{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T10:15:49Z","timestamp":1780049749587,"version":"3.53.1"},"posted":{"date-parts":[[2026,5,26]]},"group-title":"Immunology","reference-count":40,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,26]],"date-time":"2026-05-26T00:00:00Z","timestamp":1779753600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000065","name":"NINDS","doi-asserted-by":"crossref","award":["NS064571"],"award-info":[{"award-number":["NS064571"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"crossref"}]},{"name":"NCI","award":["U01CA271410"],"award-info":[{"award-number":["U01CA271410"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,26]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Interferon-\u03b3 (IFN\u03b3) signaling is prominent in inflammatory CNS microenvironments across many neurological disorders, but the neuronal peptides presented on HLA class I under these conditions and their functional consequences for CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cells remain incompletely defined. Here we combine human iPSC-derived human neural aggregates (HNAs), HLA class I immunoprecipitation coupled to LC-MS\/MS immunopeptidomics, and microfluidic co-culture assays to map IFN\u03b3-induced neuronal antigen presentation and test antigen-specific cytotoxicity. IFN\u03b3 stimulation induced robust HLA class I expression in HNAs and enabled recovery of a canonical 8-12-mer class I ligandome enriched for 9-mers. Neuron-restricted expression of a synapsin-driven polyepitope cassette yielded presentation of defined neoantigen 9-mer peptides on donor HLA class I molecules and, in the presence of IFN\u03b3, elicited activation of autologous antigen-specific CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cells and consequent antigen-dependent neurite injury. Across four donors, comparative immunopeptidomics identified large IFN\u03b3-unique neural peptide repertoires distinct from matched fibroblasts and revealed a consistent enrichment of predicted high-affinity binders on HLA-B allotypes. Finally, \u03b22-microglobulin deletion ablated peptide recovery, and neuron-restricted \u03b22-microglobulin reconstitution enabled identification of neuron-derived peptides, including peptides derived from neurofilament light (NEFL) that were shared across donors and presented on multiple HLA allotypes. Together, these data provide an integrated platform for neuronal autoantigen discovery and functional validation and support a model in which IFN\u03b3-driven neuronal HLA class I presentation creates an HLA-B-weighted epitope landscape that can be recognized by autoreactive cytotoxic CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cells.\n                <\/jats:p>","DOI":"10.64898\/2026.05.22.727243","type":"posted-content","created":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T00:40:24Z","timestamp":1779842424000},"source":"Crossref","is-referenced-by-count":0,"title":["Human neurons stimulated with IFN\u03b3 present HLA class I-restricted autoantigens to cytotoxic CD8\n                  <sup>+<\/sup>\n                  T cells"],"prefix":"10.64898","author":[{"given":"Benjamin DS","family":"Clarkson","sequence":"first","affiliation":[{"name":"Translational Neuroimmunology Laboratory, Mayo Clinic, Rochester, Minnesota, USA"},{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, USA"},{"name":"Center for Multiple Sclerosis and Autoimmune Neurology, Mayo Clinic, Rochester, Minnesota, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanna","family":"Pucci","sequence":"additional","affiliation":[{"name":"Translational Neuroimmunology Laboratory, Mayo Clinic, Rochester, Minnesota, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramila Barun","family":"Shrestha","sequence":"additional","affiliation":[{"name":"Translational Neuroimmunology Laboratory, Mayo Clinic, Rochester, Minnesota, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kiran K","family":"Mangalaparthi","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Remya","family":"Raja","sequence":"additional","affiliation":[{"name":"Department of Immunology, Mayo Clinic, Phoenix, Arizona, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marion","family":"Curtis","sequence":"additional","affiliation":[{"name":"Department of Immunology, Mayo Clinic, Phoenix, Arizona, USA"},{"name":"Mayo Clinic Comprehensive Cancer Center, Mayo Clinic, Phoenix, Arizona, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akhilesh","family":"Pandey","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota, USA"},{"name":"Manipal Academy of Higher Education, Manipal, Karnataka, India"},{"name":"Center for Individualized Medicine, Mayo Clinic, Rochester, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3889-8739","authenticated-orcid":false,"given":"Charles L","family":"Howe","sequence":"additional","affiliation":[{"name":"Translational Neuroimmunology Laboratory, Mayo Clinic, Rochester, Minnesota, USA"},{"name":"Center for Multiple Sclerosis and Autoimmune Neurology, Mayo Clinic, Rochester, Minnesota, USA"},{"name":"Division of Experimental Neurology, Mayo Clinic, Rochester, Minnesota, USA"},{"name":"Department of Neurology, Mayo Clinic, Rochester, Minnesota, USA"},{"name":"Department of Immunology, Mayo Clinic, Rochester, Minnesota, 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