Persistent intrathecal secretion of oligoclonal, Borrelia burgdorferi-specific IgG in chronic meningoradiculomyelitis

Martin, Roland; Martens, Uwe M.; Sticht-Groh, V.; Dörries, Rüdiger; Krüger, H.

doi:10.1007/bf00314352

Cited by 58 publications

(19 citation statements)

References 25 publications

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“…No cross-reactivity of RA77 with T. pallidum spirochetes or E. coli could be detected, indicating that RA77 is specific for B. burgdorferi. This finding of a B. burgdorferi-specific antibody produced by a cePC is consistent with previous studies showing that OCB in the CSF of NB patients contain antibodies directed against B. burgdorferi (10,15), particularly in view of recent data from our laboratory supporting the conclusion that cePC are in fact the producers of OCB (unpublished data). To date, the only other inflammatory disease for which a link between CNS-derived plasma cells and the causative agent has been demonstrated is subacute sclerosing panencephalitis.…”

Section: Discussionsupporting

confidence: 92%

“…Analysis of immunoglobulins (Ig) in the CSF has revealed specificity for B. burgdorferi (10,15) and also for CNS autoantigens (12). However, the mechanisms underlying B. burgdorferi-directed humoral immune defense and CNS tissue damage still remain poorly understood, and more in-depth characterization of the focused antibody response in the CSF has been hampered by both the limited availability and the oligoclonal/polyclonal composition of CSF Ig.…”

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confidence: 99%

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Pathogen Specificity and Autoimmunity Are Distinct Features of Antigen-Driven Immune Responses in Neuroborreliosis

et al. 2007

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Neuroborreliosis (NB) is a chronic infectious disease of the central nervous system (CNS) caused by a tick-borne spirochete, Borrelia burgdorferi. In addition to direct effects of the causative infectious agent, additional immunity-mediated mechanisms are thought to play a role in the CNS pathology of NB. In order to further understand the involvement of humoral immune mechanisms in NB, we dissected the intrathecal antibody responses down to the single-plasma-cell level. Starting with single-cell reverse transcription-PCR of fluorescence-activated cell sorter-sorted cerebrospinal fluid plasma cells from an NB patient, we identified expanded clones and resurrected the antigen specificity of their secreted antibodies through recombinant expression of the correctly paired immunoglobulin heavy-and light-chain genes as monoclonal antibodies (MAbs). As expected, we found specificity for the causative infectious agent, B. burgdorferi, among the clonally expanded plasma cell (cePC)-derived MAbs. However, from an independent cePC of the same patient, we could derive MAbs specific for human CNS myelin, without detectable cross-reactivity with B. burgdorferi antigens. While reactivity against B. burgdorferi is a known feature of humoral immune responses in NB, we show (i) that immune responses specific for self antigens may be a distinct feature of CNS infections independent of pathogen reactivity and (ii) that humoral autoimmunity in NB (since found in cePC) is the result of a truly antigen-driven immune response. Our findings indicate that in NB mechanisms may be at play that induce distinct immune responses specific for pathogen and self antigens independent from "molecular mimicry

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Section: Discussionsupporting

confidence: 92%

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confidence: 99%

Pathogen Specificity and Autoimmunity Are Distinct Features of Antigen-Driven Immune Responses in Neuroborreliosis

et al. 2007

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“…This strategy revealed full-length cDNA sequences of matching Ig chains and allowed us to produce recombinant OCB (rOCB) antibodies. As proof of concept, we show that an OCB antibody from a patient with neuroborreliosis (NB) binds to a protein from Borrelia (25). Then, we characterized six rOCB antibodies from four MS patients and identified three different OCB target autoantigens.…”

Section: Significancementioning

confidence: 97%

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

Brändle

Obermeier

Şenel

et al. 2016

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

151

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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“…Studies in which the specificity of CSF OGBs was analyzed showed that the antibodies were directed against the agent that causes disease (1)(2)(3)(4)(5)(6)(7)(8). For example, the OGBs in SSPE CSF and brain are antibodies directed against MV (1).…”

