Identification of influenza-specific T-cells in the bronchoalveolar lavage from Babraham pigs infected with pandemic H1N1 swine influenza virus.

<p>Babraham pigs were either left uninfected (741) or infected intranasally with pandemic H1N1 [A/sw/Eng/1353/09] (742, 744, 745). The pigs were culled on day 0 (741), 5 (744) or 14 (742 and 745) post infection. (<b>A</b>) 200,000 bronchoalveolar lavage cells from pig 745 (infected, day 14 cull) were incubated alone, with 10^-5M peptide, or virus for 16–18 h. A Babraham kidney cell line was included in each well (15,000 per well) to act as antigen presenting cells. All conditions were performed in duplicate and spot forming cells (SFCs) detected by IFNγ ELISPOT and displayed as mean +SEM and scaled (X5) to 10⁶ BAL cells. (<b>B</b>) Irrelevant and nucleoprotein peptide-SLA-1*14:02 and SLA-2*11:04 tetramer staining was performed on thawed bronchoalveolar lavage samples and the percentage of tetramer⁺ cells of CD8β cells displayed in red. Nucleoprotein peptide sequences are shown. Irrelevant tetramers were SLA-1*14:02-AFAAAAAAL and SLA-2*11:04-AGAAAAAAI. 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>).