TY - JOUR
T1 - Simultaneous quantification of human cytomegalovirus (HCMV)-specific CD4+ and CD8+ T cells by a novel method using monocyte-derived HCMV-infected immature dendritic cells
AU - Lozza, Laura
AU - Lilleri, Daniele
AU - Percivalle, Elena
AU - Fornara, Chiara
AU - Comolli, Giuditta
AU - Revello, Maria Grazia
AU - Gerna, Giuseppe
PY - 2005/6
Y1 - 2005/6
N2 - 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.
AB - 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.
KW - Cytokine flow cytometry
KW - Dendritic cells
KW - HCMV
KW - Specific T cells
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U2 - 10.1002/eji.200526023
DO - 10.1002/eji.200526023
M3 - Article
C2 - 15902686
AN - SCOPUS:20844437647
VL - 35
SP - 1795
EP - 1804
JO - European Journal of Immunology
JF - European Journal of Immunology
SN - 0014-2980
IS - 6
ER -