Natural Killer Cells

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.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Tracking Effector and Memory NK Cells During MCMV Infection.- Application of Mass Cytometry (CyTOF) for Functional and Phenotypic Analysis of Natural Killer Cells.- Assessment of NK Cell Metabolism.- Genotyping Single Nucleotide Polymorphisms and Copy Number Variability of the FCGRs Expressed on NK Cells.- Measurement of Average Telomere Length in Ex Vivo Expanded Natural Killer Cells by Fluorescence In Situ Hybridization (FISH).- In Vitro Assessment of Human Natural Killer Cell Migration and Invasion.- Microfluidic-Based Live-Cell Analysis of NK-Cell Migration In Vitro.- Microwell-Based Live Cell Imaging of NK Cell Dynamics to Assess Heterogeneity in Motility and Cytotoxic Response.- Assessment of Natural Killer Cell Cytotoxicity Using Image Cytometry Method.- Analysis of Intracellular Ca 2+ Mobilization in Human NK Cell Subsets by Flow Cytometry.- Using NK Cell Lipid Raft Fractionation to Understand the Role of Lipid Rafts in NK Cell Receptor Signaling.- High- and Super-Resolution Microscopy Imaging of the NK Cell Immunological Synapse.- The Planar Lipid Bilayer System Serves as a Reductionist Approach for Studying NK Cell Immunological Synapses and their Functions.- Expansion of NK Cells Using Genetically Engineered K562 Feeder Cells.- Ex-Vivo Expansion of Human NK Cells Using K562 Engineered to Express Membrane Bound IL21.- Large-Scale Culture and Genetic Modification of Human Natural Killer Cells for Cellular Therapy.- Gene Modification of Human Natural Killer Cells Using a Retroviral Vector.- Modification of Expanded NK Cells with Chimeric Antigen Receptor mRNA for Adoptive Cellular Therapy.- mRNA Transfection to Improve NK Cell Homing to Tumors.- In Vitro Generation of Human NK Cells Expressing Chimeric Antigen Receptor through Differentiation of Gene-Modified Hematopoietic Stem Cells.- Engineering Receptor Expression on Natural Killer Cells through Trogocytosis.- Electroporation of siRNA to Silence Gene Expression in Primary NK Cells.- Mouse Xenograft Model for Intraperitoneal Administration of NK Cell Immunotherapy for Ovarian Cancer.- Aerosol Delivery of Interleukin-2 in Combination with Adoptive Transfer of Natural Killer Cells for the Treatment of Lung Metastasis: Methodology and Effect.- Non-Invasive Imaging of Natural Killer Cell-Mediated Apoptosis in a Mouse Tumor Model.- Non-Invasive In Vivo Fluorescence Imaging of NK Cells in Pre-Clinical Models of Adoptive Immunotherapy.- In Vivo 19 F-Magnetic Resonance Imaging of Adoptively Transferred NK Cells.- Generation of BiKEs and TriKEs to Improve NK Cell-Mediated Targeting of Tumor Cells.- Regulatory Considerations for NK Cells Used in Human Immunotherapy Applications.