Nuceloprotein pSLA-I tetramer staining of tissues from influenza vaccinated Babraham Pigs.

<p>Babraham pig 625 (left panel of 21 plots) and 650 (right panel of 21 plots) received H5N1 S-FLU intranasally and inactivated H1N1 virus [A/Swine/Spain/SF11131/2007] with montanide adjuvant intramuscularly, followed by a boost at day 25 using the same preparation. Pigs were culled at day 38 (day 13 post boost) and blood, bronchoalveolar lavage (BAL) and tracheobronchial lymph nodes (TBLNs) harvested and frozen as single cell suspensions. Tetramer staining was performed on thawed cells from the blood, BAL and TBLN using a no tetramer control, and staining with Irrelevant and nucleoprotein peptide tetramers. The sequences for the nucleoprotein peptides are shown. Irrelevant tetramers: SLA-1*14:02-AFAAAAAAL, SLA-2*11:04-AGAAAAAAI (pig 625) and SLA-2*11:04-GAGGGGGGI (pig 650). Gating strategy: lymphocytes, single cells, viability (Vivid^neg) CD3⁺ CD14^neg then CD8β⁺ CD4⁺ and displayed as CD8β versus tetramer (<b><a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1007017#ppat.1007017.s007" target="_blank">S1 Fig</a></b>).