Homing and survival of thymidine kinase-transduced human T cells in NOD/SCID mice.
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.
Hematology and Clinical Immunology Section, Department of Clinical and Experimental Medicine, Perugia University, Perugia, Italy.
Mauro
M
Di Ianni
Adelmo
A
Terenzi
Franca
F
Falzetti
Andrea
A
Bartoli
Sabrina
S
Di Florio
Roberta
R
Benedetti
Gigliola
G
Venditti
Diego
D
Alfonsi
Mariangela
M
De Ioanni
Brunangelo
B
Falini
Antonio
A
Tabilio
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D000998:Antiviral Agents / Q000494:pharmacology; D001853:Bone Marrow / Q000276:immunology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D005434:Flow Cytometry; D015774:Ganciclovir / Q000494:pharmacology; D005822:Genetic Vectors; D006564:Herpesviridae / Q000201:enzymology; D006801:Humans; D007124:Immunoenzyme Techniques; D008099:Liver / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects; D008212:Lymphocyte Depletion; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008979:Moloney murine leukemia virus / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013154:Spleen / Q000276:immunology; D013601:T-Lymphocytes / Q000502:physiology*; D013937:Thymidine Kinase / Q000235:genetics*; D013950:Thymus Gland / Q000276:immunology; D014161:Transduction, Genetic
D015703:Antigens, CD; D000998:Antiviral Agents; D013937:Thymidine Kinase; D015774:Ganciclovir