Quantitative RT-PCR analyses of GAPdH, VZV ORF63 and ORF68 expression in VZV-infected monocytes, NK cells, NKT cells, B cells, CD4+ T cells, CD8+ T cells and HFLs.
Human PBMCs were infected with VZV as described in Fig 1, then uninfected- and VZV-infected monocytes, NK cells, NKT cells, B cells, CD4+ T cells and CD8+ T cells were sorted using flow cytometry. RNA was harvested and then analyzed by quantitative RT-PCR for ORF63 (grey bars), ORF68 (red bars) and GAPdH (green bars) transcripts. In addition, VZV-infected HFLs that were >90% VZV gE+ based upon flow cytometry analyses were harvested as a positive control for viral transcript expression. Results representative of sorted PBMCs from 3 different healthy individuals and from 3 independent HFL infections, with bar graphs representing average 1/Ct ± SD. Dotted line represents threshold for Ct values (1/36). All immune cell populations were >93% pure based upon flow cytometry sorting.