Statistical significance (* 0.05, ** 0.01, *** 0.001) is shown. in these individuals were characterized in depth by ex lover vivo immunophenotyping and transcriptome analysis of relevant T-cell subsets. Further examination of these individuals included CSF markers of swelling and neurodegeneration and a detailed characterization with respect to demographic, medical, and MRI features. Results By screening CSF-infiltrating CD4+ T cells from 105 individuals with MS against seven long-known myelin and five recently explained GDP-l-fucose synthase peptides, we recognized GDP-l-fucose synthase and myelin oligodendrocyte glycoprotein (35-55) responder individuals. Immunophenotyping of ODM-203 CSF and combined blood samples in these individuals revealed a significant expansion of an effector memory space (CCR7? CD45RA?) CD27? Th1 CD4+ cell subset in GDP-l-fucose synthase responders. Subsequent transcriptome analysis of this subset demonstrated manifestation of Th1 and cytotoxicity-associated genes. Individuals with different intrathecal T-cell specificities also differ concerning swelling- and neurodegeneration-associated biomarkers, imaging findings, manifestation of HLA class II alleles, and seasonal distribution of the time of the lumbar puncture. Conversation Our observations reveal an association between autoantigen reactivity and features of disease heterogeneity that strongly supports an important part of T-cell specificity in MS pathogenesis. These data have the potential to improve patient classification in medical practice and to guide the development of antigen-specific tolerization strategies. Multiple Rabbit Polyclonal to TOR1AIP1 sclerosis (MS) is considered an immune-mediated demyelinating disease of the CNS.1 There is general agreement about the relevance of B cells,2 CD8+ T cells,3,4 and particularly CD4+ ODM-203 T cells in MS pathogenesis. However, despite the fact that myelin-specific CD4+ T cells can exacerbate MS5 and that specific T cells can induce disease in different animal models of MS,6,7 there is less consensus about the relevance of CD4+ T-cell specificity. Disease heterogeneity is definitely a hallmark of MS and likely stems from the interplay of a complex genetic background8,9 and environmental risk factors.10 Although it is well explained that disease heterogeneity encompasses composition of cells lesions, imaging findings, clinical presentation, disease course, and treatment responsiveness, the factors contributing to this heterogeneity, including a putative involvement of T-cell specificity, are not clear. From studies in Experimental Autoimmune Encephalomyelitis (EAE), the most commonly used animal model in MS,6,11 it is well known that both the rodent strain, we.e., the genetic background, and the antigen used to induce EAE influence disease course, severity, lesion distribution, and pathologic mechanisms,7,12,13 linking T-cell specificity and disease heterogeneity. The recognition of the prospective antigens in neurologic autoimmune diseases14-16 has led to great progress in understanding, diagnosing, and treating these diseases. To better understand and treat MS, researchers possess investigated CD4+ T-cell specificity for years, albeit with less success than expected. The difficulties in obtaining CNS-infiltrating CD4+ T cells, the overall low antigen avidity of selfCantigenCreactive T cells, and the limited information about T cellCspecific autoantigens in MS have hampered research progress. In addition to the long-known myelin peptides,17 few T cellCspecific autoantigens have been recognized so far. We recently recognized GDP-l-fucose synthase (GDP-L-FS)-derived peptides as putative autoantigens in human being leukocyte antigen (HLA)-DRB3* individuals with MS.18 To evaluate the relevance of CD4+ T-cell specificity in MS, we analyzed their link with features of disease heterogeneity. We recognized individuals with different CSF-infiltrating CD4+ T-cell reactions against myelin and GDP-L-FS peptides and characterized them by ex lover vivo immunophenotyping of CSF and combined blood samples as well as with respect to demographic, medical, and MRI features. Methods Patient Samples Paired CSF and blood samples were collected from 105 untreated individuals with MS, only CSF from 11 control individuals (CPs), and 10 individuals with myelin oligodendrocyte glycoprotein antibodyCassociated disease (MOGAD) bad for anti-AQP4 antibodies and only peripheral blood mononuclear cells (PBMCs) from four healthy donors (HDs) (eTable 1, links.lww.com/NXI/A590). Eighty-four individuals with MS (80%) experienced by no means been treated, whereas 21 (20%) were previously treated but regarded as untreated at the time of lumbar puncture (eTable 1, links.lww.com/NXI/A590). Individuals and settings were recruited from your NIMS-Neuroimmunology and MS Study Section, Department of Neurology, University Hospital Zurich. MS diagnosis was based on the revised McDonald criteria.19 Standard Protocol Approvals, Registrations, and Patient Consents The Cantonal Ethics Committee of Zurich approved the study procedures (EC-No. 2013-0001). Informed consent was obtained from all patients. CSF and Serum Steps Albumin quotients and CSF-specific oligoclonal bands were analyzed as previously reported.20 Intrathecal Ig synthesis (Ig[loc]) was calculated according to Reiber.21 Cell Culture and Stimulation PBMCs were freshly isolated using Ficoll (Eurobio, Germany) density-gradient centrifugation. CSF-infiltrating CD4+ T cells were expanded with phytohaemagglutinin in a single round ODM-203 of stimulation using a previously reported protocol.