Improving Clinical Manufacturing of IL-15 Activated Cytokine-Induced Killer (CIK) Cells

Abstract

Cytokine-induced killer (CIK) cells are an immunotherapeutic approach to combat relapse following allogeneic hematopoietic stem cell transplantation (HSCT) in acute leukemia or myelodysplastic syndrome (MDS) patients. Prompt and sequential administration of escalating cell doses improves the efficacy of CIK cell therapy without exacerbating graft vs. host disease (GVHD). This study addresses manufacturing-related issues and aimed to develop a time-, personal- and cost-saving good manufacturing process (GMP)-compliant protocol for the generation of ready-for-use therapeutic CIK cell doses starting from one unstimulated donor-derived peripheral blood (PB) or leukocytapheresis (LP) products. Culture medium with or without the addition of either AB serum, fresh frozen plasma (FFP) or platelet lysate (PL) was used for culture. Fresh and cryopreserved CIK cells were compared regarding expansion rate, viability, phenotype, and ability to inhibit leukemia growth. Cell numbers increased by a median factor of 10-fold in the presence of FFP, PL, or AB serum, whereas cultivation in FFP/PL-free or AB serum-free medium failed to promote adequate CIK cell proliferation ( p < 0.01) needed to provide clinical doses of 1 × 10 6 T cells/kG, 5 × 10 6 T cells/kG, 1 × 10 7 T cells/kG, and 1 × 10 8 T cells/kG recipient body weight. CIK cells consisting of T cells, T- natural killer (T-NK) cells and a minor fraction of NK cells were not significantly modified by different medium supplements. Moreover, neither cytotoxic potential against leukemic THP-1 cells nor cell activation shown by CD25 expression were significantly influenced. Moreover, overnight and long-term cryopreservation had no significant effect on the composition of CIK cells, their phenotype or cytotoxic potential. A viability of almost 93% (range: 89–96) and 89.3% (range: 84–94) was obtained after freeze-thawing procedure and long-term storage, respectively, whereas viability was 96% (range: 90-97) in fresh CIK cells. Altogether, GMP-complaint CIK cell generation from an unstimulated donor-derived PB or LP products was feasible. Introducing FFP, which is easily accessible, into CIK cell cultures was time- and cost-saving without loss of viability and potency in a 10-12 day batch culture. The feasibility of cryopreservation enabled storage and delivery of sequential highly effective ready-for-use CIK cell doses and therefore reduced the number of manufacturing cycles.

Adoptive cell therapy of triple negative breast cancer with redirected cytokine-induced killer cells

“…CD25 is the IL-2 receptor α-chain that interacts with the IL-2Rβ and IL-2Rγ subunits to form the high-affinity IL-2 receptor; 30 CD25 overexpression may facilitate the efficacy of cell therapies through the creation of an extracellular reservoir and recycling of IL-2, 31 and its upregulation correlates with the cytotoxic activity in NK and CIK cells. 32,33 Here we demonstrated that a CD25 upregulation occurred exclusively in the CD3 + CD56 + CD16 + subset and only in the presence of the antigen-specific mAb that could explain the enrichment of the CD3 + CD56 + CD16 + fraction within both primary and metastatic tumors when CTX was used.…”

ERBB2-CAR-Engineered Cytokine-Induced Killer Cells Exhibit Both CAR-Mediated and Innate Immunity Against High-Risk Rhabdomyosarcoma

“…In previous work, we employed CIK cells after ex vivo activation with IL-15 for 10 days, which resulted in more rapid generation of CIK cells with stronger cytotoxicity compared to conventional CIK cells expanded in the absence of IL-15 over 3-4 weeks. (32,46,47). The main fraction in this short-term, IL-15-activated CIK cell culture is the CD3 + T cell population.…”

Killers at the crossroads: The use of innate immune cells in adoptive cellular therapy of cancer

“… 59‐61 Several protocols for the generation of CIK cells have been reported, and typically involve the sequential incubation of peripheral blood lymphocytes with interferon (IFN)‐γ, anti‐CD3 mAb, and IL‐2 62 . The use of IL‐15 in the presence or absence of IL‐2 has also been shown to induce faster expansion of CIK cells to exhibit enhanced cytotoxicity, with the additional advantage of inducing fewer numbers of suppressive Treg cells 63,64 . CIK cells exhibit stronger cytolytic activity compared to lymphokine‐activated killer (LAK) cells, which are generated by incubation with interleukins.…”

“…CIK cells exhibit stronger cytolytic activity compared to lymphokine‐activated killer (LAK) cells, which are generated by incubation with interleukins. Enhanced function of CIK cells relative to LAK cells has been attributed to a higher proliferative capacity and an increase in lytic units 64,65 …”