Main description:

This volume contains collection of Natural Killer Cell methodologies relevant for both basic and translational research. These methodologies present new developments in the natural killer (NK) cell field, such as understanding the influence of NK cells metabolism on its function, identifying complexity of NK cell subsets through mass cytometry, and determining the emergence of memory NK cells in murine model of MCMV infection. Methods that study NK cell migration and cytotoxicity through endpoint analysis or live single cell imaging are also discussed. Chapters also describe methods pertaining to translational application of NK cells, such as ex vivo expansion of NK cells on K562 cell lines genetically modified to express either membrane bound IL-15 or membrane bound IL-21, large scale NK cell culture, current techniques for engineering NK cells to express chimeric antigen receptors or chemokine receptors using retroviral vectors, electroporation of mRNA, and the natural phenomenon of trogocytosis. Written in the highly successful Methods in Molecular Biology series format, these chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting edge and thorough, Natural Killer Cells: Methods and Protocols is a valuable resource for researchers who not only want to understand mechanisms that govern NK cell behavior and diversity, but also for those who want to understand how to systematically evaluate NK cells for adoptive immunotherapy applications.

Contents:

1. Tracking
Effector and Memory NK Cells During MCMV Infection
Aimee
M. Beaulieu and Joseph C. Sun

2. Application
of Mass Cytometry (CyTOF) for Functional and Phenotypic Analysis of Natural Killer
Cells
Alexander W. Kay, Dara M.
Strauss-Albee, and Catherine A. Blish

3. Assessment
of NK Cell Metabolism
Molly P. Keppel and Megan A.
Cooper

4. Genotyping Single Nucleotide Polymorphisms and
Copy Number Variability of the FCGRs Expressed on NK Cells
Amy K. Erbe, Wei Wang, Mikayla
Gallenberger, Jacquelyn A. Hank, and Paul M. Sondel
5. Measurement
of Average Telomere Length in Ex Vivo
Expanded Natural Killer Cells by Fluorescence In Situ Hybridization (FISH)
Sourindra N. Maiti
6. In
Vitro Assessment of Human Natural Killer Cell Migration and Invasion
Karin Tomin, Ronald H. Goldfarb,
and Per Albertsson
7. Microfluidic-Based
Live-Cell Analysis of NK-Cell Migration In Vitro
Saravanan Nandagopal, Francis Lin, and Sam
K.P. Kung
8. Microwell-Based
Live Cell Imaging of NK Cell Dynamics to Assess Heterogeneity in Motility and Cytotoxic
Response
Bruno Vanherberghen, Thomas Frisk, Elin Forslund, Per E. Olofsson, Karolin
Guldevall, and Bjoern OEnfelt
9. Assessment
of Natural Killer Cell Cytotoxicity Using Image Cytometry Method
Kelsey
J. McCulley and Srinivas S. Somanchi
10. Analysis
of Intracellular Ca2+ Mobilization in Human NK Cell Subsets by Flow Cytometry
Jakob
Theorell and Yenan T. Bryceson
11. Using NK Cell Lipid Raft Fractionation to Understand
the Role of Lipid Rafts in NK Cell Receptor SignalingEsther Serrano-Pertierra and Carlos Lopez-Larrea
12. High-
and Super-Resolution Microscopy Imaging of the NK Cell Immunological Synapse
Emily M. Mace and Jordan S.
Orange
13. The
Planar Lipid Bilayer System Serves as a Reductionist Approach for Studying NK
Cell Immunological Synapses and their Functions
Grant Bertolet and Dongfang Liu
14. Expansion
of NK Cells Using Genetically Engineered K562 Feeder Cells
Minh-Trang Thi Phan, Seung-hwan Lee, Sang-Ki Kim, and Duck
Cho

15. Ex-Vivo
Expansion of Human NK Cells Using K562 Engineered to Express Membrane Bound
IL21
Srinivas
S. Somanchi and Dean A. Lee

16. Large-Scale
Culture and Genetic Modification of Human Natural Killer Cells for Cellular
TherapyNatalia
Lapteva, Robin Parihar, Lisa A. Rollins, Adrian P. Gee, and Cliona M. Rooney
17. Gene
Modification of Human Natural Killer Cells Using a Retroviral VectorJoshua N. Kellner, Conrad R.
Cruz, Catherine M. Bollard, and Eric S. Yvon
18. Modification
of Expanded NK Cells with Chimeric Antigen Receptor mRNA for Adoptive Cellular
TherapyYaya
Chu, Allyson Flower, and Mitchell S. Cairo
19. mRNA
Transfection to Improve NK Cell Homing to TumorsEmily
Levy, Mattias Carlsten, and Richard Childs
20. In
Vitro Generation of Human NK Cells Expressing Chimeric Antigen Receptor through
Differentiation of Gene-Modified Hematopoietic Stem CellsEmily
Lowe, Laurel C. Truscott, and Satiro N. De Oliveira
21. Engineering
Receptor Expression on Natural Killer Cells through TrogocytosisAnitha
Somanchi, Dean A. Lee, and Srinivas S. Somanchi
22. Electroporation
of siRNA to Silence Gene Expression in Primary NK CellsPrasad V. Phatarpekar, Dean A.
Lee, and Srinivas S. Somanchi
23.
Mouse Xenograft Model for Intraperitoneal Administration
of NK Cell Immunotherapy for Ovarian CancerDavid
L. Hermanson, Laura Bendzick, and Dan S. Kaufman
24. Aerosol
Delivery of Interleukin-2 in Combination with Adoptive Transfer of Natural
Killer Cells for the Treatment of Lung Metastasis: Methodology and EffectSimin
Kiany and Nancy Gordon
25. Non-Invasive Imaging of Natural Killer Cell-Mediated
Apoptosis in a Mouse Tumor Model Thoudam
Debraj Singh, Jaetae Lee, and Yong Hyun Jeon
26. Non-Invasive
In Vivo Fluorescence Imaging of NK Cells
in Pre-Clinical Models of Adoptive ImmunotherapySrinivas
S. Somanchi
27. In Vivo 19F-Magnetic
Resonance Imaging of Adoptively Transferred NK CellsSrinivas
S. Somanchi, Bridget A. Kennis, Vidya Gopalakrishnan, Dean A. Lee, and James A.
Bankson
28. Generation
of BiKEs and TriKEs to Improve NK Cell-Mediated Targeting of Tumor CellsMartin Felices, Todd R. Lenvik,
Zachary B. Davis, Jeffrey S. Miller, and Daniel A. Vallera
29. Regulatory
Considerations for NK Cells Used in Human Immunotherapy ApplicationsDean A. Lee