ZIKV-specific CD8+ T Cell Analysis

Approximately 1–5 × 10⁶ brain/spleen/blood cells were transferred to U-bottom 96-well microtiter plates and were incubated for 20 min (4°C in the dark) with PE-conjugated H-2D^b tetramers for ZIKV E_294–302 (34 (link)) and subsequently stained for an additional 20 min (4°C in the dark) for relevant cell-surface markers. Next, the cells were centrifuged, washed, fixed in 1% PFA and finally resuspended in PBS and stored at 4°C until flow cytometric analysis. Cell samples were analyzed using FACS LSRFortessa cytometer (BD Biosciences), and the data was analyzed using FlowJo software version 10 (Tree Star).