Abstract
We describe a method for correlating the immunoglobulin (Ig) proteomes with the B cell transcriptomes in human fluid and tissue samples, using multiple sclerosis as a paradigm. Oligoclonal Ig bands and elevated numbers of clonally expanded B cells in the cerebrospinal fluid (CSF) are diagnostic hallmarks of multiple sclerosis. Here we compared the Ig transcriptomes of B cells with the corresponding Ig proteomes in CSF samples from four subjects with multiple sclerosis. We created individual Ig transcriptome databases that contained the subject-specific mutations introduced by V(D)J recombination and somatic hypermutation and then searched the CSF for corresponding characteristic peptides by mass spectrometry. In each sample, the Ig transcriptomes and proteomes strongly overlapped, showing that CSF B cells indeed produce the oligoclonal Ig bands. This approach can be applied to other organ-specific diagnostic fluid or tissue samples to compare the Ig transcripts of local B cells with the corresponding antibody proteomes of individual subjects.
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References
Kabat, E.A., Moore, D.H. & Landow, H. An electrophoretic study of the protein components in cerebrospinal fluid and their relationship to the serum proteins. J. Clin. Invest. 21, 571–577 (1942).
Qin, Y., Duquette, P., Zhang, Y., Poole, R. & Antel, J.P. Clonal expansion and somatic hypermutation of VH genes of B cells from cerebrospinal fluid in multiple sclerosis. J. Clin. Invest. 102, 1045–1050 (1998).
Owens, G.P. et al. Restricted use of VH4 germline segments in an acute multiple sclerosis brain. Ann. Neurol. 43, 236–243 (1998).
Baranzini, S.E. et al. B cell repertoire diversity and clonal expansion in multiple sclerosis brain lesions. J. Immunol. 163, 5133–5144 (1999).
Colombo, M. et al. Accumulation of clonally related B lymphocytes in the cerebrospinal fluid of multiple sclerosis patients. J. Immunol. 164, 2782–2789 (2000).
Owens, G.P. et al. Single-cell repertoire analysis demonstrates that clonal expansion is a prominent feature of the B cell response in multiple sclerosis cerebrospinal fluid. J. Immunol. 171, 2725–2733 (2003).
Monson, N.L. et al. Receptor revision and atypical mutational characteristics in clonally expanded B cells from the cerebrospinal fluid of recently diagnosed multiple sclerosis patients. J. Neuroimmunol. 158, 170–181 (2005).
Odegard, V.H. & Schatz, D.G. Targeting of somatic hypermutation. Nat. Rev. Immunol. 6, 573–583 (2006).
Vandvik, B., Natvig, J.B. & Wiger, D. IgG1 subclass restriction of oligoclonal IgG from cerebrospinal fluids and brain extracts in patients with multiple sclerosis and subacute encephalitides. Scand. J. Immunol. 5, 427–436 (1976).
Kaschka, W.P., Theilkaes, L., Eickhoff, K. & Skvaril, F. Disproportionate elevation of the immunoglobulin G1 concentration in cerebrospinal fluids of patients with multiple sclerosis. Infect. Immun. 26, 933–941 (1979).
Losy, J., Mehta, P.D. & Wisniewski, H.M. Identification of IgG subclasses' oligoclonal bands in multiple sclerosis CSF. Acta Neurol. Scand. 82, 4–8 (1990).
Link, H. & Zettervall, O. Multiple sclerosis: disturbed κ:λ chain ratio of immunoglobulin G in cerebrospinal fluid. Clin. Exp. Immunol. 6, 435–438 (1970).
Qian, W.J., Jacobs, J.M., Liu, T. & Camp, D.G. 2nd & Smith, R.D. Advances and challenges in liquid chromatography–mass spectrometry–based proteomics profiling for clinical applications. Mol. Cell. Proteomics 5, 1727–1744 (2006).
Warren, K.G., Catz, I. & Steinman, L. Fine specificity of the antibody response to myelin basic protein in the central nervous system in multiple sclerosis: the minimal B cell epitope and a model of its features. Proc. Natl. Acad. Sci. USA 92, 11061–11065 (1995).
Genain, C.P., Cannella, B., Hauser, S.L. & Raine, C.S. Identification of autoantibodies associated with myelin damage in multiple sclerosis. Nat. Med. 5, 170–175 (1999).
O'Connor, K.C. et al. Antibodies from inflamed central nervous system tissue recognize myelin oligodendrocyte glycoprotein. J. Immunol. 175, 1974–1982 (2005).
Owens, G.P. et al. VH4 gene segments dominate the intrathecal humoral immune response in multiple sclerosis. J. Immunol. 179, 6343–6351 (2007).
Serafini, B., Rosicarelli, B., Magliozzi, R., Stigliano, E. & Aloisi, F. Detection of ectopic B cell follicles with germinal centers in the meninges of patients with secondary progressive multiple sclerosis. Brain Pathol. 14, 164–174 (2004).
Meinl, E., Krumbholz, M. & Hohlfeld, R. B lineage cells in the inflammatory CNS environment: migration, maintenance, local antibody production and therapeutic modulation. Ann. Neurol. 59, 880–892 (2006).
Polman, C.H. et al. Diagnostic criteria for multiple sclerosis: 2005 Revisions to the “McDonald Criteria”. Ann. Neurol. 58, 840–846 (2005).
Wardemann, H. et al. Predominant autoantibody production by early human B cell precursors. Science 301, 1374–1377 (2003).
Shevchenko, A., Wilm, M., Vorm, O. & Mann, M. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 68, 850–858 (1996).
Zverlov, V.V., Kellermann, J. & Schwarz, W.H. Functional subgenomics of Clostridium thermocellum cellulosomal genes: identification of the major catalytic components in the extracellular complex and detection of three new enzymes. Proteomics 5, 3646–3653 (2005).
Kabat, E.A. & Wu, T.T. Identical V region amino acid sequences and segments of sequences in antibodies of different specificities. Relative contributions of VH and VL genes, minigenes, and complementarity-determining regions to binding of antibody-combining sites. J. Immunol. 147, 1709–1719 (1991).
Obermeier, B., Mentele, R., Malotka, J. & Dornmair, K. Analysis of patient-specific immunoglobulin proteomes and transcriptomes by PCR cloning and mass spectrometry. Nat. Protoc. published online, doi: 10.1038/nprot.2008.87 (18 May 2008).
Acknowledgements
This work was supported by the Deutsche Forschungsgemeinschaft grant SFB 571-A1 and the Hermann and Lilly Schilling Foundation. We thank M. Wick and M. Krumbholz for valuable support and J. Benson for editing the manuscript.
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B.O. conducted most of the experiments and prepared the figures, R.M. and J.K. participated in IEF and mass spectrometry experiments, J.M. participated in cloning of Ig chains, T.K. provided CSF samples and clinical data, H.W. and F.L. supervised the project and participated in writing the manuscript and K.D. and R.H. initiated and supervised the project and wrote the manuscript.
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Obermeier, B., Mentele, R., Malotka, J. et al. Matching of oligoclonal immunoglobulin transcriptomes and proteomes of cerebrospinal fluid in multiple sclerosis. Nat Med 14, 688–693 (2008). https://doi.org/10.1038/nm1714
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DOI: https://doi.org/10.1038/nm1714
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