Analysis of the EBV-specific CTL response and EBV DNA load in the peripheral blood of IM patients.
(A-C) Functional analysis of EBV-specific CTL responses in acute IM patients 2 (A) and 1 (B), and 3 healthy donors (C) using ELISPOT assay. PBMCs from these individuals were stimulated with peptide epitopes from early lytic, late lytic, and latent antigens, and the interferon γ response was measured in ELISPOT assays as described previously.4 The following CTL epitopes were used in this study: restricted through HLA A2—GLCTLVAML (GLC), SLVIVTTFV (SLV), ILIYNGWYA (ILI), VLQWASLAV (VLQ), VLTLLLLLV (VLT), LIPETVPYI (LIP), QLTPHTKAV (QLT), LLDFVRFMGV (LLD), YLQQNWWTL (YLQ), YLLEMLWRL (YLL), LLVDLLWLL (LLV), TLLVDLLWL (TLL), LTAGFLIFL (LTA), CLGGLLTMV (CLG); HLA B7—RPPIFIRRL (RPP); and HLA A24—RYSIFFDY (RYS), TYGPVFMCL (TYG). For acute IM patients, ELISPOT analysis was conducted during the acute phase of infection and at different time intervals after diagnosis (AD). The results are expressed as spot-forming cells (SFCs) per 106 PBMCs. NT indicates not tested. (D) EBV DNA load in the peripheral blood of IM patients 2 (left) and 1 (right) at diagnosis and after diagnosis. EBV DNA load was measured as described elsewhere.5 (E) EBV epitope–specific ex vivo cytotoxic T-cell activity in peripheral blood lymphocytes from IM donors using peptide-sensitized (1 μg/mL) phytohemagglutinin blasts as targets. Peptide epitopes used in these assays are shown on the x-axis. Results are expressed as percent specific lysis. E/T ratio indicates effector-target ratio.
