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

• Published in: European journal of immunology Vol. 35; no. 6; pp. 1795 - 1804
• Main Authors: Lozza, Laura, Lilleri, Daniele, Percivalle, Elena, Fornara, Chiara, Comolli, Giuditta, Revello, Maria Grazia, Gerna, Giuseppe
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY‐VCH Verlag 01-06-2005
• Subjects: CD4 Lymphocyte Count; CD8-Positive T-Lymphocytes - immunology; Coculture Techniques; Cytokine flow cytometry; Cytomegalovirus - immunology; Dendritic cells; Dendritic Cells - physiology; Dendritic Cells - virology; Flow Cytometry; HCMV; Humans; Immunocompromised Host; Interferon-gamma - biosynthesis; Lymphocyte Activation; Lymphocyte Count; Reproducibility of Results; Specific T cells
• ISSN: 0014-2980; 1521-4141
• DOI: 10.1002/eji.200526023

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 may be a useful tool for monitoring HCMV‐specific T cell activity in immunocompromised transplanted patients.