Analysis of HLA-A24-restricted CMVpp65 peptide-specific CTL with HLA-A*2402-CMVpp65 tetramer.

Developing precise and efficient methods of monitoring immune responses against cytomegalovirus (CMV) infection in immunocompromised transplantation patients is important. With the aim of optimizing the monitoring strategy, an HLA-A24-CMVpp65 tetramer-based analysis of CMVpp65 peptide-specific CTL lines was performed. Previously, the HLA-A24-restricted CTL epitope of CMVpp65 matrix protein was identified (QYDPVAALF aa 341-349). In the present study, CMVpp65 (aa 341-349) peptide-specific CTL lines were obtained from PBLs of 12 HLA-A24+ healthy donors by two stimulations with peptide-pulsed dendritic cells (DC). Among 12 CTL lines, 9 showed HLA-A*2402-CMVpp65 tetramer staining, which was found to be strongly co-related to the amount of IFN-gamma produced by CMVpp65 peptide-restimulated CTL lines (r=0.943, P<0.001). These results suggested that HLA-A*2402-CMVpp65 tetramer staining was an efficient way to monitor immune responses against CMV infection in HLA-A24+ immunocompromised hosts.