Page 52 - The autoimmune hypothesis of narcolepsy and its unexplored clinical features M.S. Schinkelshoek

Chapter 3
287-specific T cell proliferative response was observed (Figure 3.2). To assess the specificity of the T cell lines against the Hcrt peptides, they were also stimulated with the Hcrt56-68 and Hcrt87-99 peptides. No reactivity to Hcrt56-68 or Hcrt87-99 was observed in T cell lines of either NT1 patients or controls (Figure 3.2), indicating the absence of cross-reactivity of H1N1-HA275-287-specific T cells with Hcrt56-68 and Hcrt87-99 in the context of HLA-DQ6. FACS analysis was performed on 13 HLA-DQ6-H1N1-HA275-287-specific T cell lines. Analyses revealed a dominance of CD4+ T cells in these T cell lines (Figure 3.3).
Figure 3.2. Overview of T cell responses to H1N1-HA275-287, Hcrt56-68 and Hcrt87-99 for narcolepsy type 1 patients (red) and healthy controls (green). The P-value results from a Mann-Whitney U-test.
T cell reactivity to H1N1-HA275-287 is HLA-DQ6-restricted
T cell clones (TCCs) were generated from the H1N1-HA275-287-reactive T cell lines of 12 NT1 patients by either limiting dilution (in T cell lines of 3 NT1 patients) or isolation using streptavidin-PE-HLA-DQ6- H1N1-HA275-287 tetramers (in T cell lines of 9 NT1 patients) and tested for reactivity against H1N1-HA275-287. In TCCs generated from all H1N1-HA275-287-specific CD4+ T cell lines, H1N1-HA275-287-specific clonal T cell responses were found (Table 3.3).
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