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TNF-alpha-induced apoptosis in Daudi cells: multiparametric analysis
R Di Pietro1, E Santavenere, L Centurione
1Istituto di Morfologia Umana Normale, Chieti, Italy.
Cytokine
|July 1, 1997
Summary
Tumour necrosis factor alpha (TNF-alpha) induces apoptosis in Daudi lymphoma cells within 6 hours. This programmed cell death occurs via alternative pathways, distinct from typical DNA fragmentation.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- Tumour necrosis factor alpha (TNF-alpha) is a key cytokine modulating immune responses.
- Understanding TNF-alpha's role in cell death is crucial for immunology and cancer research.
Purpose of the Study:
- To investigate the mechanism of TNF-alpha-induced apoptosis in human lymphoma (Daudi) cells.
- To characterize the morphological and molecular changes associated with TNF-alpha treatment over time.
Main Methods:
- Incubation of Daudi cells with TNF-alpha for 1, 6, and 24 hours.
- Analysis using vital DNA stains, DNA gel electrophoresis, in situ nick translation, transmission electron microscopy, and flow cytometry.
Main Results:
- Apoptosis was observed as early as 6 hours post-treatment, characterized by cell shrinkage and chromatin condensation.
- Apoptotic bodies formed without typical low molecular weight DNA fragmentation into nucleosomes.
- Nick translation assay showed 60% positive cells at 6 hours, decreasing to 40% at 24 hours.
- Flow cytometry did not reveal a subdiploid peak, indicating no classical DNA degradation.
Conclusions:
- TNF-alpha induces apoptosis in Daudi cells through alternative pathways.
- DNA degradation in TNF-alpha-induced apoptosis may not always involve oligonucleosomal fragmentation.