Related B cell clones that populate the CSF and CNS of patients with multiple sclerosis produce CSF immunoglobulin

doi:10.1016/j.jneuroim.2011.01.010

Journal of Neuroimmunology

Volume 233, Issues 1–2, April 2011, Pages 245-248

https://doi.org/10.1016/j.jneuroim.2011.01.010 Get rights and content

Abstract

We investigated the overlap shared between the immunoglobulin (Ig) proteome of the cerebrospinal fluid (CSF) and the B cell Ig-transcriptome of CSF and the central nervous system (CNS) tissue of three patients with multiple sclerosis. We determined the IgG-proteomes of CSF by mass spectrometry, and compared them to the IgG-transcriptomes from CSF and brain lesions, which were analyzed by cDNA cloning. Characteristic peptides that were identified in the CSF-proteome could also be detected in the transcriptomes of both, brain lesions and CSF, providing evidence for a strong overlap of the IgG repertoires in brain lesions and in the CSF.

Introduction

The role of B cells and antibodies in the pathogenesis of multiple sclerosis (MS) has become more appreciated (Hafler et al., 2005, Meinl et al., 2006). B cells and their products, antibodies, are detected in all compartments of the central nervous system (CNS): in the parenchyma (Ozawa et al., 1994), in the meninges, where they may form germinal-center-like structures (Serafini et al., 2004), but also in the cerebrospinal fluid (CSF), where oligoclonal B cell expansions and expanded antibody populations, so-called “oligoclonal bands” (OCBs), are observed (Baranzini et al., 1999, Colombo et al., 2000, Monson et al., 2005, Obermeier et al., 2008, Owens et al., 1998, Owens et al., 2009, Qin et al., 1998). We recently showed that the repertoires of IgG-proteins and of B cells in the CSF overlap to a great extent. This provided evidence that the CSF-resident B cells may contribute to the production of OCB-antibodies (Obermeier et al., 2008). However, it is not clear if CSF B cells are the exclusive source of OCB antibodies as we recently learned that the MS CNS harbors a B cell network of clonally related B cells that populate both the CSF and distinct regions of the CNS (Lovato et al., 2011).

We here asked the question to what extent the CSF IgG-protein repertoire, i.e. the CSF-proteome, is derived from the B cell receptor repertoires, i.e. the transcriptomes, of CSF and of parenchymal brain lesions. The starting point of this analysis therefore was the analysis of the IgG-protein repertoire from CSF by mass spectrometry. We then compared this repertoire to the IgG-transcriptome repertoires from CSF and CNS tissue, which we determined in parallel experiments by cDNA cloning. These data demonstrate that B cell clones are often shared between the CSF and CNS tissue and that these clones produce immunoglobulins present in the CSF.

Section snippets

Clinical samples

CNS tissue was collected at autopsy from two patients with clinically defined MS (MS-4 and MS-B2A) at the Department of Pathology at Brigham and Women's Hospital. White matter lesions (plaques) were macroscopically identified, dissected, and immediately snap-frozen. Because of initial diagnostic uncertainty a brain biopsy was performed on patient L-296 revealing an inflammatory demyelinating process consistent with MS. The CSF of MS-4 and MS-B2A was removed post-mortem and centrifuged. The CSF

Results

To analyze the IgG-proteome from CSF, we separated OCBs from CSF of three MS patients by non-reducing two dimensional gel-electrophoresis, isolated Heavy and Light IgG-chains (IgG-H and IgG-L) from the gels (Supplementary Fig. 1) and subjected them to MALDI and ESI mass spectrometry. For more than 50% of the peptides we could verify the sequences by tandem mass spectrometry. We then aligned the peptides to patient-specific IgG-transcript databases that were obtained independently from CSF and

Discussion

We found strong overlaps of the CSF IgG-H and Ig-L-chain proteomes with the Ig-transcriptomes from CSF and from brain lesions. This means that the antibody repertoire present in the CSF reflects Ig-transcripts of B cells populating both compartments. Our data therefore confirm our previous study where we showed a strong overlap between CSF-proteome and CSF-transcriptome (Obermeier et al., 2008), providing evidence that CSF-resident B cells produce CSF-resident antibodies. Here we extend this

Acknowledgments

We thank Ingrid Eiglmeier for expert technical assistance and Edgar Meinl and Gurumoorthy Krishnamoorthy for comments on the manuscript. This work was supported by grants from Deutsche Forschungsgemeinschaft (SFB 571-A1, KD and RH), the German Ministry for Education and Research (BMBF, “German Competence Network Multiple Sclerosis” (KKNMS), RH), and NeuroproMiSe (LSHM-CT-2005-018637-WPM5, KD, HW and WB). L.L. was supported by a training research fellowship FISM — Fondazione Italiana Sclerosi

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Short communicationRelated B cell clones that populate the CSF and CNS of patients with multiple sclerosis produce CSF immunoglobulin

Abstract

Introduction

Section snippets

Clinical samples

Results

Discussion

Acknowledgments

Anal. Biochem.

J. Neuroimmunol.

J. Neurol. Sci.

B cell repertoire diversity and clonal expansion in multiple sclerosis brain lesions

J. Immunol.

Accumulation of clonally related B lymphocytes in the cerebrospinal fluid of multiple sclerosis patients

J. Immunol.

Multiple sclerosis

Immunol. Rev.

Multiple sclerosis: T-cell receptor expression in distinct brain regions

Brain

Lipid microarrays identify key mediators of autoimmune brain inflammation

Nat. Med.

Receptor revision and atypical mutational characteristics in clonally expanded B cells from the cerebrospinal fluid of recently diagnosed multiple sclerosis

Brain

B lineage cells in the inflammatory CNS environment: migration, maintenance, local antibody production and therapeutic modulation

Ann. Neurol.

Short communication
Related B cell clones that populate the CSF and CNS of patients with multiple sclerosis produce CSF immunoglobulin