Autoimmunity to hypocretin and molecular mimicry to flu in type 1 narcolepsy

Type 1 narcolepsy (T1N) is caused by hypocretin/orexin (HCRT) neuronal loss. Association with the HLA DQB1*06:02/DQA1*01:02 (98% vs. 25%) heterodimer (DQ0602), T cell receptors (TCR) and other immune loci suggest autoimmunity but autoantigens are unknown. Onset is seasonal and associated with influenza A, notably pandemic 2009 H1N1 (pH1N1) infection and vaccination (Pandemrix). Peptides derived from HCRT and influenza A, including pH1N1, were screened for DQ0602 binding and presence of cognate DQ0602 tetramer-peptide-specific CD4⁺ T cells tested in 35 T1N cases and 22 DQ0602 controls. Higher reactivity to influenza pHA_273-287 (pH1N1 specific), PR8 (H1N1 pre-2009 and H2N2)-specific NP_17-31 and C-amidated but not native version of HCRT_54-66 and HCRT_86-97 (HCRT_NH2) were observed in T1N. Single-cell TCR sequencing revealed sharing of CDR3β TRBV4-2-CASSQETQGRNYGYTF in HCRT_NH2 and pHA_273-287-tetramers, suggesting molecular mimicry. This public CDR3β uses TRBV4-2, a segment modulated by T1N-associated SNP rs1008599, suggesting causality. TCR-α/β CDR3 motifs of HCRT_54-66-NH2 and HCRT_86-97-NH2 tetramers were extensively shared: notably public CDR3α, TRAV2-CAVETDSWGKLQF-TRAJ24, that uses TRAJ24, a chain modulated by T1N-associated SNPs rs1154155 and rs1483979. TCR-α/β CDR3 sequences found in pHA_273-287, NP_17-31, and HCRT_NH2 tetramer-positive CD4⁺ cells were also retrieved in single INF-γ-secreting CD4⁺ sorted cells stimulated with Pandemrix, independently confirming these results. Our results provide evidence for autoimmunity and molecular mimicry with flu antigens modulated by genetic components in the pathophysiology of T1N.