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Engineered CD8+ cytotoxic T cells with fiber-modified adenovirus-mediated TNF-α gene transfection counteract immunosuppressive interleukin-10-secreting lung metastasis and solid tumors

Cancer Gene Therapy volume 14, pages 661–675 (2007)Cite this article

Abstract

T-cell suppression derived from tumor-secreted immunosuppressive interleukin (IL)-10 becomes a major barrier to CD8+ T-cell immunotherapy of tumors. Tumor necrosis factor-alpha (TNF-α) is a multifunctional cytokine capable of activating T and dendritic cells (DCs) and counteracting IL-10-mediated DC inhibition and regulatory T-cell-mediated immune suppression. In this study, we constructed a recombinant adenovirus MFAdVTNF with fiber-gene modified by RGD insertion into the viral knob's H1 loop and a melanoma cell line B16OVA/IL−10 engineered to express ovalbumin (OVA) and to secrete IL-10 (2.2 ng/ml/106 cells/24 h). We transfected OVA-specific CD8+ T cells with MFAdVTNF, and found a fivefold increase in transgene human TNF-α secretion (4.3 ng/ml/106 cells/24 h) by the engineered CD8+ TTNF cells transfected with MFAdVTNF, compared to that (0.8 ng/ml/106 cells/24 h) by CD8+ T cells transfected with the original AdVTNF without viral fiber modification. The engineered CD8+ TTNF cells exhibited enhanced cytotoxicity and elongated survival in vivo after adoptive transfer. TNF-α derived from both the donor CD8+ T cells and the host cells plays an important role in donor CD8+ T-cell survival in vivo after adoptive transfer. We also demonstrated that the transfected B16OVA/IL−10 tumor cells secreting IL-10 are more resistant to in vivo CD8+ T-cell therapy than the original B16OVA tumor cells without IL-10 expression. Interestingly, the engineered CD8+ TTNF cells secreting transgene-coded TNF-α, but not the control CD8+ Tcontrol cells without any transgene expression eradicated IL-10-secreting 12-day lung micrometastasis in all 10/10 mice and IL-10-secreting solid tumors (∼5 mm in diameter) in 6/10 mice. Transfer of the engineered CD8+ TTNF cells further induced both donor- and host-derived memory CD8+ T cells, leading to a stronger long-term antitumor immunity against the IL-10-secreting B16OVA/IL−10 tumor cell challenges. Therefore, CD8+ T cells engineered to secrete TNF-α may be useful when designing strategies for adoptive T-cell therapy of solid tumors.

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Acknowledgements

This study was supported by research grants (MOP 67230/81228) of the Canadian Institutes of Health Research to JX. We thank Mark Boyd for his excellent technical support of flow cytometric analysis.

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Authors and Affiliations

  1. Health Research Division, Research Unit, Saskatchewan Cancer Agency, Saskatoon, Saskatchewan, Canada

    Z Ye, M Shi, T Chan, S Sas, S Xu & J Xiang

  2. Department of Oncology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada

    Z Ye, M Shi, T Chan, S Sas, S Xu & J Xiang

  3. Department of Immunology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada

    Z Ye, M Shi, T Chan, S Sas, S Xu & J Xiang

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Ye, Z., Shi, M., Chan, T. et al. Engineered CD8+ cytotoxic T cells with fiber-modified adenovirus-mediated TNF-α gene transfection counteract immunosuppressive interleukin-10-secreting lung metastasis and solid tumors. Cancer Gene Ther 14, 661–675 (2007). https://doi.org/10.1038/sj.cgt.7701039

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