CVIC Lab.CardioVascular-Ion Channel Laboratory

Regulation of B cell receptor (BCR)-induced Ca²⁺ signaling by CD40 co-stimulation was compared in long-term BCR-stimulated immature (WEHI-231) and mature (Bal-17) B cells. In response to long-term pre-stimulation of immature WEHI-231 cells to α-IgM antibody (0.5 ∼ 48 hr), the initial transient decrease in BCR-induced [Ca²⁺]_i was followed by spontaneous recovery to control level within 24 hr. The recovery of Ca ²⁺ signaling in WEHI-231 cells was not due to restoration of internalized receptor but instead to an increase in the levels of PLCγ2 and IP₃R-3. CD40 co-stimulation of WEHI-231 cells prevented BCR-induced cell cycle arrest and apoptosis, and it strongly inhibited the recovery of BCR-induced Ca²⁺ signaling. CD40 co-stimulation also enhanced BCR internalization and reduced expression of PLCγ2 and IP ₃R-3. Pre-treatment of WEHI-231 cells with the antioxidant N-acetyl-L-cysteine (NAC) strongly inhibited CD40-mediated prevention of the recovery of Ca²⁺ signaling. In contrast to immature WEHI-231 cells, identical long-term α-IgM pre-stimulation of mature Bal-17 cells abolished the increase in BCR-induced [Ca²⁺]_i, regardless of CD40 co-stimulation. These results suggest that CD40-mediated signaling prevents antigen-induced cell cycle arrest and apoptosis of immature B cells through inhibition of sustained BCR-induced Ca²⁺ signaling.