Fig. 1: Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1. | Nature Communications

Fig. 1: Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1.

From: Enhanced influenza A H1N1 T cell epitope recognition and cross-reactivity to protein-O-mannosyltransferase 1 in Pandemrix-associated narcolepsy type 1

Fig. 1: Overall experimental study design for T-cell epitope discovery in Pandemrix-associated NT1.

As a first step, influenza A (H1N1) virus HA, NA, and NP peptide T cell recognition was tested with spleen cells from Pandemrix-vaccinated HLA-DQ6.2 mice, restimulated with pools of 5 peptides each (15-mers). Pools that stimulated IFN-γ or IL-2 expression were broken up, and single peptides tested either with spleen cells from additional Pandemrix-vaccinated HLA-DQ6.2 mice, or PBMC from Pandemrix-vaccinated HLA-DQB1*0602 positive individuals. As a second step, recognition of single peptides was then tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls. As a third step, influenza A (H1N1) T cell peptides that showed increased stimulation of IFN-γ or IL-2 secretion in patients vs. controls were validated and mapped. Cross-reactive T cell self-epitopes were predicted by BLAST against human proteome, and recognition tested with PBMC from pediatric Pandemrix-associated NT1 patients, and healthy Pandemrix-vaccinated controls.

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