Evaluation of cytomegalovirus‐specific T‐cell reconstitution in patients after various allogeneic haematopoietic stem cell transplantation using interferon‐γ‐enzyme‐linked immunospot and human leucocyte antigen tetramer assays with an immunodominant T‐cell epitope

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Abstract
Cytomegalovirus (CMV) infection is a major complication for patients who received allogeneic haematopoietic stem cell transplantation (HSCT). Accurate monitoring of CMV‐specific T‐cell reconstitution is required for appropriate decision on treatment, such as anti‐viral drugs, which have adverse effects. Although human leucocyte antigen (HLA) tetramer and interferon‐γ‐enzyme‐linked immunospot (IFN‐γ‐ELISPOT) assays have been used to measure CMV‐specific T cells, detailed comparison of these assays and kinetics of anti‐CMV T‐cell reconstitution between reduced‐intensity transplantation (RIST) and conventional HSCT has not yet been performed. In this study, we performed prospective comparative monitoring of CMV‐specific T cells using HLA tetramer and IFN‐γ‐ELISPOT assays with a single immunodominant CMV495 peptide in 28 HLA‐A*0201 and 9 HLA‐A*0206 patients after various allogeneic HSCTs. The IFN‐γ‐ELISPOT assay was more sensitive for evaluation of functional T cells than the HLA tetramer assay, and CMV‐specific T cells were reconstituted earlier in patients who received RIST without anti‐thymocyte globulin (ATG) than those receiving RIST with ATG or conventional HSCT. The threshold level for protection from CMV reactivation was estimated as over 1 × 106 cells/l peripheral blood with the IFN‐γ‐ELISPOT assay. These results demonstrate that the IFN‐γ‐ELISPOT assay with CMV495 provides more accurate evaluation on CMV immunity in HLA‐A*0201 and ‐A*0206 patients, and may be useful for determining timing of various treatments.

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