Suppression of apoptosis in a cytotoxic T-cell line by interleukin 2-mediated gene transcription and deregulated expression of the protooncogene bcl-2

Ge Deng, Eckhard R. Podack

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167 Citations (Scopus)

Abstract

Absence of interleukin 2 (IL-2) from IL-2-dependent cells, such as the cytotoxic T-cell line CTLL2, causes DNA fragmentation and programmed cell death (apoptosis). We found that, upon initiation, DNA degradation proceeds rapidly. IL-2-deprived CTLL2 cells can be rescued from apoptosis by the addition of IL-2 2 h prior to the onset of detectable DNA breakdown. Addition of inhibitors of transcription with IL-2 abolished the IL-2-mediated rescue of CTLL2 cells. Thus it appears that IL-2-mediated gene transcription is necessary for survival. Deregulated expression of a protooncogene, bcl-2, inhibits apoptosis of cells dependent on other hematopoietic growth factors. To determine whether bcl-2 was active in CTLL2 cells, we transfected CTLL2 cells with a plasmid containing bcl-2 cDNA expressed under the metallothionein promoter and observed prolonged survival of the transfected cells upon IL-2 deprivation. Cell growth, however, was arrested in the G0/G1 or G2/M phases of the cell cycle. The prolonged survival of bcl-2 transfectants allowed the analysis of endogenous bcl-2 mRNA levels by Northern blot analysis. The expression of endogenous bcl-2 was down-regulated within 8 h of IL-2 withdrawal and was not detected after 3 days. Addition of IL-2 induced endogenous bcl-2 expression within 8 h. Full recovery of bcl-2 expression was achieved by 24 h after IL-2 addition. We conclude that the survival of death-prone CTLL2 cells may be viewed as IL-2-dependent suppression of suicide, probably by the IL-2-induced expression of the cellular bcl-2 gene.

Original languageEnglish
Pages (from-to)2189-2193
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number6
StatePublished - Mar 15 1993
Externally publishedYes

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Interleukin-2
Apoptosis
T-Lymphocytes
Cell Line
Genes
bcl-2 Genes
Metallothionein
G2 Phase
DNA
DNA Fragmentation
Northern Blotting
Cell Division
Suicide
Cell Survival
Intercellular Signaling Peptides and Proteins
Cell Cycle
Plasmids
Cell Death
Complementary DNA
Messenger RNA

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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abstract = "Absence of interleukin 2 (IL-2) from IL-2-dependent cells, such as the cytotoxic T-cell line CTLL2, causes DNA fragmentation and programmed cell death (apoptosis). We found that, upon initiation, DNA degradation proceeds rapidly. IL-2-deprived CTLL2 cells can be rescued from apoptosis by the addition of IL-2 2 h prior to the onset of detectable DNA breakdown. Addition of inhibitors of transcription with IL-2 abolished the IL-2-mediated rescue of CTLL2 cells. Thus it appears that IL-2-mediated gene transcription is necessary for survival. Deregulated expression of a protooncogene, bcl-2, inhibits apoptosis of cells dependent on other hematopoietic growth factors. To determine whether bcl-2 was active in CTLL2 cells, we transfected CTLL2 cells with a plasmid containing bcl-2 cDNA expressed under the metallothionein promoter and observed prolonged survival of the transfected cells upon IL-2 deprivation. Cell growth, however, was arrested in the G0/G1 or G2/M phases of the cell cycle. The prolonged survival of bcl-2 transfectants allowed the analysis of endogenous bcl-2 mRNA levels by Northern blot analysis. The expression of endogenous bcl-2 was down-regulated within 8 h of IL-2 withdrawal and was not detected after 3 days. Addition of IL-2 induced endogenous bcl-2 expression within 8 h. Full recovery of bcl-2 expression was achieved by 24 h after IL-2 addition. We conclude that the survival of death-prone CTLL2 cells may be viewed as IL-2-dependent suppression of suicide, probably by the IL-2-induced expression of the cellular bcl-2 gene.",
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T1 - Suppression of apoptosis in a cytotoxic T-cell line by interleukin 2-mediated gene transcription and deregulated expression of the protooncogene bcl-2

AU - Deng, Ge

AU - Podack, Eckhard R.

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N2 - Absence of interleukin 2 (IL-2) from IL-2-dependent cells, such as the cytotoxic T-cell line CTLL2, causes DNA fragmentation and programmed cell death (apoptosis). We found that, upon initiation, DNA degradation proceeds rapidly. IL-2-deprived CTLL2 cells can be rescued from apoptosis by the addition of IL-2 2 h prior to the onset of detectable DNA breakdown. Addition of inhibitors of transcription with IL-2 abolished the IL-2-mediated rescue of CTLL2 cells. Thus it appears that IL-2-mediated gene transcription is necessary for survival. Deregulated expression of a protooncogene, bcl-2, inhibits apoptosis of cells dependent on other hematopoietic growth factors. To determine whether bcl-2 was active in CTLL2 cells, we transfected CTLL2 cells with a plasmid containing bcl-2 cDNA expressed under the metallothionein promoter and observed prolonged survival of the transfected cells upon IL-2 deprivation. Cell growth, however, was arrested in the G0/G1 or G2/M phases of the cell cycle. The prolonged survival of bcl-2 transfectants allowed the analysis of endogenous bcl-2 mRNA levels by Northern blot analysis. The expression of endogenous bcl-2 was down-regulated within 8 h of IL-2 withdrawal and was not detected after 3 days. Addition of IL-2 induced endogenous bcl-2 expression within 8 h. Full recovery of bcl-2 expression was achieved by 24 h after IL-2 addition. We conclude that the survival of death-prone CTLL2 cells may be viewed as IL-2-dependent suppression of suicide, probably by the IL-2-induced expression of the cellular bcl-2 gene.

AB - Absence of interleukin 2 (IL-2) from IL-2-dependent cells, such as the cytotoxic T-cell line CTLL2, causes DNA fragmentation and programmed cell death (apoptosis). We found that, upon initiation, DNA degradation proceeds rapidly. IL-2-deprived CTLL2 cells can be rescued from apoptosis by the addition of IL-2 2 h prior to the onset of detectable DNA breakdown. Addition of inhibitors of transcription with IL-2 abolished the IL-2-mediated rescue of CTLL2 cells. Thus it appears that IL-2-mediated gene transcription is necessary for survival. Deregulated expression of a protooncogene, bcl-2, inhibits apoptosis of cells dependent on other hematopoietic growth factors. To determine whether bcl-2 was active in CTLL2 cells, we transfected CTLL2 cells with a plasmid containing bcl-2 cDNA expressed under the metallothionein promoter and observed prolonged survival of the transfected cells upon IL-2 deprivation. Cell growth, however, was arrested in the G0/G1 or G2/M phases of the cell cycle. The prolonged survival of bcl-2 transfectants allowed the analysis of endogenous bcl-2 mRNA levels by Northern blot analysis. The expression of endogenous bcl-2 was down-regulated within 8 h of IL-2 withdrawal and was not detected after 3 days. Addition of IL-2 induced endogenous bcl-2 expression within 8 h. Full recovery of bcl-2 expression was achieved by 24 h after IL-2 addition. We conclude that the survival of death-prone CTLL2 cells may be viewed as IL-2-dependent suppression of suicide, probably by the IL-2-induced expression of the cellular bcl-2 gene.

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