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confidence: 99%

Laser-capture microdissection of plasma cells from subacute sclerosing panencephalitis brain reveals intrathecal disease-relevant antibodies

Burgoon

Keays

Owens

et al. 2005

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

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Increased IgG and oligoclonal bands (OGBs) are found in the cerebrospinal fluid (CSF) of humans with chronic infectious CNS diseases such as neurosyphilis, cryptococcal and tuberculous meningitis, Lyme disease, some viral meningitides, varicella zoster virus vasculopathy, and subacute sclerosing panencephalitis (SSPE), a chronic encephalitis caused by measles virus (MV). Studies in which the specificity of CSF OGBs was analyzed showed that the antibodies were directed against the agent that causes disease (1-8). For example, the OGBs in SSPE CSF and brain are antibodies directed against MV (1). These studies have led to the hypothesis that the oligoclonal IgG in the brain and CSF of patients with a chronic inflammatory CNS disease of unknown etiology, such as multiple sclerosis (MS), sarcoidosis, and Behcet's disease, is antibody directed against the agent that causes disease. Although OGBs are found in 88-100% of CSF from MS patients (9), the corresponding antigens remain unknown (10, 11). Such unanswered questions point to the need for improved techniques to identify disease-relevant antibodies and their cognate antigens and suggest the promise of such techniques in revealing the causes of inflammatory diseases with unknown etiologies.Here, we used laser-capture microdissection (LCM) to isolate individual CD38ϩ plasma cells from the brain of a patient with SSPE. Single-cell RT-PCR was used to amplify individual IgG heavy (H) and light (L) chain sequences expressed by each cell. Based on overrepresented Ig sequences, we constructed functional recombinant antibodies and identified their target antigens.Materials and Methods SSPE Brain. Brain removed from a 14-year-old male SSPE patient 5 h after death was flash-frozen and stored at Ϫ70°C. Sections (7 m) of frozen SSPE brain were prepared on nonplus glass slides (Fisher Scientific) at Ϫ30°C, fixed in acetone for 5 min at Ϫ20°C, and immunostained at 0°C for CD38ϩ cells. Briefly, sections were treated with 0.1% H 2 O 2 for 30 sec, incubated for 2 min with PBS containing 10% goat serum and then with a 1:50 dilution of mouse anti-human CD38 antibody (DakoCytomation, DAKO) for 10 min, rinsed in PBS, and incubated for 5 min with a 1:100 dilution of horseradish peroxidase-conjugated horse anti-mouse antibody (Vector Laboratories) and 5% goat serum in PBS. After rinsing in PBS, sections were incubated for 5 min with DAB substrate (Vector Laboratories), counterstained with hematoxylin, dehydrated in nuclease-free graded alcohols to 100% ethanol, and cleared with two rinses of H 2 0-free xylene. To preserve RNA, all aqueous solutions contained 200 units͞ml RNase inhibitor (Fisher Scientific), and the total time of tissue exposure to aqueous solutions was Ͻ25 min at 0°C. Staining was visualized by light microscopy.LCM and RT-PCR. LCM was performed on a PixCell IIe microscope (Arcturus Engineering, Mountain View, CA) with CapSure HS caps by using a pulse power of 70 mW, a 7.5-m laser spot diameter, pulse duration of 5 ms, and target voltage of 170 mV. Individual CD38ϩ cells ...

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Persistent intrathecal secretion of oligoclonal, Borrelia burgdorferi-specific IgG in chronic meningoradiculomyelitis

Cited by 58 publications

References 25 publications

Pathogen Specificity and Autoimmunity Are Distinct Features of Antigen-Driven Immune Responses in Neuroborreliosis

Pathogen Specificity and Autoimmunity Are Distinct Features of Antigen-Driven Immune Responses in Neuroborreliosis

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

Laser-capture microdissection of plasma cells from subacute sclerosing panencephalitis brain reveals intrathecal disease-relevant antibodies

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