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Homing and survival of thymidine kinase-transduced human T cells in NOD/SCID mice

Cancer Gene Therapy volume 9, pages 756–761 (2002)Cite this article

Abstract

The herpes simplex virus thymidine kinase (HSV-tk) gene conferring ganciclovir (GCV)-specific sensitivity to transduced cells might control Graft-versus-Leukemia (GvL)/Graft-versus-Host Disease (GvHD). Human T lymphocytes were engineered with an LSN-tk retroviral vector encoding tk and neomycin resistance (NeoR) genes. A total of 80×106 tk+ lymphocytes were injected intraperitoneally in NOD-SCID mice. Engraftment was evaluated by human CD45+/CD3+ cytofluorimetric analysis and NeoR-based polymerase chain reaction (PCR) on peripheral blood, bone marrow, liver, thymus, and spleen on day +5. After 14 days, GCV (10 mg/kg daily) cytofluorimetric analysis and PCR were repeated (day +19). Immunohistological studies with anti-CD3 monoclonal antibody followed by alkaline phosphatase and monoclonal anti–alkaline phosphatase staining were performed on spleen and liver at the same time points. Human CD45+/CD3+ cells were engrafted in all tissues on day +5 according to cytofluorimetry, immunohistology, and PCR. Lymphocytes “homed” to the white pulp T-cell area and to the red pulp; liver localization is prevalently at the periportal area. After GCV (day +19), cytofluorimetry and immunohistology showed very few CD3+ cells. PCR identified the transgene in 22% tissue samples (positive only in thymus and spleen). GvHD did not occur in any animal. These data demonstrate elevated doses of human-transduced CD3+ cells engraft in NOD/SCID mice; after GCV, very few CD3+ cells can be detected and those that escape treatment can be found in the thymus and in the spleen on day +19. Lack of full response to GCV may account for cases of GvHD in patients receiving tk-transduced T lymphocytes.

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Acknowledgements

We thank Geraldine Anne Boyd for her helpful criticism and comments. This research was supported by the Italian Association of Cancer Research (AIRC).

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Authors and Affiliations

  1. Department of Clinical and Experimental Medicine, Hematology and Clinical Immunology Section, Perugia University, Perugia, Italy

    Mauro Di Ianni, Adelmo Terenzi, Franca Falzetti, Sabrina Di Florio, Roberta Benedetti, Gigliola Venditti, Diego Alfonsi, Mariangela De Ioanni, Brunangelo Falini & Antonio Tabilio

  2. Department of Clinical and Experimental Medicine, Pathology Section, Perugia University, Perugia, Italy

    Andrea Bartoli

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  1. Mauro Di Ianni
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Correspondence to Antonio Tabilio.

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Di Ianni, M., Terenzi, A., Falzetti, F. et al. Homing and survival of thymidine kinase-transduced human T cells in NOD/SCID mice. Cancer Gene Ther 9, 756–761 (2002). https://doi.org/10.1038/sj.cgt.7700495

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