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[Dna comets as markers of cells death]
V A Tronov1, T A Nikol'skaia, M A Konopliannikov
1Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow, Russia.
Biofizika
|July 27, 1999
Summary
DNA comets accurately detect apoptosis, a type of cell death. This method revealed higher apoptosis levels in systemic lupus erythematosus patients' lymphocytes, aiding in disease research.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Context:
- Human peripheral blood lymphocytes undergo spontaneous apoptosis in vitro.
- Morphological assessment confirmed apoptosis as the primary mode of cell death.
- Standard cell death assays can be complemented by novel molecular techniques.
Purpose:
- To evaluate DNA comets as reliable biomarkers for detecting apoptosis in lymphocytes.
- To correlate DNA comet characteristics with morphological signs of apoptosis.
- To investigate apoptosis levels in lymphocytes from patients with systemic lupus erythematosus (SLE).
Summary:
- Lymphocytes undergoing apoptosis in vitro were analyzed using the single-cell gel electrophoresis (SCGE) assay to form DNA comets.
- DNA comet parameters (DNA content, tail length, tail moment, DNA fraction in tail) were used to classify comets.
- A strong correlation was observed between the fraction of morphologically apoptotic cells and the fraction of "apoptotic" DNA comets, validating the assay's efficacy.
- The study identified an elevated level of in vitro apoptosis in lymphocytes from SLE patients using DNA comets as markers.
Impact:
- Establishes DNA comet assay as a sensitive and quantitative method for assessing lymphocyte apoptosis.
- Provides a tool for further research into the mechanisms of spontaneous cell death.
- Highlights potential utility in studying immune dysregulation in autoimmune diseases like SLE.