OBJECTIVE: To investigate the kinetics of nucleosome leakage from apoptotic cells in an in vitro system and extrapolate the results to autoimmune disease, in particular systemic lupus erythematosus. METHODS: A sensitive nucleosome enzyme linked immunosorbent assay (ELISA) was developed, using a monoclonal antibody (mAb) against histone 3 and an mAb against nucleosomes. Nucleosome release during apoptotic cell death was studied in Jurkat cells. AnnexinV binding (early apoptosis) and propidium iodide positivity (late apoptosis) of the cells were compared with nucleosome release at different times after apoptosis induction. RESULTS: Nucleosomes appeared in culture supernatant of Jurkat cells 24 to 48 hours after apoptosis induction, when the cells had been late apoptotic for more than 12 hours. CONCLUSION: Nucleosomes are released from late apoptotic Jurkat cells, with a 12 hour delay from the appearance of AnnexinV binding cells. This result suggests that in vivo scavenger mechanisms have 12 hours to remove apoptotic material from the circulation.
OBJECTIVE: To investigate the kinetics of nucleosome leakage from apoptotic cells in an in vitro system and extrapolate the results to autoimmune disease, in particular systemic lupus erythematosus. METHODS: A sensitive nucleosome enzyme linked immunosorbent assay (ELISA) was developed, using a monoclonal antibody (mAb) against histone 3 and an mAb against nucleosomes. Nucleosome release during apoptotic cell death was studied in Jurkat cells. AnnexinV binding (early apoptosis) and propidium iodide positivity (late apoptosis) of the cells were compared with nucleosome release at different times after apoptosis induction. RESULTS: Nucleosomes appeared in culture supernatant of Jurkat cells 24 to 48 hours after apoptosis induction, when the cells had been late apoptotic for more than 12 hours. CONCLUSION: Nucleosomes are released from late apoptotic Jurkat cells, with a 12 hour delay from the appearance of AnnexinV binding cells. This result suggests that in vivo scavenger mechanisms have 12 hours to remove apoptotic material from the circulation.
Authors: Gerben Marsman; Helen von Richthofen; Ingrid Bulder; Florea Lupu; Jan Hazelzet; Brenda M Luken; Sacha Zeerleder Journal: Blood Adv Date: 2017-11-30
Authors: Roshanak Tolouei Semnani; Priyanka Goel Venugopal; Lily Mahapatra; Jason A Skinner; Francoise Meylan; Daniel Chien; David W Dorward; Damien Chaussabel; Richard M Siegel; Thomas B Nutman Journal: J Immunol Date: 2008-11-15 Impact factor: 5.422
Authors: Anne Jan van der Meer; Achmed Achouiti; Arie van der Ende; Aicha A Soussan; Sandrine Florquin; Alex de Vos; Sacha S Zeerleder; Tom van der Poll Journal: Mol Med Date: 2016-06-24 Impact factor: 6.354
Authors: Jill A Gallaher; Pedro M Enriquez-Navas; Kimberly A Luddy; Robert A Gatenby; Alexander R A Anderson Journal: Cancer Res Date: 2018-01-30 Impact factor: 12.701
Authors: F Stephan; I Dienava-Verdoold; I Bulder; D Wouters; A E Mast; H Te Velthuis; L A Aarden; S Zeerleder Journal: J Thromb Haemost Date: 2012-06 Impact factor: 5.824