Phytohemagglutinin (PHA)-resistant Chinese hamster ovary cell mutants acquire sensitivity to the lectin after fusion with liposomes containing PHA receptor glycoproteins.
Phytohemagglutinin receptor glycoproteins have been inserted into phospholipid vesicles and these have been fused with phytohemagglutinin-resistant chinese hamster ovary cells. Our results show that the fused cells acquire "neoreceptors" for the lectin phytohemagglutinin. Fluorescence activated cell sorting analyses show that approximately 40% of the cells fused with the receptor-containing vesicles. Studies with 125I-labelled lectin showed that fused cells bound three times more ligand than untreated mutant cells. Furthermore, lectin receptors were functionally inserted in the mutant cell plasma membrane. Fused cells cultured in the presence of lectin (200 micrograms ml-1) lost rapidly (8 hours or less) their ability to incorporate [3H] thymidine. Whereas mutant cells cultured for 16 hours in the presence of 50-400 micrograms ml-1 of lectin remained viable, fused cells showed a 45% decrease in 3H-labelled nucleotide incorporation. The method described here should be of general applicability for the study of lectin-dependent cytotoxicity in chinese hamster ovary cell lines.
D000818:Animals; D002459:Cell Fusion; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004351:Drug Resistance; D005260:Female; D006023:Glycoproteins / Q000032:analysis*; D008081:Liposomes; D009154:Mutation; D010053:Ovary; D010835:Phytohemagglutinins / Q000494:pharmacology*; D011975:Receptors, Mitogen / Q000032:analysis*; D013936:Thymidine / Q000378:metabolism
D006023:Glycoproteins; D008081:Liposomes; D010835:Phytohemagglutinins; D011975:Receptors, Mitogen; D013936:Thymidine