Close proximity, but not VLA-4-dependent adherence between pre-B cells and bone marrow stromal cells, is required for DMBA-induced apoptosis of pre-B cells in vitro

We have previously reported that 7,12-dimethylbenz[a]anthracene (DMBA) induced apoptosis in precursor B lymphocytes (pre-B cells) only when they were co-cultured with bone marrow stromal (BMS) cells. The goal of this research was to determine whether this process was dependent on the adherence of the pre-B cells and stromal cells. Conditioned media from DMBA-treated BMS cells induced apoptosis in pre-B cells, but only when the pre-B cells were co-cultured with stromal cells. This result suggested that stromal cells may release a soluble factor that initiates apoptosis, but their presence was still required for apoptosis. When the stromal cells and pre-B cells were separated with a membrane filter insert, DMBA-induced apoptosis of the pre-B cells was blocked suggesting that contact with or close proximity to stromal cells was required for apoptosis. The addition of an anti-VLA-4 Mab disrupted adherence of pre-B cells to the stromal cell monolayer, but did not diminish the numbers of apoptotic pre-B cells. The results of this study support the hypothesis stromal cells and pre-B cells must be in close proximity for apoptosis to occur, but direct interaction via VLA-4 and VCAM-1 is unlikely to be required for this response.