Identification of a novel autoantigen in inflammatory bowel disease by protein microarray

Vermeulen, Nathalie; Beéck, Katrijn Op De; Vermeire, Séverine; Steen, Kristel Van; Michiels, G; Ballet, Vera; Rutgeerts, Paul; Bossuyt, Xavier

doi:10.1002/ibd.21508

Cited by 33 publications

(20 citation statements)

References 30 publications

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“…However, we detected higher levels of anti-FAM84A antibodies in patients with acute inflammatory CNS diseases compared with MS or noninflammatory diseases, concomitant to increased overall IgG levels. This finding is consistent with the presence of anti-FAM84A antibodies in patients with inflammatory bowel disease (32) and indicates that FAM84A is an intracellular antigen that might elicit strong secondary immune reactivity when released as cellular debris.…”

Section: Resultssupporting

confidence: 86%

Distinct oligoclonal band antibodies in multiple sclerosis recognize ubiquitous self-proteins

Brändle

Obermeier

Şenel

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

157

110

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Oligoclonal Ig bands (OCBs) of the cerebrospinal fluid are a hallmark of multiple sclerosis (MS), a disabling inflammatory disease of the central nervous system (CNS). OCBs are locally produced by clonally expanded antigen-experienced B cells and therefore are believed to hold an important clue to the pathogenesis. However, their target antigens have remained unknown, mainly because it was thus far not possible to isolate distinct OCBs against a background of polyclonal antibodies. To overcome this obstacle, we copurified disulfide-linked Ig heavy and light chains from distinct OCBs for concurrent analysis by mass spectrometry and aligned patientspecific peptides to corresponding transcriptome databases. This method revealed the full-length sequences of matching chains from distinct OCBs, allowing for antigen searches using recombinant OCB antibodies. As validation, we demonstrate that an OCB antibody from a patient with an infectious CNS disorder, neuroborreliosis, recognized a Borrelia protein. Next, we produced six recombinant antibodies from four MS patients and identified three different autoantigens. All of them are conformational epitopes of ubiquitous intracellular proteins not specific to brain tissue. Our findings indicate that the B-cell response in MS is heterogeneous and partly directed against intracellular autoantigens released during tissue destruction. In addition to helping elucidate the role of B cells in MS, our approach allows the identification of target antigens of OCB antibodies in other neuroinflammatory diseases and the production of therapeutic antibodies in infectious CNS diseases. multiple sclerosis | oligoclonal bands | proteomics | transcriptomics | autoantigens M ultiple sclerosis (MS) is a severe inflammatory disease of the central nervous system (CNS) with a presumed autoimmune pathogenesis (1-4). Oligoclonal bands (OCBs) (5-7) of the cerebrospinal fluid (CSF) are the only established immunological biomarker of MS that has a diagnostic and prognostic relevance. OCBs are expanded Ig species that contain abundant somatic hypermutations, supposedly in response to sustained antigen stimulation (8-14). They are visualized by analytical immunoblotting on isoelectric focusing (IEF) gels, but so far, no particular target antigens could be assigned to distinct OCBs (15), because OCB quantities in diagnostic lumbar punctures are too low for direct antigen searches using biochemical techniques. Furthermore, OCBs are superimposed on a background of polyclonal antibodies, precluding differentiation between signals from OCB and background antibodies. Antibody cloning from single CSF B cells (16,17) is an elegant approach for obtaining matching heavy (H) to light (L) chains of antibodies, but it is impossible to know whether a particular antibody relates to an OCB or to the polyclonal background. Candidate target antigens detected with unfractionated CSF or with recombinant antibodies cloned from single CSF-resident B cells included lipids, proteins, combinatorial peptides, and viral products ...

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Section: Resultssupporting

confidence: 86%

Distinct oligoclonal band antibodies in multiple sclerosis recognize ubiquitous self-proteins

Brändle

Obermeier

Şenel

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

157

110

View full text Add to dashboard Cite

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“…Signal intensities were acquired using ProtoArray Prospector, a software tool developed for ProtoArray data by Invitrogen (14). We converted fluorescent intensities for the spot proteins (or autoantigens) to log 2 intensities and then normalized across the total 59 data sets by using the quantile normalization method (15).…”

