Monoclonal antibody FMC7 detects a conformational epitope on the CD20 molecule: evidence from phenotyping after rituxan therapy and transfectant cell analyses.
Numerous studies have reported that monoclonal antibody (mAb) FMC7 detects an antigen present on only a subset of circulating B lymphocytes. In particular, this mAb may distinguish typical B-cell chronic lymphocytic leukemia (FMC7 negative) from other types of B-cell non-Hodgkin lymphoma (B-NHL; FMC7 positive). We treated patients with B-NHL with Rituxan, a chimeric CD20 mAb, and observed abrogation of staining not only with prototype CD20 mAb B-1 but also with mAb FMC7. To investigate the relation between antigens CD20 and FMC7, we performed mutual blocking studies that showed mutual inhibition of FMC7 and CD20. In addition, FMC7 modulated CD23 expression and confirmed the presence of mAb B-1 in B-lymphoblastoid cell lines CESS and JVM. Transient transfection of myeloid cell line K562 with plasmid containing CD20-encoding cDNA produced de novo expressions of CD20 and FMC7. Our data indicate that FMC7 binds to a particular conformation of the CD20 antigen, probably to a multimeric CD20 complex. We assume that FMC7 stains positively only when CD20 antigen is present in high densities and in the postulated multimeric complex formation.
Serke
S
S
Department of Hematology and Oncology, Humboldt University, Berlin, Germany. serke@charite.de
Schwaner
I
I
Yordanova
M
M
Szczepek
A
A
Huhn
D
D
D019138:Antibodies, Blocking / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000627:therapeutic use*; D058846:Antibodies, Monoclonal, Murine-Derived; D000937:Antigen-Antibody Reactions; D018951:Antigens, CD20 / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis* / Q000378:metabolism; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D018274:Electroporation; D000939:Epitopes; D005434:Flow Cytometry; D006023:Glycoproteins; D006801:Humans; D016130:Immunophenotyping; D020014:K562 Cells; D016393:Lymphoma, B-Cell / Q000188:drug therapy* / Q000276:immunology; D010641:Phenotype; D000069283:Rituximab; D014162:Transfection
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D058846:Antibodies, Monoclonal, Murine-Derived; D018951:Antigens, CD20; D000944:Antigens, Differentiation, B-Lymphocyte; D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D000939:Epitopes; D006023:Glycoproteins; C112523:MS4A1 protein, human; D000069283:Rituximab