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

Cancer Gene Ther. 2002 Sep;9(9):756-61. doi: 10.1038/sj.cgt.7700495.


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 x 10(6) 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / metabolism
  • Antiviral Agents / pharmacology
  • Bone Marrow / immunology
  • Cell Survival / physiology
  • Cells, Cultured
  • Flow Cytometry
  • Ganciclovir / pharmacology
  • Genetic Vectors
  • Herpesviridae / enzymology
  • Humans
  • Immunoenzyme Techniques
  • Liver / immunology
  • Lymphocyte Activation / drug effects
  • Lymphocyte Depletion
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Moloney murine leukemia virus / genetics*
  • Polymerase Chain Reaction
  • Spleen / immunology
  • T-Lymphocytes / physiology*
  • Thymidine Kinase / genetics*
  • Thymus Gland / immunology
  • Transduction, Genetic*


  • Antigens, CD
  • Antiviral Agents
  • Thymidine Kinase
  • Ganciclovir