Detection of Antigen Specific B-Cells in Tissues
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 WordsAutoantigens autoantibodies B-cells plasma cells immunohistochemistry immunofluorescence antibody synthesis antibody subclass biotinylation