Generation of functional CLL-specific cord blood CTL using CD40-ligated CLL APC

PLoS One. 2012;7(12):e51390. doi: 10.1371/journal.pone.0051390. Epub 2012 Dec 19.


Though remissions have been observed following allo-HSCT for the treatment of CLL, many CLL patients are ineligible for transplant due to the lack of HLA-compatible donors. The use of umbilical cord blood (UCB) permits transplantation of many patients who lack HLA-compatible donors due to reduced requirements for stringent HLA matching between graft and recipient; however, disease relapse remains a concern with this modality. The generation of CLL-specific CTL from UCB T-cells, primed and expanded against the leukemic clone, might enhance the GVL effect and improve outcomes with UCB transplantation. Here we report the generation of functional, CLL-specific CTL using CD40-ligated CLL cells to prime partially-HLA matched UCB T-cells. Functionality and specificity were demonstrated by immune synapse assay, IFN-γ ELISpot, multi-parametric intracellular cytokine flow cytometry, and (51)Cr release assay. The use of patient-specific, non-CLL controls demonstrated the generation of both alloantigen and CLL-specific responses. Subsequently, we developed a clinically-applicable procedure permitting separation of alloreactive CTL from leukemia-specific CTL. Leukemia-specific CTL were able to mediate in vivo killing of CLL in humanized mice without concurrent or subsequent development of xenoGVHD. Our results demonstrate that generation of CLL-specific effectors from UCB is feasible and practical, and the results support further exploration of this strategy as a treatment modality for CLL.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Animals
  • Antigen-Presenting Cells / immunology*
  • Antigen-Presenting Cells / metabolism*
  • Antigens, Neoplasm / immunology
  • CD40 Antigens / immunology
  • CD40 Antigens / metabolism*
  • Feasibility Studies
  • Fetal Blood / immunology*
  • HLA Antigens / immunology
  • Humans
  • Immunological Synapses / immunology
  • Leukemia, Lymphocytic, Chronic, B-Cell / immunology*
  • Mice
  • T-Lymphocytes / immunology


  • Antigens, Neoplasm
  • CD40 Antigens
  • HLA Antigens