Membrane IgM cross-linking is not coupled to protein kinase C translocation in WEHI-231 B lymphoma cells

Abstract

The early molecular events involved in the process of signal transduction via membrane immunoglobulins (mIg) include phosphatidyl inositol metabolism, intracellular Ca²⁺ mobilization, and protein kinase C (PKC) activation. Anti‐mIg antibodies exert either stimulating or inhibitory effects depending on the activation state and/or the differentiation stage of B cells. WEHI‐231 is a murine B lymphoma that becomes inactivated upon anti‐mIg treatment. This lymphoma has an immature B cell phenotype and is considered as a model for tolerance induction in B lymphocytes. In this study, we have investigated the relationship between mIg triggering, Ca²⁺ elevation, PKC translocation, and growth inhibition in WEHI‐231 cells. Monoclonal antibodies to μ and χ chains of the mIgM receptor promoted a rapid increase in intracytoplasmic Ca²⁺ and were potent inhibitors of cell growth. Ca²⁺ elevation and PKC translocation have been previously shown to be associated in B lymphocytes. To study the subcellular distribution of PKC in WEHI‐231 cells, we used enzymatic assays and immunodetection methods. Although phorbol 12‐myristate 13‐acetate induced a rapid and almost complete redistribution of cytosolic PKC to the membrane fraction, anti‐mIg treatment failed to modify the compartmentalization of PKC. These findings extend recent observations suggesting that B cell triggering through mIg receptors may involve additional pathways independent from PKC activation. PKC activation in normal B cells is also believed to provide a regulatory signal which limits the magnitude of the early signals produced by anti‐mIg. Such a regulatory control is unlikely in WEHI‐231 cells, due to the dissociation between Ca²⁺ mobilization and PKC translocation. Our findings therefore suggest that the sensitivity of immature B cells such as WEHI‐231 to the inhibitory effects of anti‐mIg antibodies may result in part from alterations of the phosphoinositide signal transduction pathway.