Functional analysis of cytomegalovirus-specific T lymphocytes compared to tetramer assay in patients undergoing hematopoietic stem cell transplantation

In order to evaluate whether we could predict reactivation of CMV by monitoring the number of CMV-specific cytotoxic T-lymphocytes (CTL), tetramer analysis was performed in 37 patients who underwent hematopoietic stem cell transplantation (HSCT). The results disclosed that the mean number of CMV-specific CTL at day 30 did not differ among patients who developed CMV antigenemia (22/μl) and those who did not (12/μl). Serial tetramer analysis showed that 21% of the patients had >10/μl CMV-specific CTL at the first detection of CMV antigenemia and 67% of the patients had more than 10/μl CMV-specific CTL at the onset of CMV disease. Intracellular staining upon stimulation by CMV lysates and peptide in patients with CMV colitis revealed that both IFN-γ producing CD4+ and CD8+ lymphocytes were suppressed at the onset of CMV colitis (1.6 and 8/μl), which increased with recovery of the disease (19 and 47/μl). These data suggest that it is difficult to predict CMV reactivation solely by the number of CMV-specific CTL. We suggest that additional functional analysis by intracellular cytokine assay may be useful for immunomonitoring against CMV.