Evaluation of Epstein-Barr Virus Latent Membrane Protein 2 Specific T-Cell Receptors Driven by T-Cell Specific Promoters Using Lentiviral Vector
Figure 3
Expression of TCR α and β chains in the HSB2 cells and PBMCs. (a) The PBMCs were incubated with lentivirus containing EBV-LMP2 TCR α and β chains driven by T cell-specific promoters at MOIs of 1 and 10. Three days aftertransduction, cells were collected and stained for CD3 mobilization as a measure of TCR α chain expression. (b) Photographs of flow cytometric analysis for TCR α chain expression in HSB2 cells (top) and PBMC cells (bottom). The cells were infected with recombinant lentivirus or control empty lentivirus at MOI of 10 for 3 days. TCR staining was performed by using anti-TCR α chain antibody (prepared from our lab) followed by PE-labeled second antibody. (c) Percentage of TCR α chain-positive cells in transduced HSB2 cells 1 week and 7 weeks after infection at MOI of 10. (d) Percentage of TCR α chain-positive cells in transduced CD3+ cells 1 week and 7 weeks after infection. PBMCs stimulated with IL-2 plus OKT3 for 24 hr were infected with the lentivirus at an MOI of 10. After 1 week, the PBMCs were analyzed by FACS for TCR α chain expression and then maintained in culture with IL-2 for 7 weeks for reanalysis for TCR α chain-positive cells.
