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Detection of intrathecal immunoglobulin G synthesis by capillary isoelectric focusing immunoassay in oligoclonal band negative multiple sclerosis

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Abstract

Oligoclonal immunoglobulin G bands (OCBs) restricted to the cerebrospinal fluid indicate intrathecal inflammation. Using isoelectric focusing and immunoblotting, they are detected in about 95 % of patients with clinically definite multiple sclerosis (MS). To elucidate whether in the remaining 5 % OCBs are truly absent or alternatively missed due to insufficient sensitivity of the routine measurement, we employed a new, highly sensitive nanoscale method for OCB detection. Capillary isoelectric focusing followed by immunological detection served to analyze OCBs in 33 well-characterized OCB-negative and 10 OCB-positive MS patients as well as in 100 OCB-negative control patients with non-inflammatory neurological diseases and 30 OCB-positive control patients with inflammatory neurological diseases. We detected intrathecal immunoglobulin G production in 10 out of 33 MS patients (30 %), initially diagnosed as being OCB-negative, and in all 10 OCB-positive MS patients, but in only 3 out of 100 non-inflammatory neurological controls (3 %) and in 29 of 30 inflammatory neurological controls (97 %). At least about one-third of MS patients without intrathecal immunoglobulin G synthesis according to standard methods are OCB-positive. Advanced methods for OCB detection may increase the analytical sensitivity for detecting OCB in patients with MS who are OCB-negative according to current routine methods.

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Acknowledgments

This work was supported by BMBF (German Federal Ministry for Education and Research): Kompetenznetz Multiple Sklerose (KKNMS), Kompetenznetz Neurodegenerative Demenzen (KNDD, FTLDc), and JPND projects: BIOMARKAPD, SOPHIA, FrontALS), Foundation of the State Baden-Wuerttemberg and the EU (NADINE) (Contract No. 246513). The work of MB, IB and AW was supported by a grant from Novartis for multiple sclerosis biomarker research. We especially thank our patients for participating in these studies and R. Aksamija for expert technical assistance.

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    Authors and Affiliations

    1. Department of Neurology, University of Ulm, Oberer Eselsberg 45, 89081, Ulm, Germany

      Steffen Halbgebauer, André Huss, Petra Steinacker, Patrick Oeckl, Hayrettin Tumani & Markus Otto

    2. Department of Neurology, University of Würzburg, Josef-Schneider-Str. 11, 97080, Würzburg, Germany

      Mathias Buttmann, Isabel Brecht & Andreas Weishaupt

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    1. Steffen Halbgebauer
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    Correspondence to Markus Otto.

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    All patients gave written informed consent for the use of remaining samples for research. This procedure and sample analysis for the purpose of this study was approved by the local Ethics Committees in Ulm and Würzburg in accordance with the Declaration of Helsinki.

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    S. Halbgebauer and A. Huss contributed equally.

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    Halbgebauer, S., Huss, A., Buttmann, M. et al. Detection of intrathecal immunoglobulin G synthesis by capillary isoelectric focusing immunoassay in oligoclonal band negative multiple sclerosis. J Neurol 263, 954–960 (2016). https://doi.org/10.1007/s00415-016-8094-3

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    • DOI: https://doi.org/10.1007/s00415-016-8094-3

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