Section: Analysis Of Protoarray Datamentioning

confidence: 99%

Identification of Novel Autoantibodies in Type 1 Diabetic Patients Using a High-Density Protein Microarray

et al. 2014

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Autoantibodies can facilitate diagnostic and therapeutic means for type 1 diabetes (T1DM). We profiled autoantibodies from serum samples of 16 T1DM patients, 16 type 2 diabetic (T2DM) patients, and 27 healthy control subjects with normal glucose tolerance (NGT) by using protein microarrays containing 9,480 proteins. Two novel autoantibodies, anti-EEF1A1 and anti-UBE2L3, were selected from microarrays followed by immunofluorescence staining of pancreas. We then tested the validity of the candidates by ELISA in two independent test cohorts: 1) 95 adults with T1DM, 49 with T2DM, 11 with latent autoimmune diabetes in adults (LADA), 20 with Graves disease, and 66 with NGT and 2) 33 children with T1DM and 34 healthy children. Concentrations of these autoantibodies were significantly higher in T1DM patients than in NGT and T2DM subjects (P < 0.01), which was also confirmed in the test cohort of children (P < 0.05). Prevalence of anti-EEF1A1 and anti-UBE2L3 antibodies was 29.5% and 35.8% in T1DM, respectively. Of note, 40.9% of T1DM patients who lack anti-GAD antibodies (GADA) had anti-EEF1A1 and/or anti-UBE2L3 antibodies. These were also detected in patients with fulminant T1DM but not LADA. Our approach identified autoantibodies that can provide a new dimension of information indicative of T1DM independent of GADA and new insights into diagnosis and classification of T1DM.Type 1 diabetes (T1DM) results from immune-mediated pancreatic b-cell destruction. Several autoantibodies, such as GAD65 antibody (GADA) (1), islet cell antibody (ICA) (2), protein tyrosine phosphatase antibody (IA-2 antibodies [IA-2A]) (3), and zinc transporter antibody (ZnT8A) (4), were identified and used for diagnosis and prediction of T1DM.However, the nature and mechanism of b-cell destruction in humans seem to be variable, which results in a broad spectrum of T1DM spanning from fulminant T1DM (5) to latent autoimmune diabetes in adults (LADA) (6). Fulminant T1DM is a rapidly progressing form of T1DM without evidence of autoimmunity, whereas LADA is autoimmune diabetes not requiring insulin at diagnosis. In most cases of LADA, which account for 10% of incident cases of diabetes in adults, b-cell function is severely impaired within 6 years, leading to insulin dependency (6). Fulminant T1DM is an important subtype of T1DM in Asian adults (7), accounting for 15%-20% of T1DM with ketosis or ketoacidosis in Japan (5). Moreover, T1DM in adults often lacks evidence of autoimmunity. The prevalence of autoantibodies associated with T1DM declines as the onset age increases, especially in the case of IA-2A (8) and ZnT8A

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“…Since its first description in 1984, several groups have tried intensively to identify the autoantigenic targets of PAB [24,[34][35][36][37][38]. Hence, the simultaneous report of the discovery of glycoprotein 2 (GP2) as a CrD-specific pancreatic autoantigen by two groups -that of Stöcker's and our group in 2008 was a remarkable coincidence [39][40][41][42].…”

Section: Identification Of Crd-specific Pancreatic Autoantigensmentioning

confidence: 99%

Crohn’s disease specific pancreatic antibodies: clinical and pathophysiological challenges

Roggenbuck¹,

Reinhold

Schierack

et al. 2014

Clinical Chemistry and Laboratory Medicine

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Crohn's disease (CrD) and ulcerative colitis (UC) are the main inflammatory bowel diseases (IBD). IBDspecific humoral markers of autoimmunity in the form of autoantibodies have been reported first in the late 1950s by demonstrating the occurrence of autoimmunity in UC, while humoral autoimmunity in CrD can be traced back to the 1970s. Ever since, the pathophysiological role of autoimmune responses in IBDs has remained poorly understood. Notwithstanding, autoreactive responses play a major role in inflammation leading to overt IBD. In CrD, approximately 40% of patients and < 20% of patients with UC demonstrate loss of tolerance to antigens of the exocrine pancreas. Glycoprotein 2 (GP2) has been identified as a major autoantigenic target of the so-called pancreatic antibodies. The previously unsolved contradiction of pancreatic autoreactivity and intestinal inflammation in IBD was elucidated by demonstrating the expression of GP2 at the site thereof. Intriguingly, GP2 has been reported to be a receptor on microfold cells of intestinal Peyer's patches, which are believed to represent the origin of CrD inflammation. The development of immunoassays for the detection of antibodies to GP2 has paved the way to investigate the association of such antibodies with the clinical phenotype in CrD. Given the recently discovered immunomodulating role of GP2 in innate and adaptive intestinal immunity, this association can shed further light on the pathophysiology of IBD. In this context, the association of anti-GP2 autoantibodies as novel CrD-specific markers with the clinical phenotype in CrD will be discussed in this review.

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Identification of a novel autoantigen in inflammatory bowel disease by protein microarray

Abstract: We identified FAM84A as a novel autoantigen in IBD. (Inflamm Bowel Dis 2011;).

Cited by 33 publications

References 30 publications

Distinct oligoclonal band antibodies in multiple sclerosis recognize ubiquitous self-proteins

Distinct oligoclonal band antibodies in multiple sclerosis recognize ubiquitous self-proteins

Identification of Novel Autoantibodies in Type 1 Diabetic Patients Using a High-Density Protein Microarray

Crohn’s disease specific pancreatic antibodies: clinical and pathophysiological challenges

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