Cloning of HCV-specific T cells after the second stimulation with or without CD137 enrichment*
| . | Not enriched . | CD137 enriched . |
|---|---|---|
| Total no. of wells | 480 | 480 |
| Percentage of specific T cells prior to cloning | 0.11 | 0.11 |
| Initial cell no./well | 4 | 4 |
| Total no. of wells with positive cell growth (%) | ∼ 75 (16) | ∼ 250 (52) |
| No. of wells tested by pMHC-multimer | 55 | 64 |
| % of multimer+ clones/CD8+ clone | 15 | 67 |
| % of multimer+ cells in parallel bulk culture | 12 | 62 |
| Overall specific cloning efficiency, %* | 2.3 | 35 |
| . | Not enriched . | CD137 enriched . |
|---|---|---|
| Total no. of wells | 480 | 480 |
| Percentage of specific T cells prior to cloning | 0.11 | 0.11 |
| Initial cell no./well | 4 | 4 |
| Total no. of wells with positive cell growth (%) | ∼ 75 (16) | ∼ 250 (52) |
| No. of wells tested by pMHC-multimer | 55 | 64 |
| % of multimer+ clones/CD8+ clone | 15 | 67 |
| % of multimer+ cells in parallel bulk culture | 12 | 62 |
| Overall specific cloning efficiency, %* | 2.3 | 35 |
Cloning was performed 24 hours after the second stimulation in medium containing irradiated allogenous B-LCLs and PBMCs as feeder cells and IL-2 and IL-15. Plates were evaluated by pMHC-multimer staining 20 days after cloning.
Specific cloning efficiency was determined as the percentage of wells with specific cell growth in relation to the total number of wells.