Page 53 - The autoimmune hypothesis of narcolepsy and its unexplored clinical features M.S. Schinkelshoek

Figure 3.3. H1N1-HA275-287-specific T cell lines are predominantly CD4+. These fluorescence-activated cell sorting (FACS) plots show the gating strategy of two T cell lines in different patients (A-C and D-F). Living single cells are selected in the FSC-SSC plot (A and D). In the next plot CD3+ cells are selected (B and E), which show a high percentage of CD4+ cells in the last plot (C and F).
To assess HLA-DQ6 restriction, T cell proliferation experiments were performed with irradiated allogeneic PBMCs of different haplotypes: HLA-DQA1*01:02/ DQB1*06:03, HLA-DQA1*03:01/DQB1*03:02 (HLA-DQ8) or a mix of HLA- DQA1*01:02/DQB1*06:02 and HLA-DQA1*01:02/DQB1*06:03 next to those expressing HLA-DQA1*01:02/DQB1*06:02. T cell proliferation was only observed in the presence of HLA-DQ6-expressing PBMCs and the H1N1- HA275-287 peptide. Furthermore, when T cell proliferation experiments were performed with irradiated allogeneic PBMCs expressing HLA-DQ6, an anti- HLA-DQ mAb was able to block T cell proliferation of the generated TCCs. An anti-HLA-class I mAb blocked T cell proliferation considerably less effectively, whereas anti-HLA-DR and –DP mAbs did not affect T cell proliferation (Figure 3.4A-B). These experiments confirm that the detected H1N1-HA275-287-specific CD4+ T cell proliferation is HLA-DQ6-restricted.
H1N1 reactivity in CD4+ T cells
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