Effect of cell density on in vitro mouse immunoglobulin E production.

In vitro immunoglobulin E (IgE) production was found to be sensitive to increasing cell concentration in culture wells. While class switching to IgE is intact as suggested by surface IgE staining, ELISPOT analysis provided evidence that the differentiation of IgE committed B cells to the plasma cell stage was arrested at high cell doses. In fact, splitting the cells at higher concentrations after culture initiation increased IgE production. Cells plated at higher doses were found to be more prone to apoptosis as assessed by Annexin staining. Interestingly, inhibiting apoptosis by the use of the caspase inhibitor DEVD significantly increased IgE levels implicating apoptosis in the preferential deletion of IgE expressing cells. These data not only highlight the caveat against using a single B-cell dose for IgE production in vitro but also suggest for the first time a possible IgE regulatory mechanism mediated by cell density.