Table 2.
Parameters selected for the ICS consensus assay.
| Assay step | Recommendations |
|---|---|
| Thawing Temperature | 37°C |
| Cell Count Method | Automated |
| Viability method Pre-culture | PI/Trypan Blue/Other |
| Viability Post-Culture | Live/Dead |
| Cell number/well (during stimulation) | 1,000,000 |
| Overnight Resting | None |
| Extracellular Panel | Live/Dead staining, CD4/CD8 |
| Intracellular Panel | CD3/CD40L/IFNγ/IL2/TNFα |
| Read-Out | Polypositive CD3+/CD4+ or CD3+/CD8+ polypositive cells (CD40L, IFNγ, IL2, TNFα) |
| Minimal number of acquired cells | Acquire at least 75,000 Live CD3+CD4+ parent cells Optimally 100,000 CD4+ T cells should be acquired |
ICS, intracellular cell staining; IFN, interferon; IL, interleukin; PI, propidium iodide; TNF, tumor necrosis factor.