ZIKV-Specific T Cell Response Quantification

Cellular response was assessed in spleens through ELISPOT using the IFN-γ ELISPOT set (BD Biosciences, USA) as previously described (26 (link)) 0.5x10⁶ cells/well were stimulated with 2 µg/mL of recombinant ZIKV E protein (Native Antigen, UK). The anti-CD3 and anti-CD28 antibodies (1 µg/mL each) (BD Bioscience, USA) were used as positive control. Unstimulated cells were used as baseline. Spots were quantified using AID iSpot FluoroSpot Reader System and AID EliSpot Version 7.0 software (AID-Autoimmun Diagnostika GMBH, Germany). Assays were performed in duplicate, baseline values were subtracted from all samples, and the results were expressed as the average number of spots forming cells (SFCs) per 10⁶ cells.