Type 2 Immunity, 1st ed. 2018
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1799

This book provides researchers the opportunity to investigate type-2-associated diseases in their laboratories. Beginning with chapters describing various models of type-2 immunity, the volume then continues by detailing cellular protocols designed to identify, characterize, and assess the function of key adaptive and innate immune cells involved in type-2 inflammation; approaches to isolate and evaluate specific cellular subsets at the genetic, epigenetic, and molecular level; protocols to assess type-2 immunity and its relationship to organismal and metabolic systems (ex. Microbiome). This book concludes with a section that explores the use of primary human cells in evaluating relevance to the clinic. 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.

Vital and authoritative, Type 2 Immunity: Methods and Protocols aims to provide a broad network of methods that can be used to develop a hypothesis and investigate its potential from bench to beside. 

1. A Fungal Protease Model To Interrogate Allergic Lung Immunity

J. Morgan Knight, Evan Li, Hui-Ying Tung, Cameron Landers, Jake Wheeler, Farrah Kheradmand, and David B Corry

Fulton, A., Babayan, S. A., and Taylor, M. D.

3. Production of Hymenolepis diminuta in the Laboratory: An Old Research Tool With New Clinical Applications

Min Zhang, Amanda J. Mathew, and William Parker

4. A Mouse Model of Peanut Allergy Induced By Sensitization through the Gastrointestinal Tract

Kelly Orgel and Michael Kulis

5. Induction and Characterization of the Allergic Eye Disease Mouse Model

6. Isolation and Purification of Epithelial and Endothelial Cells from Mouse Lung

Hideki Nakano, Keiko Nakano, and Donald N. Cook

7. In vitro and In vivo IgE/Antigen Mediated Mast Cell Activation

8. The use of Human and Mouse Mast Cell and Basophil Cultures to Assess

type-2 Inflammation

Heather L. Caslin, Marcela T. Taruselli, Anuya Paranjape, Kasalina Kiwanuka, Tamara Haque, Alena P. Chumanevich, Carole A. Oskeritzian, and John J. Ryan

9. Isolation and Identification of Group 2 Innate Lymphoid Cells in Settings of Type 2 Inflammation

Jesse Charles Nussbaum and Jorge Felipe Ortiz-Carpena

10. Determination of the Fate and Function of Innate Lymphoid Cells Following Adoptive Transfer of Innate Lymphoid Cell Precursors

Timothy E. O’Sullivan and Joseph C. Sun

11. Characterization of Thymic Development of Natural Killer T Cell Subsets by Multiparameter Flow Cytometry

Kathryn D. Tuttle and Laurent Gapin

Wanjiang Zeng, Rebecca L. O'Brien, Willi K. Born, and Yafei Huang

13. The Identification of Allergen-derived T cell Epitopes

Véronique Schulten and Alessandro Sette

14. Generation of Allergen-specific Tetramers for a Murine Model of Airway Inflammation

James J. Moon and Marion Pepper

16. Using Cytokine Reporter Mice to Visualize Type-2 immunity in vivo

Mark Dell’Aringa and R. Lee Reinhardt

17. Live Imaging of IL-4 expressing T Follicular Helper Cells in Explanted Lymph Nodes

Mark Dell’Aringa, R. Lee Reinhardt,Rachel S. Friedman, and Jordan Jacobelli

18. Imaging Precision Cut Lung Slices to Visualize Leukocyte Localization and Trafficking

Miranda R. Lyons-Cohen, Hideki Nakano, Seddon Y. Thomas, and Donald N. Cook

19. Study of IgE-producing B cells using the Verigem Fluorescent Reporter Mouse

Zhiyong Yang, James B. Jung, and Christopher D. C. Allen

20. Chromatin Preparation from Murine Eosinophils for Genome-wide Analyses

Carine Bouffi, Artem Barski, and Patricia C. Fulkerson

21. A Sensitive and Integrated Approach to Profile Messenger RNA from Samples with Low Cell Numbers

Sandy Lisette Rosales, Shu Liang, Isaac Engel, Benjamin Joachim Schmiedel, Mitchell

Kronenberg, Pandurangan Vijayanand, and Grégory Seumois

22. An Integrated and Semi-automated Micro-scaled Approach to Profile Cis-Regulatory Elements by Histone Modification ChIP-Seq for Large-scale Epigenetic Studies

23. Library preparation for ATAC-sequencing of Mouse CD4+ T Cells Isolated from the Lung and Lymph Nodes after Helminth Infection

Laura D. Harmacek, Preeyam Patel, Rachel Woolaver, R. Lee Reinhardt, and Brian P. O’Connor

24. Identification of Functionally-relevant microRNAs in the Regulation of Allergic Inflammation

Marlys S. Fassett, Heather H. Pua, Laura J. Simpson, David F. Steiner, and K. Mark Ansel

Charles B. Smarr and Stephen D. Miller

26. Assessing the Mouse Intestinal Microbiota in Settings of type-2 Immune Responses

27. The use of CRISPR-Cas9 Technology to Reveal Important Aspects of Human Airway Biology

Azzeddine Dakhama and Hong Wei Chu

28. A Consistent Method to Identify and Isolate Mononuclear Phagocytes from Human Lung and Lymph Nodes

Sophie L. Gibbingsand Claudia V. Jakubzick

29. Organoid Cultures for Assessing Intestinal Epithelial Differentiation and Function in Response to Type-2 Inflammation

Bailey Zwarycz, Adam D Gracz, and Scott T Magness

30. Utilization of Air-liquid Interface Cultures as an in vitro Model to Assess Primary Airway Epithelial Cell Responses to the Type 2 Cytokine Interleukin 13

Jamie L. Everman, Cydney Rios, and Max A. Seibold

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts