Biomedical subjects

Amy M Cline

Publications and source records attributed to Amy M Cline.

3 recordsLinked to original sources

PubMed2004

Apoptosis, subcellular particles, and autoimmunity.

Firm evidence links the process of apoptosis to the induction of autoimmune disease. However, questions remain regarding the precise interactions of dying cells with the immune system. Genetic analyses indicate that deficiencies in serum proteins or receptors that mediate clearance of apoptotic cells increase the risk of autoimmunity. Moreover, administration of apoptotic cells to naive animals elicits transient autoimmune responses. Because known autoantigens are covalently modified and redistributed to cell surface blebs during the execution stage of apoptosis, increasing attention is being directed at this stage of programmed cell death, and researchers have identified a variety of autoantigens that are sequestered within blebs. However, blebs are merely a transition stage toward the complete cellular fragmentation, as blebs quickly convert into apoptotic bodies, subcellular particles (SCPs) of heterogeneous size, surface composition, and cargo. Because certain types of subcellular particles represent packets of highly enriched autoantigens, we propose that they are relevant to our understanding of autoimmunity.

Animals↗

PubMed2004

Murine lupus autoantibodies identify distinct subsets of apoptotic bodies.

The specific modification of autoantigens and their redistribution into blebs at the surface of apoptotic cells contribute to the induction of autoimmune responses. Blebs containing fragments of the apoptotic nucleus separate from the remainder of the cell to form membrane-bound sub-cellular particles (SCPs), otherwise known as apoptotic bodies. To determine whether apoptotic bodies containing nuclear antigens represent a defined subset of SCPs, we examined the heterogeneity of particles generated by Jurkat cells following synchronization of the cell cycle by serum withdrawal and inhibition of topoisomerase I by camptothecin. Particles were purified by filtration, incubated in the presence of antinucleosome or anti-cardiolipin autoantibodies, annexin V, and Sytox Orange and analyzed by flow cytometry and confocal microscopy. We demonstrate that nuclear autoantigens are associated with one clearly defined subset of SCPs that can be distinguished from other products of late apoptosis. Our experiments represent an important step towards characterizing the heterogeneity of SCPs that are generated in late apoptosis and identifying their contributions to tolerance and autoimmunity.

Animals↗

PubMed2002

Blebs and apoptotic bodies are B cell autoantigens.

Mounting evidence suggests that systemic lupus erythematosus autoantigens are derived from apoptotic cells. To characterize the potential interactions between apoptotic cells and B cells, the D56R/S76R variant of 3H9, a murine autoantibody that binds to DNA, chromatin, and anionic phospholipids, was compared with DNA4/1, a human anti-DNA autoantibody. Flow cytometry revealed that only D56R/S76R bound to Jurkat cells treated with either of three distinct proapoptotic stimuli, Ab binding was dependent on caspase activity, and immunoreactivity developed subsequent to annexin V binding. Confocal microscopy established a structural basis for the distinct kinetics of binding. D56R/S76R preferentially bound to membrane blebs of apoptotic cells, whereas annexin V binding did not require blebs. Inhibition of ROCK I kinase, an enzyme that stimulates nuclear fragmentation and fragment distribution into blebs, significantly reduced Ab binding. Because members of the collectin and pentraxin families of serum proteins bind to blebs on apoptotic cells and assist in the clearance of cellular remains, our results suggest that Abs to blebs could affect the recognition of apoptotic cells by cells of the innate immune system and thus modify tolerance to nuclear Ags.

Apoptosis↗