Inhibition of homotypic aggregation of a human Burkitt-lymphoma cell line.
The molecules involved in homotypic aggregation of the human Burkitt-lymphoma cell line Raji were investigated by inhibition of reaggregation with carbohydrates and glycoconjugates, by inhibition of glycosylation, and enzyme treatment of the cell surface. Complete inhibition of reaggregation was achieved with bovine submaxillary mucin. Asialomucin, on the other hand, was not effective in this assay. Another potent inhibitor of reaggregation was the ganglioside GMI. The common carbohydrate structure of these molecules is NeuNAc-(gal)-galNAc. Lactosamine, fucosyllactosamine, sialyllactosamine, complex mannose type, or Thomsen-Friedenreich antigen sequences are not involved in aggregation. Neuraminidase and chloroquine also abolished agglutination of cells. The finding that mucin, but not asialomucin, inhibits the reaction, demonstrates the importance of sialic acid in this process. Homotypic aggregation was shown to be resistant to trypsin. Using the glycosylation inhibitor tunicamycin we show that N-glycosidically linked carbohydrate chains are involved in aggregation. Swainsonine or castanospermine, which inhibit processing of terminal sialyllactosamines to the mannose core, did not interfere with the reaction supporting the results of the inhibition assay. The data presented suggest the involvement of 2 molecules in homotypic aggregation of human Burkitt-lymphoma cells. One component is a lectin-like molecule containing N-linked carbohydrate chains. The other component carries the neuraminidase-sensitive and trypsin-resistant determinant NeuNAc-(gal)-galNAc and, therefore, appears to be a glycolipid. This proposed lectin-carbohydrate interaction in homotypic aggregation is further supported by the frequently observed dependence of lectins on divalent cations as indicated by inhibition of aggregation with EDTA and EGTA.
Department of Hematology, Oncology and Immunology, Robert-Bosch-Krankenhaus, Stuttgart, FRG.
M F
MF
Wolf
U
U
Koerner
B
B
Klumpp
K
K
Schumacher
D002051:Burkitt Lymphoma / Q000473:pathology*; D002241:Carbohydrates / Q000494:pharmacology; D002449:Cell Aggregation / Q000187:drug effects*; D003864:Depression, Chemical; D004798:Enzymes / Q000494:pharmacology*; D006001:Glycoconjugates / Q000494:pharmacology*; D006031:Glycosylation; D006801:Humans; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology*
D002241:Carbohydrates; D004798:Enzymes; D006001:Glycoconjugates