modified Vaccinia Ankara Expressing Thrombospondin-Related Anonymous Protein as Assessed with Transgenic Plasmodium berghei Parasites

Similar protocols

Protocol publication

[…] PBMCs were stimulated for 5 h in the presence of TRAP peptide pools as described above. Hepatic cellular responses were assessed from perfused livers that were digested with collagenase and treated with ammonium-chloride-potassium (ACK) buffer to lyse red blood cells. Phenotypic and functional analyses of CD8+ and CD4+ T cells were performed using the following antibody clones: anti-CD8 peridinin chlorophyll protein (PerCP)-Cy5.5 (clone 53-6.7), the nanocrystal eFluor 650-coupled anti-CD4 (GK1.5), anti-IFN-γ allophycocyanin (APC) (XMG1.2), anti-tumor necrosis factor alpha (TNF-α) eFluor 450 (MP6-XT22), and anti-interleukin 2 (IL-2) phycoerythrin (PE)-Cy 7 (JES6-5H4). Flow cytometric analyses were performed using an LSRII instrument (BD Biosciences, Oxford, United Kingdom). The frequencies of cells producing cytokines in the graphs represent data from which background from nonstimulated cells was subtracted. Data were analyzed with either FACSDiva or FlowJo software. Analysis of multifunctional CD8+ T cell responses was performed using Boolean analysis in FlowJo software, Pestle, and SPICE 4.0 (). […]

Pipeline specifications

Software tools FlowJo, SPICE
Application Flow cytometry
Organisms Vaccinia virus, Mus musculus, Plasmodium vivax, Plasmodium berghei, Pan troglodytes, Homo sapiens, Plasmodium falciparum
Diseases Malaria, HIV Infections