title: Simultaneous quantification of human cytomegalovirus (HCMV)-specific CD4+ and CD8+ T cells by a novel method using monocyte-derived HCMV-infected immature dendritic cells.
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-gamma 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 may be a useful tool for monitoring HCMV-specific T cell activity in immunocompromised transplanted patients.
authors.affiliation: Servizio di Virologia, IRCCS Policlinico San Matteo, Pavia, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Lozza
authors.forename: Daniele
authors.initials: D
authors.lastname: Lilleri
authors.forename: Elena
authors.initials: E
authors.lastname: Percivalle
authors.forename: Chiara
authors.initials: C
authors.lastname: Fornara
authors.forename: Giuditta
authors.initials: G
authors.lastname: Comolli
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Revello
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gerna
mesh_terms: D018791:CD4 Lymphocyte Count; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D018920:Coculture Techniques; D003587:Cytomegalovirus / Q000276:immunology*; D003713:Dendritic Cells / Q000502:physiology / Q000821:virology*; D005434:Flow Cytometry; D006801:Humans; D016867:Immunocompromised Host; D007371:Interferon-gamma / Q000096:biosynthesis; D008213:Lymphocyte Activation; D018655:Lymphocyte Count; D015203:Reproducibility of Results
chemical_list: D007371:Interferon-gamma