OMIP-016
Characterization of antigen-responsive macaque and human T-cells
Cytometry PART A, Volume 83A, Issue 2, 182-184 (2013)
Sabrina Guenounou, Nathalie Bosquet, Claudia J. Dembek, Roger Le Grand, Antonio Cosma
Description: The present panel was optimized to assess the quality and phenotype of antigen-specific CD4 and CD8 T cells in both cynomolgus macaques and humans. The use of an identical protocol for specimens collected in the two species allows for an immediate translation of research from the macaque model to humans. Our protocol works well with cryopreserved and freshly collected PBMC. Following the fixing and permeabilization procedure, we introduced a freezing step that breaks the experimental procedure and allows the shipment of freshly stimulated and permeabilized samples to facilities equipped with instruments able to measure ten distinct fluorophores. Our procedure is thus adapted to multicenter studies where stimulation is performed on fresh PBMC and flow cytometry acquisition is done in a centralized facility.
Species: Human, Cynomolgus macaque
Tissue: Peripheral blood
Cell Subset: T cell, T memory, Th1
Keyword: Activation, Cryopreserved pbmc
Machine: BD LSR2
Panel details
| Marker | Species | Clone | Fluorophore | Purpose | Co-expressions groups | Antigen Density |
|---|---|---|---|---|---|---|
| CD40L | Cynomolgus macaque | TRAP1 | FITC | Cytokines/Chemokine/Activation marker | T cell, activation | Med |
| CD45RA | Cynomolgus macaque | L48 | PE-Cy7 | Differentiation | T cell, T memory | Med |
| CD3 | Cynomolgus macaque | SP34-2 | APC-Cy7 | T cell subset | T cell, T memory, Th1 | High |
| CD4 | Cynomolgus macaque | L200 | PerCP-Cy5.5 | T cell subset | T cell, T memory, Th1 | High |
| MIP-1 beta | Cynomolgus macaque | D21-1351 | PE | Cytokines/Chemokine/Activation marker | T cell, activation | Med |
| IFN-gamma | Cynomolgus macaque | B27 | V450 | Cytokines/Chemokine/Activation marker | T cell, Th1 | High |
| TNF | Cynomolgus macaque | Mab11 | Alexa Fluor 700 | Cytokines/Chemokine/Activation marker | T cell, Th1 | High |
| CD8a | Cynomolgus macaque | RPA-T8 | V500 | T cell subset | T cell | High |
| Dead cells | Cynomolgus macaque | Live/Dead Fix Blue | viability | |||
| IL-2 | Cynomolgus macaque | MQ1-17H12 | APC | Cytokines/Chemokine/Activation marker | T cell, Th1 | Med |
