Analysis PipelineOur pipeline of longitudinal mass cytometry data analysis is based on three successive clusterings. Cells sharing similar phenotypes were gathered into cell clusters using SPADE. Then cell clusters displaying similar phenotypes were grouped into phenotypic families.
The steps of the longitudinal mass cytometry data analysis are displayed. As the first analysis step, single cells from FCS files were grouped into clusters sharing similar phenotype using the SPADE algorithm. NK cells were identified on the resulting SPADE tree as CD3- CD8+ cells. As the second analysis step, NK cell clusters sharing the same categories of marker expression were regrouped into phenotypic families. Finally, after these two successive clusterings, and as the last analysis step, discriminant analyses were used to determine which phenotypic families best distinguished between post-prime and post-boost immune response, and to define the phenotypic signature of each response.