Detection of Antigen Specific B-Cells in Tissues

  • Marie Wahren-Herlenius
  • Stina Salomonsson
Part of the Methods in Molecular Medicine book series (MIMM, volume 136)


The role of B-cells and autoantibodies in tissue destructive events of autoimmune diseases is emerging, and thereby increasing interest in identifying the presence and location of autoreactive B-cells and autoantibody secreting plasma cells. For visualization and analysis of the autoreactive B-cells, the antigen of interest is selected and produced and purified from native or recombinant sources. Biotinylation of the purified antigen and subsequent use in immunohistochemistry with sections from tissue under analysis permits detection of the autoreactive B-cells and plasma cells. Double staining with cell-specific markers or for the presence of intracellular Ig allows characterization of the cell or determination of Ig isotype of the autoantibody produced by the individual autoreactive B -cell, leading to better understanding of the role of the particular antibody in the inflammatory cascade of the organ. With this technique, identification as well as quantification of autoreactive cells within tissues may be performed; it is also possible to analyze the spatial relation to residual cells or other infiltrating cells of the target organ.

Key Words

Autoantigens autoantibodies B-cells plasma cells immunohistochemistry immunofluorescence antibody synthesis antibody subclass biotinylation 


  1. 1.
    Chan, O. T., Madaio, M. P., and Shlomchik, M. J. (1999) The central and multiple roles of B cells in lupus pathogensis. Immunol. Rev. 169, 107–121.PubMedCrossRefGoogle Scholar
  2. 2.
    Davidson, A., and Diamond, B. (2001) Autoimmune diseases. N. Engl. J. Med. 345, 340–350.PubMedCrossRefGoogle Scholar
  3. 3.
    Tengnér, P., Halse, A.-K., Haga, H.-J., Jonsson, R., and Wahren-Herlenius, M. (1998) Detection of anti-Ro/SSA and anti-La/SSB autoantibody-producing cells in salivary glands from patients with Sjögren’s syndrome. Arthritis Rheum 41, 2238–2248.PubMedCrossRefGoogle Scholar
  4. 4.
    Salomonsson, S., and Wahren-Herlenius, M. (2003) Local production of Ro/SSA and La/SSB autoantibodies in the target organ coincides with high levels of circulating antibodies in sera of patients with Sjogren’s syndrome. Scand J Rheumatol 32(2), 79–82.PubMedCrossRefGoogle Scholar
  5. 5.
    Salomonsson, S., Jonsson, M. V., Skarstein, K., et al. (2003) Cellular basis of ectopic germinal center formation and autoantibody production in the target organ of patients with Sjögren’s syndrome. Arthritis Rheum 48, 3187–3201.PubMedCrossRefGoogle Scholar
  6. 6.
    Salomonsson, S., Larsson, P., Tengnér, P., et al. (2002) Expression of the B cellattracting chemokine CXCL13 in the target organ and autoantibody production in ectopic lymphoid tissue in the chronic inflammatory disease Sjögren’s syndrome. Scand J. Immunol 55, 336–342.PubMedCrossRefGoogle Scholar
  7. 7.
    Becker, J. M., and Wilchek, M. (1972) Inactivation by avidin of biotin-modified bacteriophage. Biochim. Biophys. Acta 264, 165–170.PubMedGoogle Scholar
  8. 8.
    Heitzmann, H., and Richards, F. M. (1974) Use of the avidin-biotin complex for specific staining of biological membranes in electron microscopy. Proc. Natl. Acad. Sci. USA 71, 3537–3541.PubMedCrossRefGoogle Scholar
  9. 9.
    Heggeness, M. H., and Ash, J. F. (1977) Use of the avidin-biotin complex for the localization of actin and myosin with fluorescence microscopy. J. Cell Biol. 73, 783–788.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2007

Authors and Affiliations

  • Marie Wahren-Herlenius
    • 1
  • Stina Salomonsson
    • 1
  1. 1.Rheumatology Unit, Department of MedicineKarolinska HospitalStockholmSweden

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