Innate Lymphoid Cells Methods and Protocols

This volume explores the latest techniques used to characterize murine and human Innate Lymphoid Cells (ILC) subsets. This book also aims to discuss useful phenotypical and functional assays that may be relevant for studying ILC function in mice and humans. Chapters cover topics such as methods to study microbiome and enteroid cultures; characterizing ILC precursors; and cellular engineering of ILCs for therapeutic use. Written 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 tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and practical, Innate Lymphoid Cells: Methods and Protocols is a valuable research tool for anyone interested in ILC biology, and those interested in fundamental immunology, cellular engineering, cell signaling, and clinician scientists. 

Acknowledgements…
Preface…
Table of Contents…
Contributing Authors…

1. Protocols for Innate Lymphoid Cell Phenotypic and Functional Characterization: An Overview
Shoba Amarnath

2. Protocols for Studying Murine ILC Development 
Matthew T. Stier and R. Stokes Peebles, Jr.

3. Identification of Murine Innate Lymphoid Cell Subsets in Barrier Tissues and their Draining Lymph Nodes using Flow Cytometry
Emma E. Dutton and David R. Withers

4. Identification and Functional Characterization of Murine Group 3 Innate Lymphoid Cell (ILC3) Subsets in the Intestinal Tract and Associated Lymphoid Tissues
Felipe Melo-Gonzalez and Matthew R. Hepworth

5. Identification of Murine and Human Innate Lymphoid Cells in Frozen Tissue Sections using Immunofluorescence
Emma E. Dutton and David R. Withers

6. Assessing Phosphorylation of STAT Transcription Factors in Mouse Innate Lymphoid Cells
Gianluca Scarno, Giuseppe Pietropaolo, Chiara Di Censo, Giovanna Peruzzi, and Giuseppe Sciumè

7. Determining Co-Receptor Expression and Function in Murine ILC2 through Flow Cytometry Characterization and Co-Culture Techniques
Christian Schwartz and Padraic G. Fallon

8. Protocol for Determining the Effect of Neuroendocrine Hormones on Murine ILC Function
Linda Quatrini and Sophie Ugolini

9. Role of G-Protein Coupled Receptors in Chemotaxis of Innate Lymphoid Cells
Brinda Bhatt, Huabin Zhu, Mikhil Patel, and Magendra Singh

10. Group 2 ILC Functional Assays in Allergic Airway Inflammation
Lewis J. Entwistle, Franz Puttur, Lisa G. Gregory, and Clare M. Lloyd

11. Isolation, Culture, and Induction of Plasticity in ILC2s
Jonathan Silver, Alison A. Humbles, and Yoichiro Ohne

12. Bacterial Infection Allows for Functional Examination of Adoptively Transferred Mouse Innate Lymphoid Cell Subsets
Goo-Young Seo, Daniel A. Giles, and Mitchell Kronenberg

13. Isolation, Characterization, and Culture of Intestinal Intraepithelial Lymphocytes
Olivia Jane James, Maud Vandereyken, and Mahima Swamy

14. Isolation and Characterization of Innate Lymphoid Cells within the Murine Tumor Microenvironment
Grace Mallett, Warren Patterson, Megan Payne, and Shoba Amarnath

15. Microbiota Analysis using Sequencing by Synthesis: From Library Preparation to Sequencing
Andrew Nelson and Christopher J. Stewart

16. Establishing Human Intestinal Enteroid/Organoid Lines from Preterm Infant and Adult Tissue
Christopher J. Stewart, Mary K. Estes, and Sasirekha Ramani

17. Characterizing Innate Lymphoid Cell Phenotype and Function in Human Inflammatory Bowel Disease
Omer S. Omer, Nick Powell, and Graham M. Lord

18. CRISPR/Cas9-Based Gene Engineering of Human Natural Killer Cells: Protocols for Knock-Out and Read-Outs to Evaluate the Efficacy
Melanie Lambert, Caroline Leijonhufvud, Filip Segerberg, J. Joseph Melenhorst, and Mattias Carlsten

Subject Index List…