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Modification of Expanded NK Cells with Chimeric Antigen Receptor mRNA for Adoptive Cellular Therapy

  • Yaya Chu
  • Allyson Flower
  • Mitchell S. Cairo
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1441)

Abstract

NK cells are bone marrow-derived cytotoxic lymphocytes that play a major role in the rejection of tumors and cells infected by viruses. The regulation of NK activation vs inhibition is regulated by the expression of a variety of NK receptors (NKRs) and specific NKRs’ ligands expressed on their targets. However, factors limiting NK therapy include small numbers of active NK cells in unexpanded peripheral blood and lack of specific tumor targeting. Chimeric antigen receptors (CAR) usually include a single-chain Fv variable fragment from a monoclonal antibody, a transmembrane hinge region, and a signaling domain such as CD28, CD3-zeta, 4-1BB (CD137), or 2B4 (CD244) endodimers. Redirecting NK cells with a CAR will circumvent the limitations of the lack of NK targeting specificity. This chapter focuses on the methods to expand human NK cells from peripheral blood by co-culturing with feeder cells and to modify the expanded NK cells efficiently with the in vitro transcribed CAR mRNA by electroporation and to test the functionality of the CAR-modified expanded NK cells for use in adoptive cellular immunotherapy.

Key words

Chimeric antigen receptor Natural killer cells mRNA Electroporation Adoptive cell therapy 

Notes

Acknowledgements

The authors thank Erin Morris, RN, for her excellent assistance with the preparation of this manuscript. The authors also thank Dr. Dario Campana (St. Jude Children’s Research Hospital) and Dr. Terrence Geiger (St. Jude Children’s Research Hospital) for kindly providing anti-CD20 scFv. The research for this study was supported by the grant from the Pediatric Cancer Research Foundation and the New York Medical College Intramural Research Award.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Yaya Chu
    • 1
  • Allyson Flower
    • 1
  • Mitchell S. Cairo
    • 1
    • 2
    • 3
    • 4
    • 5
    • 6
  1. 1.Department of PediatricsNew York Medical CollegeValhallaUSA
  2. 2.Department of MedicineMaria Fareri Children’s Hospital at Westchester Medical Center, New York Medical CollegeValhallaUSA
  3. 3.Department of PathologyNew York Medical CollegeValhallaUSA
  4. 4.Department of Microbiology and ImmunologyNew York Medical CollegeValhallaUSA
  5. 5.Department of Cell Biology and AnatomyNew York Medical CollegeValhallaUSA
  6. 6.Maria Fareri Children’s Hospital at Westchester Medical CenterNew York Medical CollegeValhallaUSA

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