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Publication year
2011Source
Apoptosis, 16, 1, (2011), pp. 86-93ISSN
Publication type
Article / Letter to editor
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Organization
Nephrology
Journal title
Apoptosis
Volume
vol. 16
Issue
iss. 1
Page start
p. 86
Page end
p. 93
Subject
NCMLS 1: Infection and autoimmunity N4i 4: Auto-immunity, transplantation and immunotherapy; NCMLS 2: Immune Regulation N4i 4: Auto-immunity, transplantation and immunotherapy; NCMLS 3: Tissue engineering and pathology N4i 4: Auto-immunity, transplantation and immunotherapyAbstract
In our research on the role of apoptosis in the pathogenesis of the autoimmune disease systemic lupus erythematosus (SLE), we aim to evaluate the effects of early and late apoptotic cells and blebs on antigen presenting cells. This requires the in vitro generation of sufficiently large and homogeneous populations of early and late apoptotic cells. Here, we present a quick method encountered by serendipity that results in highly reproducible synchronized homogeneous apoptotic cell populations. In brief, granulocytic 32Dcl3 cells are incubated on ice for 2 h and subsequently rewarmed at 37 degrees C. After 30-90 min at 37 degrees C more than 80-90% of the cells become early apoptotic (Annexin V positive/propidium iodide negative). After 24 h of rewarming at 37 degrees C 98% of the cells were late apoptotic (secondary necrotic; Annexin V positive/propidium iodide positive). Cells already formed apoptotic blebs at their cell surface after approximately 20 min at 37 degrees C. Inter-nucleosomal chromatin cleavage and caspase activation were other characteristics of this cold-shock-induced process of apoptosis. Consequently, apoptosis could be inhibited by a caspase inhibitor. Finally, SLE-derived anti-chromatin autoantibodies showed a high affinity for apoptotic blebs generated by cold-shock. Overall, cold-shock induced apoptosis is achieved without the addition of toxic compounds or antibodies, and quickly leads to synchronized homogeneous apoptotic cell populations, which can be applied for various research questions addressing apoptosis.
This item appears in the following Collection(s)
- Academic publications [238441]
- Electronic publications [122508]
- Faculty of Medical Sciences [90373]
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