Secreted human Ro52 autoantibody proteomes express a restricted set of public clonotypes

Arentz, Georgia; Thurgood, Lauren A.; Lindop, Rhianna; Chataway, Tim; Gordon, Thomas P.

doi:10.1016/j.jaut.2012.07.003

Cited by 34 publications

(53 citation statements)

References 22 publications

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“…For example, mutated Sm-specific V-region clonotypic peptides might be used in targeted mass spectrometric platforms such as multiple reaction monitoring to quantitate specific V-region peptides in complex serum mixture, or to 10 monitor expression of anti-Sm clonal turnover in patients undergoing treatment [14,25]. A priority will be to sequence linked RNP/Sm autoantibody populations to test whether intermolecular sharing of autoreactive clonotypes holds true for the Sm/RNP autoantigen cluster.…”

Section: Discussionmentioning

confidence: 99%

“…MS data was analysed using Peaks Studio v7.0 software (Bioinformatics Solution Inc, Canada) using the following parameters: 2 ppm; product ion error tolerance of 0.01 Da; precursor charge state of +2 to +4; database search performed against a combined ImMunoGeneTics (IMGT) (http://www.imgt.org) and the Uniprot 2010-06 database as described previously [14].…”

Section: Protein Sequence Data Analysismentioning

confidence: 99%

“…Advances in mass spectrometric-based protein sequencing now allow heavy (H)-and (L)-chain V-region peptide maps to be generated for the first time from microgram quantities of autoantibodies derived from complex patient sera [12]. Recently, we have combined 2-dimensional gel electrophoresis (2-DE) of anti-Ro52/Ro60/La autoantibodies with de novo and database-driven V-region protein sequencing to show that humoral responses against these protein-RNA complexes are mediated by public (shared) autoreactive B cell clonotypes [13][14][15]. An early study reported oligoclonality and restricted V-region gene usage by anti-Sm hybridomas derived from MRL/lpr mice, raising the possibility of restricted anti-Sm clonality in human SLE [16].…”

Section: Introductionmentioning

confidence: 99%

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Serum SmD autoantibody proteomes are clonally restricted and share variable-region peptides

Kindi

Chataway

Gilada

et al. 2015

Journal of Autoimmunity

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Recent advances in mass spectrometry-based proteomic methods have allowed variable (V)-region peptide signatures to be derived from human autoantibodies present in complex serum mixtures. Here, we analysed the clonality and V-region composition of immunoglobulin (Ig) proteomes specific for the immunodominant SmD protein subunit of the lupus-specific Sm autoantigen. Precipitating SmD-specific IgGs were eluted from native SmD-coated ELISA plates preincubated with sera from six patients with systemic lupus erythematosus (SLE) positive for anti-Sm/RNP. Heavy (H)-and light (L)-chain clonality and V-region sequences were analysed by 2-dimensional gel electrophoresis and combined de novo database mass spectrometric sequencing. SmD autoantibody proteomes from all six patients with SLE expressed IgG1 kappa restricted clonotypes specified by IGHV3-7 and IGHV1-69 H-chains and IGKV3-20 and IGKV2-28 L-chains, with shared and individual V-region amino acid replacement mutations. Clonotypic sharing and restricted V-region diversity of systemic autoimmunity can now be extended from the Ro/La cluster to Sm autoantigen and implies a common pathway of anti-Sm autoantibody production in unrelated patients with SLE.3

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Section: Discussionmentioning

confidence: 99%

Section: Protein Sequence Data Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Serum SmD autoantibody proteomes are clonally restricted and share variable-region peptides

Kindi

Chataway

Gilada

et al. 2015

Journal of Autoimmunity

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“…We have recently developed a proteomic method that allows near full-length protein sequencing of epitope-specific populations of autoantibodies purified from whole serum [17][18][19]. This approach utilizes high-resolution Orbitrap mass spectrometry (MS) and differs from traditional single-cell approaches by determining the molecular signature of the secreted autoantibody proteome.…”

