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Related Experiment Videos

Lymphocyte apoptosis: induction by gene transfer techniques

O Ebert1, S Finke, A Salahi

  • 1Abteilung für Innere Medizin, Humboldt-Universität zu Berlin.

Gene Therapy
|April 1, 1997
PubMed
Summary
This summary is machine-generated.

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Gene transfer into lymphocytes is difficult due to cell death. Blocking tumor necrosis factor (TNF) reduces apoptosis and necrosis, improving gene transfer efficiency for cancer immunotherapy.

Area of Science:

  • Immunology
  • Molecular Biology
  • Gene Therapy

Background:

  • Efficient gene transfer into lymphocytes is a significant challenge.
  • The molecular mechanisms underlying lymphocyte resistance to gene transfer are not fully understood.
  • Apoptosis (programmed cell death) is hypothesized to contribute to this resistance.

Purpose of the Study:

  • To investigate the role of apoptosis in lymphocyte gene transfer resistance.
  • To examine the correlation between gene transfer methods, tumor necrosis factor (TNF) secretion, and lymphocyte apoptosis.
  • To evaluate the potential of blocking TNF to improve gene transfer outcomes.

Main Methods:

  • Lymphocytes were transfected using various nonviral and viral gene transfer methods.
  • Apoptosis, necrosis, and TNF-alpha secretion levels were measured in transfected lymphocytes.

Related Experiment Videos

  • The effect of anti-TNF antibodies on apoptosis and lymphocyte proliferation was assessed.

    • Main Results:

    • Nonviral gene transfer methods, particularly receptor-mediated gene transfer and lipofection, induced significant apoptosis and TNF-alpha secretion in lymphocytes.
    • Electroporation and retroviral gene transfer resulted in lower TNF-alpha secretion.
    • Blocking TNF-alpha with an antibody significantly reduced apoptosis and necrosis, leading to increased proliferation of lymphocytes engineered with cytokine genes (interleukin-2 or interleukin-7).

    Conclusions:

    • Gene transfer into lymphocytes triggers TNF-alpha secretion, leading to apoptosis and necrosis.
    • Targeting the TNF-alpha pathway can mitigate cell death during lymphocyte gene transfer.
    • Blocking apoptosis enhances the viability and proliferation of gene-modified lymphocytes, with implications for their use in cancer gene therapy.