Simultaneous quantification of human cytomegalovirus (HCMV)-specific CD4+ and CD8+ T cells by a novel method using monocyte-derived HCMV-infected immature dendritic cells

Laura Lozza, Daniele Lilleri, Elena Percivalle, Chiara Fornara, Giuditta Comolli, Maria Grazia Revello, Giuseppe Gerna

Research output: Contribution to journalArticlepeer-review

Abstract

Immature dendritic cells (DC) infected with an endotheliotropic (Huv+) and leukotropic (Leuk+) human cytomegalovirus (HCMV) strain were used as a stimulus to determine functional HCMV-specific CD4+ and CD8+ T cells. Infected DC were co-cultured with autologous peripheral blood mononuclear cells and both arms of T cell activation were determined by intracellular flow cytometry analysis of IFN-γ production. Efficient stimulation of HCMV-specific CD4+ and CD8+ T cell responses was achieved using DC productively infected with Huv+ Leuk+ VR1814 strain. On the contrary, a negligible CD8+ T cell response was obtained when HCMV strains unable to infect DC, or DC pulsed with inactivated viral antigen, were used. HCMV specificity of the T cell response was confirmed in 46 HCMV-seropositive and 8 HCMV-seronegative healthy subjects. A cut-off was established to discriminate between immune and nonimmune subjects. The novel ex vivo assay enables the simultaneous evaluation of HCMV-specific CD4+ and CD8+ T cell responses and maybe a useful tool for monitoring HCMV-specific T cell activity in immunocompromised transplanted patients.

Original languageEnglish
Pages (from-to)1795-1804
Number of pages10
JournalEuropean Journal of Immunology
Volume35
Issue number6
DOIs
Publication statusPublished - Jun 2005

Keywords

  • Cytokine flow cytometry
  • Dendritic cells
  • HCMV
  • Specific T cells

ASJC Scopus subject areas

  • Immunology

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