Serum-dependent Processing of Late Apoptotic Cells and Their Immunogenicity

The execution phase of apoptosis involves many processes which modify cellular molecules for an efficient and quiet elimination of the dead cell. These include exposure and secretion of "eat-me" signals, to attract phagocytes, as well as degradation of immune-stimulating cell debris. During this phase apoptotic microparticles (MPs) are released from the dying cell. The remaining cell remnant forms large late apoptotic cell-derived membranous vesicles (ACMV(L)) on its surface which remain attached. Phagocytosis is enhanced by cell non-autonomous factors such as complement component C1q and serum DNase I. We studied the formation and retraction of ACMV(L) and the influence of serum on their dynamics. We furthermore investigated the immunogenicity of cell remnants compared to released MPs. ACMV(L) were examined using time-lapse, electron microscopy and confocal microscopy. These blebs were observed on cell remnants with intact and with permeable membrane. This suggests that ACMV(L) remain on the surface by the time the cell remnant enters secondary necrosis. Bleb retraction could also be observed, but was radically enhanced in the presence of serum. Additionally, MPs stimulate peripheral blood mononuclear cells to produce similar IL-1beta, IL-6, IL-8, IL-10, and TNF-alpha levels as LPS. In contrast, cell remnants only induce high levels of IL-8. These data show that cell non-autonomous factors contribute to morphological rearrangements during late apoptosis. In addition, they implicate that apoptotic MPs are released to attract phagocytes, while apoptotic cell remnants further process their potentially immunogenic content to prevent an inflammatory response upon secondary necrosis.