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Induction of apoptosis in experimental human B cell lymphomas by conditional TRAIL-expressing T cells.

Ucur E, Mattern J, Wenger T, Okouoyo S, Schroth A, Debatin KM, Herr I - Br. J. Cancer (2003)

Bottom Line: Jurkat-TRAIL cells had an additive effect with cytotoxic drugs in vitro, since cell death was enhanced.To elucidate the antitumoral activity of these Jurkat-TRAIL cells in vivo, we injected them intratumorally in xenografts of human Burkitt lymphomas.Switching on expression of TRAIL by adding tetracycline to the drinking water of the mice strongly reduced tumour growth by apoptosis in a caspase-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Clinical Cooperation Unit, Molecular Oncology/Pediatrics, German Cancer Research Center, Heidelberg, Germany.

ABSTRACT
In the present study, we demonstrate the utility of a non-tumour-forming T-cell line for the inducible gene transfer of tumour necrosis factor (TNF)-related apoptosis-inducing ligand (Apo2L/TRAIL), which has been shown to selectively induce apoptosis in malignant but not in normal cells. To generate T cells inducible for TRAIL expression, we stably transfected Jurkat cells with TRAIL in the context of the Tet-On system. The switched on cells strongly expressed TRAIL mRNA, whose protein product was expressed on the cell surface. Paracrine induction of apoptosis in human target tumour cells was solely found for membrane-bound TRAIL. The Jurkat-TRAIL cells itself survived due to clonal selection of TRAIL-resistant cells. Jurkat-TRAIL cells had an additive effect with cytotoxic drugs in vitro, since cell death was enhanced. To elucidate the antitumoral activity of these Jurkat-TRAIL cells in vivo, we injected them intratumorally in xenografts of human Burkitt lymphomas. Switching on expression of TRAIL by adding tetracycline to the drinking water of the mice strongly reduced tumour growth by apoptosis in a caspase-dependent manner. Thus, non-tumour-forming T-cell lines offer a novel method for gene transfer and inducible expression of TRAIL in tumour therapy.

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Jurkat-TRAIL cells inhibit tumour growth of xenografted BJAB cells. (A) Schematic representation of the experiment. Athymic nude mice were xenografted with 5 × 107 BJAB cells. After 3 days, 5 × 107 Jurkat-TRAIL or Jurkat-CO cells were injected intratumorally. To switch on the Tet-system, tet (1 mg ml−1) was added to the drinking water, which was sweatened with 5% glucose for the duration of the experiment. (B) Upper panel: tumour growth of BJAB xenografts, injected with switched on Jurkat-TRAIL (black bars) or Jurkat-CO cells, was weekly measured during a period of 4 weeks. Data are presented as the means of eight animals and s.d. are shown (*=13.08). Lower panel: nude mice were xenografted with 5 × 107 Kelly cells and treated as described above. Data are presented as the means of four animals and s.d. is shown. (C) Cryosections of BJAB xenografts harvested 3 days upon switching on TRAIL expression were stained with a specific rabbit polyclonal anti-TRAIL antibody, followed by immunofluorescence detection. Staining or the absence of the primary antibody served as control (CO). The representative results shown are from one of three different experiments.
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fig5: Jurkat-TRAIL cells inhibit tumour growth of xenografted BJAB cells. (A) Schematic representation of the experiment. Athymic nude mice were xenografted with 5 × 107 BJAB cells. After 3 days, 5 × 107 Jurkat-TRAIL or Jurkat-CO cells were injected intratumorally. To switch on the Tet-system, tet (1 mg ml−1) was added to the drinking water, which was sweatened with 5% glucose for the duration of the experiment. (B) Upper panel: tumour growth of BJAB xenografts, injected with switched on Jurkat-TRAIL (black bars) or Jurkat-CO cells, was weekly measured during a period of 4 weeks. Data are presented as the means of eight animals and s.d. are shown (*=13.08). Lower panel: nude mice were xenografted with 5 × 107 Kelly cells and treated as described above. Data are presented as the means of four animals and s.d. is shown. (C) Cryosections of BJAB xenografts harvested 3 days upon switching on TRAIL expression were stained with a specific rabbit polyclonal anti-TRAIL antibody, followed by immunofluorescence detection. Staining or the absence of the primary antibody served as control (CO). The representative results shown are from one of three different experiments.

Mentions: Since Jurkat cells itself do not expand and do not form tumours in mice (unpublished observation), we assessed the effect of Jurkat-TRAIL cells in an in vivo tumour model. Athymic nude mice were xenografted with BJAB cells. After 3 days, at tumour volumes of about 100 mm3, animals were injected intratumorally with switched off Jurkat-TRAIL or Jurkat-CO cells. Tet was added to the drinking water of the mice to switch on the system (Figure 5AFigure 5


Induction of apoptosis in experimental human B cell lymphomas by conditional TRAIL-expressing T cells.

Ucur E, Mattern J, Wenger T, Okouoyo S, Schroth A, Debatin KM, Herr I - Br. J. Cancer (2003)

Jurkat-TRAIL cells inhibit tumour growth of xenografted BJAB cells. (A) Schematic representation of the experiment. Athymic nude mice were xenografted with 5 × 107 BJAB cells. After 3 days, 5 × 107 Jurkat-TRAIL or Jurkat-CO cells were injected intratumorally. To switch on the Tet-system, tet (1 mg ml−1) was added to the drinking water, which was sweatened with 5% glucose for the duration of the experiment. (B) Upper panel: tumour growth of BJAB xenografts, injected with switched on Jurkat-TRAIL (black bars) or Jurkat-CO cells, was weekly measured during a period of 4 weeks. Data are presented as the means of eight animals and s.d. are shown (*=13.08). Lower panel: nude mice were xenografted with 5 × 107 Kelly cells and treated as described above. Data are presented as the means of four animals and s.d. is shown. (C) Cryosections of BJAB xenografts harvested 3 days upon switching on TRAIL expression were stained with a specific rabbit polyclonal anti-TRAIL antibody, followed by immunofluorescence detection. Staining or the absence of the primary antibody served as control (CO). The representative results shown are from one of three different experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376862&req=5

fig5: Jurkat-TRAIL cells inhibit tumour growth of xenografted BJAB cells. (A) Schematic representation of the experiment. Athymic nude mice were xenografted with 5 × 107 BJAB cells. After 3 days, 5 × 107 Jurkat-TRAIL or Jurkat-CO cells were injected intratumorally. To switch on the Tet-system, tet (1 mg ml−1) was added to the drinking water, which was sweatened with 5% glucose for the duration of the experiment. (B) Upper panel: tumour growth of BJAB xenografts, injected with switched on Jurkat-TRAIL (black bars) or Jurkat-CO cells, was weekly measured during a period of 4 weeks. Data are presented as the means of eight animals and s.d. are shown (*=13.08). Lower panel: nude mice were xenografted with 5 × 107 Kelly cells and treated as described above. Data are presented as the means of four animals and s.d. is shown. (C) Cryosections of BJAB xenografts harvested 3 days upon switching on TRAIL expression were stained with a specific rabbit polyclonal anti-TRAIL antibody, followed by immunofluorescence detection. Staining or the absence of the primary antibody served as control (CO). The representative results shown are from one of three different experiments.
Mentions: Since Jurkat cells itself do not expand and do not form tumours in mice (unpublished observation), we assessed the effect of Jurkat-TRAIL cells in an in vivo tumour model. Athymic nude mice were xenografted with BJAB cells. After 3 days, at tumour volumes of about 100 mm3, animals were injected intratumorally with switched off Jurkat-TRAIL or Jurkat-CO cells. Tet was added to the drinking water of the mice to switch on the system (Figure 5AFigure 5

Bottom Line: Jurkat-TRAIL cells had an additive effect with cytotoxic drugs in vitro, since cell death was enhanced.To elucidate the antitumoral activity of these Jurkat-TRAIL cells in vivo, we injected them intratumorally in xenografts of human Burkitt lymphomas.Switching on expression of TRAIL by adding tetracycline to the drinking water of the mice strongly reduced tumour growth by apoptosis in a caspase-dependent manner.

View Article: PubMed Central - PubMed

Affiliation: Clinical Cooperation Unit, Molecular Oncology/Pediatrics, German Cancer Research Center, Heidelberg, Germany.

ABSTRACT
In the present study, we demonstrate the utility of a non-tumour-forming T-cell line for the inducible gene transfer of tumour necrosis factor (TNF)-related apoptosis-inducing ligand (Apo2L/TRAIL), which has been shown to selectively induce apoptosis in malignant but not in normal cells. To generate T cells inducible for TRAIL expression, we stably transfected Jurkat cells with TRAIL in the context of the Tet-On system. The switched on cells strongly expressed TRAIL mRNA, whose protein product was expressed on the cell surface. Paracrine induction of apoptosis in human target tumour cells was solely found for membrane-bound TRAIL. The Jurkat-TRAIL cells itself survived due to clonal selection of TRAIL-resistant cells. Jurkat-TRAIL cells had an additive effect with cytotoxic drugs in vitro, since cell death was enhanced. To elucidate the antitumoral activity of these Jurkat-TRAIL cells in vivo, we injected them intratumorally in xenografts of human Burkitt lymphomas. Switching on expression of TRAIL by adding tetracycline to the drinking water of the mice strongly reduced tumour growth by apoptosis in a caspase-dependent manner. Thus, non-tumour-forming T-cell lines offer a novel method for gene transfer and inducible expression of TRAIL in tumour therapy.

Show MeSH
Related in: MedlinePlus