Polyfunctional Analysis of Human Cytomegalovirus (HCMV)-Specific CD4+ and CD8+ Memory T-Cells in HCMV-Seropositive Healthy Subjects Following Different Stimuli

Elisa Gabanti, Francesca Bruno, Chiara Fornara, Stefano Bernuzzi, Daniele Lilleri, Giuseppe Gerna

Research output: Contribution to journalArticlepeer-review


Purpose: Following primary human cytomegalovirus (HCMV) infection, both humoral and T-cell-mediated immune responses develop in immunocompetent subjects. However, while antibodies may be measured by different methodologies, the T-cell-mediated response remains to be analyzed in its polyfunctional aspects, in view of defining (following different stimuli) the optimal assay to monitor the HCMV-specific T-cell response in HCMV-seropositive subjects.

Methods: In a group of 30 HCMV-seropositive adults, T-cell response revealed by the HCMV-infected dendritic cell (iDC) stimulus was compared with those given by the HCMV-infected cell lysate (iCL), and by a 34-peptide pool (PP).

Results: All HCMV-seropositive subjects showed presence of both HCMV-specific CD4+ and CD8+ T-cells in peripheral blood following iDC stimulation. One subject did not respond to PP. As compared to iDC, the number of HCMV-specific stimulated T-cells/μl blood was slightly lower for iCL (P = 0.195) and significantly lower for PP (P = 0.001). Polyfunctional analysis of the T-cell response indicated that the lower number of CD4+ T-cells stimulated by iCL was due to the bifunctional (IFN-γ+ TNF-α+) and CD40L-negative T-cell reduction, while the reduction in specific PP-stimulated CD8+ T-cells was attributable to the reduction in tri-(IFN-γ+ TNF-α+ IL2+), bi-(IFN-γ+ TNF-α+) and mono-(IFN-γ+) functional T-cells. In addition, 15/30 (50 %) subjects showed a CD4+ cross-response to PP, and 11/30 (37 %) a CD8+ cross-response to iCL.

Conclusions: HCMV-specific stimulus given by iDC is not significantly different from that of iCL on CD4+ and is significantly superior to that of PP on CD8+ T-cells. However, iCL may contribute significantly to CD8+, and PP to CD4+ T-cell stimulation.

Original languageEnglish
Pages (from-to)999-1008
Number of pages10
JournalJournal of Clinical Immunology
Issue number8
Publication statusPublished - Nov 5 2014


  • dendritic cells
  • Human cytomegalovirus
  • memory T-cells
  • peptide pool
  • polyfunctional analysis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Medicine(all)


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