Advertisement

Non-transplantable cord blood units as a new source for adoptive immunotherapy of leukemia and a paradigm of circular economy in medicine

  • K. Koukoulias
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece

    School of Biology, Department of Genetics, Development and Molecular Biology, Aristotle University, Thessaloniki, Greece
    Search for articles by this author
  • A. Papadopoulou
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece

    School of Biology, Department of Genetics, Development and Molecular Biology, Aristotle University, Thessaloniki, Greece
    Search for articles by this author
  • N. Savvopoulos
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece

    School of Biology, Department of Genetics, Development and Molecular Biology, Aristotle University, Thessaloniki, Greece
    Search for articles by this author
  • E. Deligianni
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece

    School of Biology, Department of Genetics, Development and Molecular Biology, Aristotle University, Thessaloniki, Greece
    Search for articles by this author
  • T. Orfanidou
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece
    Search for articles by this author
  • D. Zervas
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece
    Search for articles by this author
  • A. Xagorari
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece
    Search for articles by this author
  • M. Yiangou
    Affiliations
    School of Biology, Department of Genetics, Development and Molecular Biology, Aristotle University, Thessaloniki, Greece
    Search for articles by this author
  • A. Anagnostopoulos
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece
    Search for articles by this author
  • E. Yannaki
    Affiliations
    Hematology-BMT Unit, Public Cord Blood Bank,Gene and Cell Therapy Center, George Papanicolaou Hospital, Thessaloniki, Greece
    Search for articles by this author
  • P. Kaloyannidis
    Affiliations
    Stemcell Transplant, King Fahad Specialist Hospital, Dammam, Saudi Arabia
    Search for articles by this author
      Immunotherapy with leukemia-specific T cells (leuk-STs) is a promising strategy for leukemia patients lacking a matched donor or relapsing after hematopoietic stem cell transplantation, however, the need for high numbers of antigen presenting cells (APCs) to generate clinically relevant doses of leuk-STs, represents a major challenge. Low volume, non-transplantable cord blood units (CBUs) could potentially serve as a rich and off-the-shelf, indirect source of dendritic cells (DCs) to produce leuk-STs of reduced alloreactivity. We aimed to generate leuk-STs targeting the leukemia-related antigens Wilms tumor protein (WT1) and the Preferentially Expressed Antigen in Melanoma (PRAME), by exploiting disqualified, due to low-volume, CBUs. By culturing 4.2 × 105 ± 1.1 × 105 CD34+ cells from 4 low volume CBUs in Grex devices, in the presence of cytokines (SCF, GM-CSF and IL-4), we generated a median of 3.3 × 109(1.9–5.7 × 109) myeloid DCs (CD33+/CD11c+:76.8 ± 5.5%) in 35 days (fold change~11.000). After exposure to Toll-Like Receptor 3 and 7/8 ligands, the cells expressed maturation markers (CD40+:79 ± 12%, HLA-DR+:78 ± 10%) and secreted high levels of Th1-cytokines (ΙL-12:224 ± 185 pg/ml, IL-6:19 ± 1 × 104 pg/ml, TNF-α:5268 ± 1316 pg/ml) and low levels of the Th2-cytokine, IL-10. Matured DCs were activated with WT1 and PRAME pepmixes and used as APCs to prime previously cryopreserved autologous T-cells derived from the CD34 cell fraction of the CBUs. Expanded cells reached clinically relevant doses (>105/kg) of 7.5 ± 3.4 × 107 CD3+ cells on average, after 4 weeks. Generated cells were polyclonal, expressing memory markers CD45RA+/CD62L:46 ± 4%) while containing insignificant numbers of CD4+/CD25+ cells (1 ± 0.5%). Specificity was seen after the 2nd stimulation at the earliest and was increasing after each stimulation [mean spot forming cells (SFC)/2 × 105 cells at 2nd,3rd,4th stimulation: 106 ± 33; 422 ± 111; 1335 ± 314; respectively]. Produced cells were highly specific for both targeted antigens (PRAME:1019 ± 275, WT1:316 ± 55), while they expressed low the Programmed cell death protein-1 (CD3+/PD-1+:9 ± 4%), implicating absence of cell exhaustion after repeated stimulations. Given the high disqualification rates of CBUs donated to public CB cell banks, we here report a paradigm of “circular economy” in medicine, through the exploitation of non-usable CBUs towards scalable generation of CB-CD34+cell-derived DCs and CB-CD34cell-derived leuk-STs and establishment of third-party leuk-STs banks.
      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Cytotherapy
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect