Cancer Immunology, Immunotherapy

, Volume 58, Issue 6, pp 977–987

Lentiviral vectors encoding human MUC1-specific, MHC-unrestricted single-chain TCR and a fusion suicide gene: potential for universal and safe cancer immunotherapy

  • Xiaochuan Chen
  • Wentao Gao
  • Andrea Gambotto
  • Olivera J. Finn
Original Article

DOI: 10.1007/s00262-008-0624-0

Cite this article as:
Chen, X., Gao, W., Gambotto, A. et al. Cancer Immunol Immunother (2009) 58: 977. doi:10.1007/s00262-008-0624-0

Abstract

MUC1 tumor antigen is a target for immunotherapy of most human adenocarcinomas and some hematological malignancies. Expression of a MUC1-specific, MHC-unrestricted single-chain T cell receptor (scTCR) on cells of both innate and adaptive immune system through reconstitution of lethally irradiated mice by retroviral vector-transduced bone marrow cells, had been shown to effectively control the growth of MUC1+ tumors independent of their MHC type, suggesting that this receptor is a good candidate for broadly applicable gene therapy/immunotherapy. However, the translational application of this immuno-gene therapy modality was discouraged by the progressive transgene silencing in reconstituted T and B cells, as well as the potential of tumorogenesis intrinsic to oncoretroviral vectors. To overcome these problems and facilitate the future clinical use of this receptor, we have constructed a panel of novel self-inactivating lentiviral vectors (LVs) which harbor two independent internal promoters, one driving expression of the scTCR gene and the other of a fusion suicide gene, the HSV-TK–EGFP fusion gene, allowing the transduced cells to be destroyable by the pro-drug ganciclovir. Despite the large size of insert, these vectors were efficiently packaged into high titer virus that transferred the expression of transgene in both T cell lines and primary T cells. Sustained expression was maintained in a T cell line for over 4 months in vitro, suggesting its efficient resistance to transgene silencing. Both scTCR and HSV-TK–EGFP genes were functional in the transduced cells, as evidenced by their specific recognition of MUC1+ tumors and efficient eradication by ganciclovir.

Keywords

MUC1T cell receptor (TCR)Lentiviral vectorCancer immunotherapy

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Xiaochuan Chen
    • 1
    • 2
  • Wentao Gao
    • 3
  • Andrea Gambotto
    • 4
  • Olivera J. Finn
    • 1
  1. 1.Department of ImmunologyUniversity of Pittsburgh School of MedicinePittsburghUSA
  2. 2.Garden State Cancer CenterCenter for Molecular Medicine and ImmunologyBellevilleUSA
  3. 3.Department of PathologyUniversity of Pittsburgh School of MedicinePittsburghUSA
  4. 4.Center for Biotechnology and Institute of Molecular MedicineUniversity of Pittsburgh School of MedicinePittsburghUSA