title: High-Density Peptide Microarray Analysis of IgG Autoantibody Reactivities in Serum and Cerebrospinal Fluid of Multiple Sclerosis Patients.
abstract: Intrathecal immunoglobulin G (IgG) synthesis and oligoclonal IgG bands in cerebrospinal fluid (CSF) are hallmarks of multiple sclerosis (MS), but the antigen specificities remain enigmatic. Our study is the first investigating the autoantibody repertoire in paired serum and CSF samples from patients with relapsing-remitting MS (RRMS), primary progressive MS (PPMS), and other neurological diseases by the use of high-density peptide microarrays. Protein sequences of 45 presumed MS autoantigens (e.g.MOG, MBP, and MAG) were represented on the microarrays by overlapping 15mer peptides. IgG reactivities were screened against a total of 3991 peptides, including also selected viral epitopes. The measured antibody reactivities were highly individual but correlated for matched serum and CSF samples. We found 54 peptides to be recognized significantly more often by serum or CSF antibodies from MS patients compared with controls (pvalues <0.05). The results for RRMS and PPMS clearly overlapped. However, PPMS patients presented a broader peptide-antibody signature. The highest signals were detected for a peptide mapping to a region of the Epstein-Barr virus protein EBNA1 (amino acids 392-411), which is homologous to the N-terminal part of human crystallin alpha-B. Our data confirmed several known MS-associated antigens and epitopes, and they delivered additional potential linear epitopes, which await further validation. The peripheral and intrathecal humoral immune response in MS is polyspecific and includes antibodies that are also found in serum of patients with other diseases. Further studies are required to assess the pathogenic relevance of autoreactive and anti-EBNA1 antibodies as well as their combinatorial value as biomarkers for MS.
authors.affiliation: From the ‡University of Rostock, Department of Neurology, Division of Neuroimmunology, Gehlsheimer Str. 20, 18147 Rostock, Germany; §Steinbeis Transfer Center for Proteome Analysis, Schillingallee 70, 18057 Rostock, Germany; michael.hecker@rocketmail.com.
authors.forename: Michael
authors.initials: M
authors.lastname: Hecker
authors.affiliation: From the ‡University of Rostock, Department of Neurology, Division of Neuroimmunology, Gehlsheimer Str. 20, 18147 Rostock, Germany; §Steinbeis Transfer Center for Proteome Analysis, Schillingallee 70, 18057 Rostock, Germany;
authors.forename: Brit
authors.initials: B
authors.lastname: Fitzner
authors.affiliation: From the ‡University of Rostock, Department of Neurology, Division of Neuroimmunology, Gehlsheimer Str. 20, 18147 Rostock, Germany;
authors.forename: Matthias
authors.initials: M
authors.lastname: Wendt
authors.affiliation: ¶University of Rostock, Institute of Immunology, Schillingallee 70, 18057 Rostock, Germany;
authors.forename: Peter
authors.initials: P
authors.lastname: Lorenz
authors.affiliation: ¶University of Rostock, Institute of Immunology, Schillingallee 70, 18057 Rostock, Germany;
authors.forename: Kristin
authors.initials: K
authors.lastname: Flechtner
authors.affiliation: ¶University of Rostock, Institute of Immunology, Schillingallee 70, 18057 Rostock, Germany; ‖Gesellschaft für Individualisierte Medizin mbH (IndyMED), Lessingstr. 17, 18055 Rostock, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Steinbeck
authors.affiliation: From the ‡University of Rostock, Department of Neurology, Division of Neuroimmunology, Gehlsheimer Str. 20, 18147 Rostock, Germany;
authors.forename: Ina
authors.initials: I
authors.lastname: Schröder
authors.affiliation: §Steinbeis Transfer Center for Proteome Analysis, Schillingallee 70, 18057 Rostock, Germany; ¶University of Rostock, Institute of Immunology, Schillingallee 70, 18057 Rostock, Germany; ‖Gesellschaft für Individualisierte Medizin mbH (IndyMED), Lessingstr. 17, 18055 Rostock, Germany.
authors.forename: Hans-Jürgen
authors.initials: HJ
authors.lastname: Thiesen
authors.affiliation: From the ‡University of Rostock, Department of Neurology, Division of Neuroimmunology, Gehlsheimer Str. 20, 18147 Rostock, Germany;
authors.forename: Uwe Klaus
authors.initials: UK
authors.lastname: Zettl
mesh_terms: D000328:Adult; D000918:Antibody Specificity; D001323:Autoantibodies / Q000097:blood* / Q000134:cerebrospinal fluid* / Q000378:metabolism; D001324:Autoantigens / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000134:cerebrospinal fluid / Q000378:metabolism; D008297:Male; D046228:Microarray Analysis / Q000379:methods; D008875:Middle Aged; D020528:Multiple Sclerosis, Chronic Progressive / Q000276:immunology*; D020529:Multiple Sclerosis, Relapsing-Remitting / Q000276:immunology*; D010455:Peptides / Q000378:metabolism*
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; D007074:Immunoglobulin G; D010455:Peptides
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