Vδ2neg γδ T-cell lines are able to lyse CMV-infected cells in vitro. (A) Monolayers of uninfected MRC5 fibroblasts or infected with TB40E or VHLE clinical CMV strains were cocultured at indicated E:T ratios for 4 hours at 37°C with Vδ2neg γδ T polyclonal cell lines raised from CMV-seropositive healthy donors (n = 8, ), from a CMV-seropositive transplantation recipient (■) at 2 early time points after transplantation and CMV-seronegative healthy donors (n = 9, ). Lysis of uninfected cells as control is shown (□). Results are the mean of specific lysis of culture duplicates. CV was always less than 15%. (B) Vδ2neg γδ T-cell lines were cocultured with CMV-infected fibroblasts in the absence or presence of blocking anti-TCR-γδ and anti-CD3 antibodies. Uninfected cell lysis (white bar), control as lysis with no blocking (gray bar), TCR blocking (dark gray bar), and CD3 blocking (black bar). (C) IFN-γ secretion detected in the culture supernatants after 6-hour stimulation coculture of Vδ2neg γδ T-cell line with CMV-infected fibroblasts at 10:1 E:T in the absence () or presence of anti-CD3 (□), anti-NKG2D, anti–TCR-γδ blocking (), and TCR/CD3/NKG2D (■).