Fig. 5

RANTES is up-regulated in TBEV-infected human brain-derived cells. T98G (a, b), CCF-STTG1 (c, d), and SK-N-SH (e, f) cells and HPDAs (g and h) were inoculated with medium alone, TBEV, or UV-inactivated TBEV at an MOI of 1. Total RNA was extracted from cell lysates at 6, 24, 48, and 72 h post inoculation. RANTES mRNA was quantified by real-time PCR (a, c, e, and g), and results were normalized to GAPDH and expressed as fold induction over medium alone at 6 h post inoculation. Supernatants were harvested at 6, 24, 48, and 72 h post inoculation, and levels of RANTES (pg/mL) released were determined by ELISA (b, d, f, and h). Bars represent the means ± the standard deviations of three independent experiments. *P < 0.05 versus mock control. i TBEV replication kinetics in human brain cells. T98G, CCF-STTG1, and SK-N-SH cells and HPDAs were infected with TBEV at an MOI of 1. Supernatants were collected at indicated time points, and virus titers were determined by TCID₅₀ assay. The results are presented as the means ± standard deviations obtained from three independent experiments. j Neutralization of RANTES in T98G cell supernatants with specific anti-hRANTES Ab (αRANTES) reduced THP-1 cell migration. The same amount of normal goat IgG was used as controls. Data are expressed as the percentages of migrated cells in total cells added and presented as means ± the standard deviations of three independent experiments. *P < 0.05 versus control; ^# P < 0.05 versus TBEV-infected alone. Statistical analysis was performed by Student’s t test