OX40 assay
Varicella-Zoster virus, VZV
EVENTS
Heparin blood was collected from participants pre-treatment and 7 days post-treatment and diluted 1:1 with Iscoves modified medium (Sigma-Aldrich). Each sample was tested for CD4 + T cell-specific responses towards Phytohaemagluttinin (PHA) (Sigma-Aldrich) and Varicella-Zoster virus (VZV) (Source-Bioscience). 2 µg/ml of antigen or 15 µg/ml of PHA was added to a 500 µl volume with appropriate negative control tubes. After incubation at 37 °C, 5% COSamples were gated to acquire 30,000 CD3 positive events, which were then negatively gated for CD14 and 7AAD. Quantification of CD3 + CD4 + CD25 + CD134 + events were calculated by setting gate coordinates on the media-only sample to equal 0.1% CD3 + CD4 + CD25 + CD134 + events as a percentage of all CD3 + CD4 + events. These coordinates were applied to subsequent antigen tubes to produce a percentage value of CD25 + CD134 + double-positive events