Single-cell cloning of human, donor-derived antileukemia T-Cell lines for in vitro separation of graft-versus-leukemia effect from graft-versus-host reaction

Department of Pediatrics, Laboratory of Immunology, University of Pavia, Italy.
Cancer Research (Impact Factor: 9.33). 08/2006; 66(14):7310-6. DOI: 10.1158/0008-5472.CAN-06-0591
Source: PubMed


In previous studies, we showed the possibility of expanding in vitro polyclonal CTL lines directed against patient leukemia cells using effector cells derived from both HLA-matched and HLA-mismatched hematopoietic stem cell donors. Some CTL lines, especially those derived from an HLA-disparate donor, displayed residual alloreactivity against patient nonmalignant cells. In this study, we evaluated the possibility of separating in vitro CTLs with selective graft-versus-leukemia (GVL) activity from those potentially involved in the development of graft-versus-host disease (GVHD) through single T-cell cloning of antileukemia polyclonal CTL lines. We showed that CTLs that were expanded from a single T-cell clone (TCC), able to selectively kill leukemia blasts and devoid of alloreactivity towards nonmalignant cells, can be obtained from antileukemia alloreactive polyclonal CTL lines. TCCs expressed a wide repertoire of different T-cell receptor (TCR)-Vbeta families, mainly produced IFNgamma and interleukin 2, irrespective of CD8 or CD4 phenotype, and could be extensively expanded in vitro without losing their peculiar functional features. The feasibility of our approach for in vitro separation of GVL from GVH reaction opens perspectives for using TCCs, which are selectively reactive towards leukemia blasts, for antileukemia adoptive immune therapy approaches after hematopoietic stem cell transplantation, in particular from HLA-mismatched donors.

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Available from: Maria Ester Bernardo, Jun 24, 2014
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    • "For HSCT recipients from HLA-matched donors, the GVL effect can be triggered by minor histocompatibility antigens [2] [3] [4], and several studies using sequential flow cytometric analysis with tetramers have clearly demonstrated that minor histocompatibility antigen-specific T lymphocytes increase in frequency in the recipient's blood before and during clinical regression of leukemia [5–10]. On the other hand, for HLA-mismatched HSCT recipients , extremely limited biological studies have demonstrated that the GVL effect can be mediated by mismatched HLA-specific donor T lymphocytes [11]. Allogeneic HSCT is a well-established immunotherapy for leukemia, but, unfortunately, some recipients relapse after transplantation . "
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