EBNA 3c [LLDFVRFMGV] peptide-specific IFN-γ production of HLA-A*0201⁺ EBV-reactive T cells.

An EBV-reactive T-cell line of an HLA-A*0201⁺donor was generated for 2 weeks, and the peptide-specific CD3⁺CD8⁺ T cells were determined by tetramer analysis or the intracellular IFN-γ assay. (A) Demonstrates that 0.4% of the CD3⁺CD8⁺ bound the HLA tetramer (EBNA3c [LLDFVRFMGV]). (B) Similarly, 0.3% of the CD3⁺CD8⁺ cells responded with IFN-γ production after secondary stimulation with EBNA3c (LLDFVRFMGV) peptide-pulsed PBMCs. (C, D) Only 0.02% of aliquots of the same T cells responded with IFN-γ production after secondary stimulation with autologous PBMCs, pulsed with EBNA 3a [QAKWRLQTL], which binds the HLA-B*0801 allele not expressed by this donor, or in response to another HLA-A*0201-binding peptide, the NLVPMVATV peptide from the CMV pp65 antigen, respectively. (E) Demonstrates that 3.9% of the CD3⁺CD8⁺ cells responded with intracellular IFN-γ production after secondary stimulation with autologous EBV BLCL.