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.
Cocca
Brian A
BA
Department of Molecular Sciences, University of Tennessee Health Sciences Center, Memphis, TN 38163, USA.
Cline
Amy M
AM
Radic
Marko Z
MZ
D017209:Apoptosis / Q000276:immunology*; D001324:Autoantigens / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology* / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism; D001666:Binding Sites, Antibody; D002462:Cell Membrane / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D002467:Cell Nucleus / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D053938:DNA Fragmentation / Q000276:immunology; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D019169:Jurkat Cells; D018613:Microscopy, Confocal; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D054460:rho-Associated Kinases
D001324:Autoantigens; D007135:Immunoglobulin Variable Region; D047908:Intracellular Signaling Peptides and Proteins; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases