African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12193

Full Length Research Paper

Regression of mouse-derived renal cancer by adoptive transfer of tumor-reactive RNAi-induced TGF-beta-insensitive CD8+ T cells

Bin Song1*, Wei-jun Qin2, Zeng-yue Yang1, Ting-yi Bao1, Xia-Yang3, Fu-li Wang2, Geng Zhang2, An-gang Yang4 and He Wang2
1Department of Urology, Tangdu Hospital, Fourth Military Medical University, Xin Si Road, Xi'an, 710038, China. 2Department of Urology, Xijing Hospital, Fourth Military Medical University, Chang Le West Road 15, Xi'an, 710032, China. 3Department of Division of Medical Service, Tangdu Hospital, Fourth Military Medical University, Xin Si Road, Xi'an, 710038, China. 4Department of Immunology, Fourth Military Medical University, Chang Le West Road 17, Xi'an, 710032, China.
Email: [email protected], [email protected]

  •  Published: 31 August 2010

Abstract

Transforming growth factor beta (TGF-beta) is a potent immunosuppressant. The present study was conducted to develop a treatment strategy through adoptive transfer of tumor-reactive RNAi-induced TGF-beta-insensitive CD8+ T cells. BALB/c mice were primed with irradiated Renca cells. CD8+ T cells were isolated from the spleen of primed animals, expanded ex vivo and were rendered TGF-beta-insensitive by infecting with a retrovirus containing shRNA to mouse TGF-beta type II receptor gene (MSCV-shRNA-T). Control CD8+ T cells consist of those infected with retroviruses containing shRNA to non specific gene (MSCV-shRNA-N) and naive CD8+ T cells. The effect of all groups of CD8+ T cells on Renca cells were analyzed by semi-quantitative RT-PCR, Western-blot, in vitro and in vivo assay. MSCV-shRNA-T group of CD8+ T cells were resistant to the antiproliferative effect of exogenous TGF-beta, while control groups were not. Results of Western blot showed the Smad pathway was disrupted in MSCV-shRNA-T group, which confirmed the blockade of the signal transduction pathway. In vitro cytotoxic assay revealed that these tumor-reactive, TGF-beta-insensitive CD8+ T cells killed Renca cells specifically and strongly. Adoptive transfer of these MSCV-shRNA-T CD8+ T cells to BALB/c tumor-bearing mice showed strong tumor-specific cytotoxic T lymphocyte responses and antitumor immunity against Renca renal cancer. Based on these results, we predict that adoptive transfer of tumor-reactive RNAi-induced TGF-beta-insensitive CD8+ T cells may be effective to renal cancer therapy.

 

Key words: Transforming growth factor beta, adoptive transfer, RNA interference, renal cancer, renca cells, immunotherapy.

Abbreviation

CM, Complete medium; CTL, cytotoxic thymus lymphocyte; FBS,fetal bovine serum; GFP, green fluorescent protein; TGF-ß, transforming growth factor beta; Renca, mouse-derived renal cancer; TßRI, type I TGF-ß receptor;TßRII, type II TGF-ß receptor; TßRIII, type III TGF-ß receptor; RNAi, RNA interference; GAPDH, glyceraldehyde-3-phosphate-dehydrogenase; MHC, major histocompatibility complex; MSCV, murine stem cell virus; DMEM, dulbecco's modification of eagle's medium; RT-PCR, reverse transcription-polymerase chain reaction.