Homotypic interactions protect chronic lymphocytic leukaemia cells from spontaneous death in vitro

Chronic lymphocytic leukaemia (CLL) cells are long-lived in vivo but undergo spontaneous apoptosis when cultured in vitro. Since CLL cells associate intimately with one another at sites of tissue involvement, we examined the hitherto unproven possibility that homotypic interactions between the malignant cells might reduce their propensity to undergo spontaneous cell death. In a series of experiments in which highly pure CLL-cell populations were cultured on a non-adherent surface, cell viability was found to increase markedly with the level of crowding at the bottom of the culture vessel. The effect was observed among unevenly distributed cells within a single culture vessel and did not require direct cell-cell contact. This indicates that cell survival was being regulated in an autocrine fashion by locally acting soluble products. Conditioned medium from crowded CLL cells enhanced the survival of autologous non-crowded cells, indicating that at least some of the autocrine survival factors produced by CLL cells could accumulate in the extracellular environment. In addition, the survival of non-crowded CLL cells was markedly enhanced by co-culturing them with an excess of autologous fixed cells. This protective effect of direct cell-cell contact was mediated by specific surface structures since it was abrogated by pre-treating the fixed cells with neuraminidase. Our results provide the first direct demonstration that the survival of cultured CLL cells is enhanced by homotypic interactions. We speculate that these protective effects may contribute to the accumulation of CLL cells in vivo, and that further elucidation of the underlying mechanisms may lead to novel therapeutic strategies.