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November 1, 2001The Journal of ImmunologyOpen Access

Caspase 8 Activity in Membrane Blebs After Anti-Fas Ligation

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Authors

BPBeverly Z. PackardOncoImmune (United States)AKAkira KomoriyaOncoImmune (United States)TBTilmann M. BrotzGynuity Health Projects

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Implication

In vitro imaging demonstrates early caspase 8 localization in plasma membrane blebs during Fas-mediated apoptosis in T cells, indicating localized initiation of proteolysis.

Key Points

  • To investigate the timing and spatial distribution of caspase 8 activation during anti-Fas-mediated apoptosis in cytotoxic T lymphocytes.
  • Induced apoptosis in cytotoxic T lymphocytes using anti-Fas ligation or interleukin-2 withdrawal.
  • Tracked dynamic caspase activation using cell-permeable fluorogenic peptide substrates selective for caspases 8 (IETDase), 9 (LEHDase), 1 (YVHDase), and 6 (VEIDase) via flow cytometry and confocal microscopy.
  • Flow cytometry initially indicated delayed caspase 8 (IETDase) activation relative to caspases 9, 1, and 6 in anti-Fas-treated cytotoxic T lymphocytes.
  • Confocal microscopy revealed that caspase 8 was actually activated early but segregated into shed plasma membrane vesicles that were excluded from conventional flow cytometry measurements.
  • Apoptosis induced via interleukin-2 withdrawal showed early cytoplasmic caspase 8 activity without the formation of plasma membrane vesicles.

Cite This Study

Packard et al. (2001) studied this question.

synapsesocial.com/papers/6aa8f9957ea61a08df43ec49https://doi.org/10.4049/jimmunol.167.9.5061
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Also Consider

Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context:

  1. 1Assessment of Caspase Activities in Intact Apoptotic Thymocytes Using Cell-Permeable Fluorogenic Caspase Substrates2000 · 165 citations
  2. 2MATURE T LYMPHOCYTE APOPTOSIS—Immune Regulation in a Dynamic and Unpredictable Antigenic Environment1999 · 950 citations
  3. 3Caspase-3 controls both cytoplasmic and nuclear events associated with Fas-mediated apoptosis<i>in vivo</i>1998 · 244 citations
  4. 4Proteolysis of Fodrin (Non-erythroid Spectrin) during Apoptosis1995 · 533 citations
  5. 5Apoptotic Membrane Blebbing Is Regulated by Myosin Light Chain Phosphorylation1998 · 496 citations