Methods for inducing apoptosis.
Apoptotic cells are sources of tolerogenic material during tissue homeostasis; abnormalities in apoptosis or in the clearance of apoptotic material generate a novel source of antigens against which an autoimmune response may be initiated. In our laboratory, we study the biochemistry and cell biology of systemic autoimmune disease autoantigens during different forms of cell death. Several different methods for inducing apoptosis, and for assaying the induction of this cellular process, are routinely performed. This chapter describes methods for inducing apoptosis via ultraviolet B irradiation, small molecule drug treatments, death receptor ligation, and exposure to granule components of cytotoxic lymphocytes. Assays to confirm the induction of apoptosis by quantifying changes in mitochondrial membrane potential, phosphatidylserine membrane localization, DNA content, and autoantigen cleavage are also detailed.
Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Kathryn M
KM
Roberts
Antony
A
Rosen
Livia A
LA
Casciola-Rosen
D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology* / Q000528:radiation effects; D001324:Autoantigens / Q000378:metabolism; D015551:Autoimmunity; D002453:Cell Cycle; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D015979:Killer Cells, Lymphokine-Activated / Q000276:immunology; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D013194:Staining and Labeling; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014466:Ultraviolet Rays; D019014:fas Receptor / Q000378:metabolism
D017304:Annexin A5; D001324:Autoantigens; D019014:fas Receptor