van Nieuwenhuijze A E M, van Lopik T, Smeenk R J T, Aarden L A
Department of Immunopathology, Sanquin Research at CLB University of Amsterdam, 1006 AD Amsterdam, The Netherlands.
Ann Rheum Dis. 2003 Jan;62(1):10-4. doi: 10.1136/ard.62.1.10.
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.
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.
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.
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.
在体外系统中研究凋亡细胞的核小体泄漏动力学，并将结果外推至自身免疫性疾病，尤其是系统性红斑狼疮。
利用抗组蛋白3单克隆抗体（mAb）和抗核小体mAb开发了一种灵敏的核小体酶联免疫吸附测定（ELISA）。在Jurkat细胞中研究凋亡细胞死亡过程中的核小体释放。将细胞的膜联蛋白V结合（早期凋亡）和碘化丙啶阳性（晚期凋亡）与凋亡诱导后不同时间的核小体释放进行比较。
凋亡诱导后24至48小时，Jurkat细胞的培养上清液中出现核小体，此时细胞已处于晚期凋亡状态超过12小时。
核小体从晚期凋亡的Jurkat细胞中释放，比膜联蛋白V结合细胞出现延迟12小时。该结果表明体内清除机制有12小时从循环中清除凋亡物质。
