Biochemical characterisation of leukaemia-associated antigen p24 defined by the monoclonal antibody BA-2

The leukaemia-associated cell surface antigen p24/BA-2 is a single polypeptide chain with a molecular weight of 24,000. Treatment with glycosidases or exposure of cells to tunicamycin failed to show any change in the molecular weight of the antigen when examined by SDS-polyacrylamide gel electrophoresis. In addition, it failed to bind to lectin affinity columns of concanavalin A, lentil lectin or ricinus communis lectin. This is consistent with the absence of N-asparagine linked oligosaccharide chains on the antigen. Pulse-chase labelling of protein p24 shows a post-translational modification resulting in a molecular weight increase of approx. 500-1000. Alkaline treatment resulted in a decrease in molecular weight of approximately the same amount, suggesting that p24 contain some O-glycosidically linked oligosaccharide. Protein p24 has a basic pI of 7.3 which is unchanged after neuraminidase treatment. Protein P24/Ba-2 cannot be labelled by either the lipophilic photoactivatable nitrene reagent, hexanoyldiiodo-N-(4-azido-2-nitrophenyl)tyramine, or with [32P]phosphate. This suggests that the molecule is non-integral in nature and that it does not form an intimate association with the lipid matrix. Identical molecular weights, when reduced and non-reduced antigens were compared, suggest that it contains no internal disulphide linkages and failure to detect any other band on gradient gel SDS-polyacrylamide gel electrophoresis from 5-15% suggests that is is not strongly associated with any other structure.     

Lymphokine-induced changes in macrophage transmembrane electrical potentials

Intracellular electrical potentials have been measured in nonactivated and immunologically-activated macrophages obtained from the peritoneal cavities of mice. Normal macrophage potentials were established and found to become significantly more electronegative after in vitro exposure for 10 minutes to a lymphokine-containing supernatant which induced macrophage activation. This approach may reflect very early concomitants of such activation and is also useful in the study of other immunologic systems.