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Cryopreservation of Natural Killer Cells Pre-Complexed with Innate Cell Engagers.

Uwe Reusch1, Kristina Ellwanger1, Ivica Fucek1

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Antibodies (Basel, Switzerland)
|February 28, 2022
PubMed
Summary

Innate cell engager (ICE) constructs pre-complexed with natural killer (NK) cells demonstrate durable binding and potent anti-tumor activity. These ICE-NK cell complexes are suitable for cryopreservation, offering potential for off-the-shelf cell therapies.

Keywords:
adoptive transferbispecific antibodycryopreservationinnate cell engagerinnate immune cellsnatural killer cell

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Area of Science:

  • Immunology
  • Oncology
  • Biotechnology

Background:

  • Innate cell engager (ICE®) constructs are bispecific antibodies designed to link tumor cells with immune effector cells like NK cells and macrophages.
  • Pre-complexing ICE® constructs with adoptive NK cells represents a novel strategy to enhance anti-tumor immune responses.
  • The development of cryopreserved, off-the-shelf NK cell products is a significant goal in cancer immunotherapy.

Purpose of the Study:

  • To investigate the binding affinity and anti-tumor efficacy of ICE® constructs targeting EpCAM and NK cell receptors (CD16A, NKG2D, NKp46).
  • To assess the feasibility of cryopreserving ICE®-NK cell complexes while maintaining biological activity.
  • To evaluate the potential of these complexes as off-the-shelf NK cell therapies.

Main Methods:

  • Cell surface retention assays using primary NK cells to determine ICE® construct dissociation kinetics.
  • Evaluation of ICE® construct anti-tumor activity against EpCAM-expressing tumor cells.
  • Assessment of the impact of cryopreservation (freeze-thaw cycles) on the potency and efficacy of ICE®-NK cell complexes.

Main Results:

  • ICE® constructs exhibited significantly slower dissociation from NK cells compared to control molecules, indicating durable complex formation.
  • High-affinity ICE® complexes targeting EpCAM and CD16A demonstrated superior efficacy in eliminating tumor cells compared to those engaging NKG2D or NKp46.
  • The potency and efficacy of the ICE®-NK cell complexes remained unaffected after a single freeze-thaw cycle.

Conclusions:

  • CD16A-selective ICE® constructs complexed with NK cells show promise as novel cryopreserved, off-the-shelf products.
  • These complexes possess chimeric antigen receptor-like NK cell properties, enabling effective tumor cell depletion.
  • The findings support the development of readily available NK cell-based immunotherapies.