Section: Introductionmentioning

confidence: 99%

An immunodominant La/SSB autoantibody proteome derives from public clonotypes

Thurgood

Arentz

Lindop

et al. 2013

Clinical and Experimental Immunology

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SummaryThe La/SSB autoantigen is a major target of long-term humoral autoimmunity in primary Sjögren's Syndrome (SS) and systemic lupus erythematosus. A majority of patients with linked anti-Ro60/Ro52/La responses target an NH2-terminal epitope designated LaA that is expressed on Ro/La ribonucleoprotein complexes and the surface membrane of apoptotic cells. In this study, we used high-resolution Orbitrap mass spectrometry to determine the clonality, isotype and V-region sequences of LaA-specific autoantibodies in seven patients with primary SS. Anti-LaA immunoglobulin (Ig)Gs purified from polyclonal sera by epitope-specific affinity chromatography were analysed by combined database and de-novo mass spectrometric sequencing. Autoantibody responses comprised two heavily mutated IgG1 kappa-restricted monoclonal species that were shared (public) across unrelated patients; one clonotype was specified by an IGHV3-30 heavy chain paired with IGKV3-15 light chain and the second by an IGHV3-43/IGKV3-20 pairing. Shared amino acid replacement mutations were also seen within heavy and light chain complementarity-determining regions, consistent with a common breach of B cell tolerance followed by antigendriven clonal selection. The discovery of public clonotypic autoantibodies directed against an immunodominant epitope on La, taken together with recent findings for the linked Ro52 and Ro60 autoantigens, supports a model of systemic autoimmunity in which humoral responses against protein-RNA complexes are mediated by public sets of autoreactive B cell clonotypes.

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“…First, Scheid et al cloned 576 new human immunodeficiency virus (HIV) antibodies from four unrelated individuals and found that despite extensive hypermutation, these antibodies shared consensus sequences in both framework and CDR V regions of IgH chains (9). In another study on Sjö gren syndrome, secreted human Ro52 antibody from unrelated patients was found to share public V region sequences (27). Finally, our group observed 28 common Ig-derived sequences in paraneoplastic anti-Hu-syndrome that were specific for autoantigen and were found exclusively in samples from single autoantibody-defined clinical neurological entities (12).…”

Section: Discussionmentioning

confidence: 99%

Cerebrospinal-fluid-derived Immunoglobulin G of Different Multiple Sclerosis Patients Shares Mutated Sequences in Complementarity Determining Regions

Singh

Stoop

Stingl

et al. 2013

Molecular & Cellular Proteomics

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B lymphocytes play a pivotal role in multiple sclerosis pathology, possibly via both antibody-dependent and -independent pathways. Intrathecal immunoglobulin G in multiple sclerosis is produced by clonally expanded B-cell populations. Recent studies indicate that the complementarity determining regions of immunoglobulins specific for certain antigens are frequently shared between different individuals. In this study, our main objective was to identify specific proteomic profiles of mutated complementarity determining regions of immunoglobulin G present in multiple sclerosis patients but absent in healthy controls. To achieve this objective, we purified immunoglobulin G from the cerebrospinal fluid of 29 multiple sclerosis patients and 30 healthy controls and separated the corresponding heavy and light chains via SDS-PAGE. Subsequently, bands were excised, trypsinized, and measured with high-resolution mass spectrometry. We sequenced 841 heavy and 771 light chain variable region peptides. We observed 24 heavy and 26 light chain complementarity determining regions that were solely present in a number of multiple sclerosis patients. Using stringent criteria for the identification of common peptides, we found five complementarity determining regions shared in three or more patients and not in controls. Interestingly, one complementarity determining region with a single mutation was found in six patients. Additionally, one other patient carrying a similar complementarity determining region with another mutation was observed. In addition, we found a skew in the -to-ratio and in the usage of certain variable heavy regions that was previously observed at the transcriptome level. At the protein level, cerebrospinal fluid immunoglobulin G shares common characteristics in the antigen binding region among different multiple sclerosis patients. The indication of a shared fingerprint may indicate common antigens for B-cell activation. Molecular & Cellular Proteomics 12:

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Secreted human Ro52 autoantibody proteomes express a restricted set of public clonotypes

Cited by 34 publications

References 22 publications

Serum SmD autoantibody proteomes are clonally restricted and share variable-region peptides

Serum SmD autoantibody proteomes are clonally restricted and share variable-region peptides

An immunodominant La/SSB autoantibody proteome derives from public clonotypes

Cerebrospinal-fluid-derived Immunoglobulin G of Different Multiple Sclerosis Patients Shares Mutated Sequences in Complementarity Determining Regions

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