Reduced graft-versus-host disease-inducing capacity of T cells after activation, culturing, and magnetic cell sorting selection in an allogeneic bone marrow transplantation model in rats.
Graft-versus-host disease (GvHD), a major complication of allogeneic bone marrow transplantation, has been ascribed to mature T cells in the graft. Because T cells play an important role in engraftment of the bone marrow and decrease the probability of relapse of leukemia, a treatment strategy was developed to preserve the benefits of T cells in the graft and to control the severe complications of GvHD. This can be accomplished by the genetic modification of donor T cells with a suicide gene that allows their selective in vivo elimination and subsequently the abrogation of GvHD. For clinical benefit the alloreactivity of herpes simplex virus thymidine kinase (HSV-TK) gene-transduced T cells should be retained. Therefore, we investigated the influence of gene transduction and the selection procedure on T cells. We demonstrated that activation and culturing of T cells reduce their capacity to induce lethal GvHD in an allogeneic rat bone marrow transplantation model. Furthermore, positive immunomagnetic selection of gene-transduced T cells resulted in loss of the GvHD-inducing capacity of HSV-TK(+) T cells directly after MACS (magnetic cell sorting) selection; this loss could be recovered by a 1-day expansion of the selected T cells. No effect on alloreactivity was observed to be caused by the gene transduction procedure. Our study resulted in the development of an optimized culture and gene transduction protocol with preservation of T cell alloreactivity. Treatment of transplanted rats with ganciclovir resulted in a rapid reduction in the number of HSV-TK(+) T cells in the peripheral blood and in increased survival of the animals.
Weijtens
Mo
M
Jordan Laboratory of Hemato-Oncology, Department of Hematology, University Medical Center, 3584 CX Utrecht, The Netherlands.
van Spronsen
Anke
A
Hagenbeek
Anton
A
Braakman
Eric
E
Martens
Anton
A
D000818:Animals; D001835:Body Weight; D016026:Bone Marrow Transplantation / Q000009:adverse effects / Q000276:immunology*; D018929:Cell Culture Techniques; D002460:Cell Line; D004195:Disease Models, Animal; D015774:Ganciclovir / Q000627:therapeutic use; D006086:Graft vs Host Disease / Q000276:immunology* / Q000517:prevention & control; D018189:Immunomagnetic Separation; D008213:Lymphocyte Activation; D051381:Rats; D011914:Rats, Inbred BN; D018139:Simplexvirus / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*; D013937:Thymidine Kinase / Q000235:genetics; D014161:Transduction, Genetic
D013937:Thymidine Kinase; D015774:Ganciclovir