Natural Killer Cell Protocols

Research has revealed that natural killer (NK) cells are sentinels of the innate immune response. This text offers detailed protocols plus the latest methods and techniques designed to study NK cells within specialized tissue sites in both mice and humans.

As the research has continued, it has become increasingly clear that natural killer (NK) cells are critical sentinels of the innate immune response, playing important roles in protecting the body from numerous pathogens and cancer in addition to contributing to normal pregnancy and impacting the outcomes of transplantation. While the first edition provided a valuable collection of classical cellular and in vivo techniques to study NK cell functions, the Second Edition of "Natural Killer Cell Protocols: Cellular and Molecular Methods" brings together more recently developed methods, more refined techniques, and detailed protocols designed to study NK cells within specialized tissue sites in both mice and humans. In this collection of methods, international leaders in the field cover topics ranging from the analysis of the various stages of NK cell development and maturation to specialized techniques for the identification of ligands for NK cell receptors. This volume also includes an appendix, providing a rich resource summarizing available reagents to study NK cells, cross-referencing KIR nomenclature, and detailing the many HLA ligands for various KIR family members. As a volume in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and thorough notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, "Natural Killer Cell Protocols: Cellular and Molecular Methods, Second Edition" seeks to aid researchers and further advance our understanding of the functions, maturation, and regulation of these fascinating and dynamic cells.

Remains the most comprehensive collection of methods available for studying natural killer cells in both mice and humans Examines a wide array of molecular and cellular techniques for manipulating and assaying NK cell functions both in vitro and in vivo Includes techniques to assay NK cell phenotype, development, signaling, and functional responses to tumor cells, virus infection, and malaria infection Contains a wide array of techniques to assay NK cells by flow cytometry Provides molecular techniques to assay important genotypes, map functionally important genetic loci in mice, and identify genetic regulatory elements Supplies several specialized protocols to identify novel ligands for NK cell receptors Includes specialized techniques for studying NK cells from tonsils, lymph nodes, uterine tissues, and gut mucosa Includes clinically relevant assays of human killer cell Ig-like receptor (KIR) genotype and surface expression phenotype by flow cytometry, as well as detailed information on KIR nomenclature and ligand specificities Highlights information on nomenclature, cell lines, available antibodies, and receptor ligands in its valuable appendices Includes supplementary material: sn.pub/extras

Inhalt
Purification of Human NK Cell Developmental Intermediates from Lymph Nodes and Tonsils.- In Vitro Development of Human KillerImmunoglobulin Receptor-Positive NK Cells.- Subset Analysis of Human and Mouse Mature NK Cells.- Assessing Licensing of NK Cells.- Use of Stem Cell Radiation Chimeras to Analyze How Domains of Specific Proteins Impact on Murine NK Cell Development In Vivo.- Use of Transfected Drosophila S2 Cells to Study NK Cell Activation.- Natural Killer Cell Conjugate Assay Using Two-Color Flow Cytometry.- Studying NK Cell/Dendritic Cell Interactions.- Analysis of the NK Cell Immunological Synapse.- Measuring Intracellular Calcium Signaling in Murine NK Cells by Flow Cytometry.- Intracellular Staining for Analysis of the Expression and Phosphorylation of Signal Transducers and Activators of Transcription (STATs) in NK Cells.- A Model System for Studying NK Cell Receptor Signaling.- Expression of cDNAs in Human Natural Killer Cell Lines by Retroviral Transduction.- Lentiviral Gene Transduction in Human and Mouse NK Cell Lines.- of shRNAs into Human NK-Like Cell Lines with Retrovirus.- of shRNAs into Primary NK Cells with Lentivirus.- Methods to Identify and Characterize Different NK Cell Receptors and Their Ligands.- Generating NK Cell Receptor-Fc Chimera Proteins from 293T Cells and Considerations of Appropriate Glycosylation.- Identification of NK Cell Receptor Ligands Using a Signaling Reporter System.- Determining Ligand Specificity of Ly49 Receptors.- Probing the Interactions of NK Cell Receptors with Ligand Expressed in trans and cis.- A Simple Method to Measure NK Cell Cytotoxicity In Vivo.- Functional Analysis of Human NK Cells by Flow Cytometry.- Analysis of the KIR Repertoire in Human NK Cells by Flow Cytometry.- KIR Genotyping by MultiplexPCR-SSP.- Identification and Analysis of Novel Transcripts and Promoters in the Human Killer Cell Immunoglobulin-like Receptor (KIR?) Genes.- Use of Inbred Mouse Strains to Map Recognition Receptors of MCMV Infected Cells in the NK Cell Gene Locus.- Studying NK Cell Responses to Ectromelia Virus Infections in Mice.- Activation of Human NK Cells by Malaria-Infected Red Blood Cells.- Natural Killer Cells in Human Pregnancy.- Analysis of Uterine Natural Killer Cells in Mice.- Isolation of NK Cells and NK-Like Cells from the Intestinal Lamina Propria.