OMIP‐087: Thirty‐two parameter mass cytometry panel to assess human CD4 and CD8 T cell activation, memory subsets, and helper subsets

Abstract

We developed a highly reproducible 32‐marker mass cytometry panel able to measure all canonical immune lineages and perform detailed characterization of both CD4 and CD8 T cells in human peripheral blood mononuclear cells. In this panel, we identify six different T cell memory subsets, as well as markers of activation, cell cycling, and survival. In addition, this panel classifies all major CD4 T cell helper subsets. This panel enables detailed monitoring of CD4 and CD8 T cells in the context of infectious disease, cancer or autoimmunity with limited patient sample use. Detailed methods for standardization and optimization of the panel can be found in Supporting Information.

Dominant CD4+ T cell receptors remain stable throughout antiretroviral therapy-mediated immune restoration in people with HIV

“…Optimization and standardization of the mass cytometry panel was detailed previously. 126 PBMC were thawed and rested overnight in R10+ at 37°C with average viability of 89% (71–97%). 10 7 cells/mL PBMC suspension was prepared in Ca 2+ Mg 2+ free PBS (Rockland, Cat# MB008) and cell-ID Cisplatin 195 (Fluidigm Cat# 201195) was added.…”

“…Plots were ArcSinh transformed with a co-factor of 5 and then gates were set using either unstained or antibody backbone-stained controls. 126 The unstained tube was used to confirm there was no environmental metal contamination and to appropriately identify CD45 + cells. The antibody backbone tube which consisted of all major lineage markers (CD45, CD3, CD4, CD8, TCRγδ, CD19, CD14, CD56 and CD16) was used to set gates downstream of these lineages markers.…”