ArticlePloS one2026
Comparison between enzyme-linked immunospot assay and intracellular cytokine flow cytometry assay of cytomegalovirus-specific T-cell response in healthy participants.
Article in PloS one, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Human cytomegalovirus (HCMV) usually establishes a lifelong latent infection after primary infection. Reactivation occurs sporadically and is controlled by cell-mediated immune response (CMIR). Monitoring of CMIR against CMV is mandatory in immunocompromised patients to adjust immunosuppressive drugs to prevent serious end-organ damage after CMV reactivation. Intracellular staining (ICS) and enzyme-linked immunospot (ELISPOT) quantifying CMV-specific T-cells are generally used as surrogate markers for CMIR against CMV. Whether the results of these 2 methods correlate well is not known. This study compared the numbers of CMV-specific T-cells identified by ELISPOT and ICS in healthy adult volunteers. Correlation with CMV serological status was explored. Thirty peripheral blood samples from healthy individuals were quantified for IFN-γ-producing cells after stimulation with whole CMV and IE1 using ICS, and IFN-γ-secreting cells after stimulation with whole CMV lysate and IE1 peptide pool using ICS and IFN-γ-secreting cells by ELISPOT. Anti-CMV IgG levels were analyzed concomitantly using a chemiluminescent microparticle immunoassay. There were 30 healthy participants, 15 (50%) male, with a mean age of 37.8 (± 7.6) years. Twenty-eight (93.3%) were seropositive against CMV. The CMV-specific CD3+ cells, as measured by ICS, were highly correlated with the spot numbers obtained by the ELISPOT, irrespective of CMV antigens used (whole CMV, r = 0.7677, p < 0.0001; IE1, r = 0.6516, p < 0.0001). The numbers of CMV-specific CD3+ cells quantified by IE1 stimulation by these 2 assays were statistically correlated with anti-CMV IgG levels (ICS, r = 0.5070, p = 0.004; ELISPOT, r = 0.4384, p = 0.015). Among the 30 participants, CMV-specific T cells were detected in all participants (100%), including the two seronegative individuals. The present study demonstrated that CMV-specific T-cells measured by ICS and ELISPOT assays were well correlated, suggesting that these assays could be used to monitor CMV-specific T-cells. CMV IgG levels may reflect prior CMV infection and CMV-specific cell-mediated immunity (CMIR) in immunocompetent individuals.
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