CD22 antigen: biosynthesis, glycosylation and surface expression of a B lymphocyte protein involved in B cell activation and adhesion.
The CD22 antigen, although present in the cytoplasm of early and immature B cells, first appears on the cell surface of mature B lymphocytes. The phenotypic patterns and some functional properties of the surface-expressed CD22 antigen are well described, but little is known about its molecular structure. We have therefore investigated the relationship of the two CD22 glycoproteins (140/130 kd), and the influence of their complex N-glycosylation on surface expression and antibody recognition. Comparative peptide mapping of the 100 and 80 kd protein cores, obtained by endoglycosidase F treatment, revealed a common structure shared by both protein cores. In pulse-chase experiments the mature glycoproteins originated from two separate precursor molecules, indicating that the two proteins may be generated by different RNA processing. In cell lysates a CD22 specific polyclonal anti-serum recognized both molecules the size of the protein cores and glycosylated CD22 molecules, whereas in membrane preparations only the glycosylated forms were detected. The CD22 mAb HD39 reacted exclusively with the glycoprotein froms in either cellular preparation. The influence of glycosylation on surface expression and epitope recognition was investigated in more detail by applying various inhibitors of the glycosylation pathway. 1-Deoxymannojirimycin and Swainsonine, which block glycosylation at the high-mannose and hybrid-type stages respectively, modulated the CD22 antigen but did not alter its surface expression. Tunicamycin blocked de novo glycosylation and led to reduced surface recognition of the CD22 antigen. Together, these results suggested that comparatively simple oligosaccharide structures of high-mannose type are sufficient for surface expression of the CD22 antigen and for epitope recognition by mAb HD39. It is most likely that glycosylation is required to stabilize epitopes in the protein moiety recognized by CD22 mAb. Finally, we demonstrated the presence of glycosylated, cytoplasmic CD22 antigen in CD22 surface negative B-lineage ALL cells. This finding led us to conclude that complex glycosylation does not provide the determining signal for the switch from cytoplasmic to surface expression of the CD22 antigen.
Schwartz-Albiez
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R
Institute of Immunology and Genetics, German Cancer Research Center, Heidelberg.
Dörken
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B
Monner
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DA
Moldenhauer
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G
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000096:biosynthesis* / Q000378:metabolism; D000944:Antigens, Differentiation, B-Lymphocyte / Q000096:biosynthesis* / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000276:immunology; D002448:Cell Adhesion / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000096:biosynthesis / Q000378:metabolism; D000939:Epitopes; D006031:Glycosylation; D006801:Humans; D037102:Lectins; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D051918:Sialic Acid Binding Ig-like Lectin 2; D014407:Tumor Cells, Cultured / Q000276:immunology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C497867:CD22 protein, human; D002241:Carbohydrates; D015815:Cell Adhesion Molecules; D000939:Epitopes; D037102:Lectins; D051918:Sialic Acid Binding Ig-like Lectin 2