Work

Adoptive Immunotherapy After Umbilical Cord Blood (UCB) Transplantation: Manufacturing And Analysis Of CD19-Specific UCB-Derived T-cells From Scant Numbers Of UCB Mononuclear Cells

Year: 2010

Type: article

Abstract: The rapid procurement, lower risk of graft-versus-host-disease (GvHD), and the ability to undertake transplantation across HLA disparities make umbilical cord blood (UCB) an attractive stem-cell source. Adoptive transfer of tumor-specific T cells after UCB transplantation (UCBT) is constrained by the functionally naïve UCB T cells and lack of ability to obtain additional cells. We have previously shown that the specificity of T cells can be redirected for CD19 using a chimeric antigen receptor (CAR) independent of major histocompatibility complex. We have identified a source of clinical-grade UCB that may be used for manufacturing without having to reduce the cell dose available for transplantation. In our laboratory, after UCB units are washed for clinical infusion, the supernatant is refrozen, and contains small numbers of UCB cells, including T cells. We evaluated the feasibility of generating UCB-derived CD19-specific T cells for clinical infusion from the T cells in this residual. These T cells were electroporated with CD19RCD28 CAR expressed as a Sleeping Beauty (SB) transposon and propagated in the presence of IL-2 and IL-21 on g-irradiated CD19+ K562 artificial antigen presenting cells (aAPC). The electroporated and propagated cells were a population of CD8+ and CD4+ memory and effector T cells that exhibit redirected CD19-dependent killing and cytokine production. The UCB-derived CD19-specific T cells were further analyzed using a new technology using the NanoString nCounter Analysis System, a digital technology that is based on direct multiplexed measurement of mRNA and therefore gene expression that offers a high level of precision and sensitivity from a few number of cells. This approach to manufacturing CD19-specific T cells and analysis of mRNA expression profiles will be used to generate donor-derived CD19-specific T cells that can be infused after UCBT as an approach to enhance the graft-versus-tumor-effect without increasing GvHD. The relative ease and low cost associated with production of clinical grade DNA plasmids, electroporation, and reproducible outgrowth of T cells stablely expressing CAR on thawed g-irradiated MCB of aAPC, should enable facilities operating in compliance with cGMP for Phase I/II to implement adoptive immunotherapy trials using CAR+ T cells. By using the scant numbers of residual UCB cells present after washing, we have identified one potential way to overcome a major barrier to UCB-derived T-cell therapies.

Source: Transplantation and Cellular Therapy

DOI: 10.1016/j.bbmt.2009.12.097