Figure 2. Replication of NiV in the individual subpopulations of PBMC.

Sorted cells were inoculated with NiV in three independent experiments. Total of 10⁵cells per subpopulation were inoculated with 10⁴ PFU of NiV in one 1 ml volume. The infectivity in cell supernatants was determined by plaque titration on Vero 76 cells. Fig. 2.A Comparison of virus yield from stimulated and non-stimulated PBMC subpopulations at 48 hrs post inoculation. Black filled columns represent mean of pfu/ml in supernatant collected from non-stimulated cells, the grey filled columns represent mean of pfu/ml in supernatant collected from stimulated lymphocytes or monocytes. Purity of the individual cell preparations was in the following range: B cells - 75 to 85%, CD6+ T cells - 90 to 95%, CD4+ T cells - 85%, CD3+CD8+ T cells - 95%, NK cells – 80 to 85%, monocytes - close to 100%. Fig. 2.B Presence of genomic RNA at indicated time points post inoculation in cell lysates from monocytes inoculated with live NiV (solid line) or with gamma-irradiated NiV (dashed line). Fig. 2.C NiV yield in supernatants of sorted T lymphocytes at 24 hrs post inoculation. Fig. 2.D Detection of infectivity in supernatant harvested from monocytes at 24 and 48 hrs post inoculation with NiV (gray columns), and detection of viral RNA in cells inoculated with live virus (solid black line) or with gamma-irradiated NiV (black dashed line).