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American Journal of Pathology, Vol 136, 1215-1222, Copyright © 1990 by American Society for Investigative Pathology

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Antibody L26 recognizes an intracellular epitope on the B-cell- associated CD20 antigen

DY Mason, WM Comans-Bitter, JL Cordell, MA Verhoeven and JJ van Dongen
Department of Haematology, John Radcliffe Hospital, Oxford, England.

Monoclonal antibody L26 is a highly selective marker of B cells and B- cell neoplasms in paraffin-embedded tissues, but it suffers from the drawback that the target molecule has not been identified. In this paper we provide evidence by two independent techniques that antibody L26 recognizes an intracellular epitope on the CD20 antigen (a pan B- cell marker). When this antigen was redistributed on the surface of unfixed viable B cells by incubation with monoclonal anti-CD20 followed by anti-mouse Ig, the diffuse cytoplasmic staining of L26 was abolished and replaced by coincident dotlike labeling for antibody L26 and the CD20 antigen. None of the other antibodies tested (covering 10 different B-cell-associated antigens) had this effect on the L26 staining pattern. Furthermore, COS-1 cells transfected with cDNA encoding the CD20 molecule gave positive staining with antibody L26 and with two other CD20 reagents, but not with antibodies to other pan B- cell markers (eg, CD19 and CD22).

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