Main description:

This detailed volume describes a series of techniques that are essential for evaluating the efficacy of new checkpoint blockade therapies as well as understanding the mechanisms behind the therapies that have already been approved. Beginning with a section on describing the tumor microenvironment and evaluating the immune system at a systemic level, the book continues by covering functional assays that provide answers to questions that may be raised after studying the immune system and its responses to immunotherapies, as well as the use of animal models in this research. Written for the highly successful Methods in Molecular Biology series, 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.
Authoritative and practical, Immune Checkpoint Blockade: Methods and Protocols serves to aid researchers in furthering our understanding of checkpoint blockage as well as the study of tumor immunology and the development of new immunotherapies.

Contents:

Part I: Generating Hypothesis

1. Double Immunohistochemistry and Digital Image Analysis

Pablo Moreno-Ruiz, Lina Wik Leiss, Artur Mezheyeuski, and Monika Ehnman

2. Multispectral Fluorescence Imaging Allows for Distinctive Topographic Assessment and Sub-Classification of Tumor Infiltrating and Surrounding Immune Cells

Claudia Wickenhauser, Daniel Bethmann, Zipei Feng, Shawn M. Jensen, Carmen Ballesteros-Merino, Chiara Massa, Andre Steven, Marcus Bauer, Peter Kaatzsch, Nikolaos Pazaitis, Georgiana Toma, Carlo B. Bifulco, Bernard A. Fox, and Barbara Seliger

3. Systems Level Immune Monitoring by Mass Cytometry

Tadepally Lakshmikanth and Petter Brodin

4. Immune Monitoring of Cancer Patients by Multi-Color Flow Cytometry

Neo Shiyong, Aine O'Reilly, and Yago Pico de Coana

5. High-Throughput, Fast and Sensitive Immunopeptidomics Sample Processing for Mass Spectrometry

Fabio Marino, Chloe Chong, Justine Michaux, and Michal Bassani-Sternberg

6. Identification of Immune Modulatory miRNAs by miRNA Enrichment via RNA Affinity Purification

Uta Sandy Tretbar, Michael Friedrich, Maria-Filothei Lazaridou, and Barbara Seliger

Part II: Functional Assays

7. Expansion of Tumor-Infiltrating Lymphocytes from Melanoma Tumors

Stina Wickstroem and Tanja Loevgren

8. Establishment of Bi-Dimensional (2D) and Three-Dimensional (3D) Melanoma Primary Cultures as a Tool for In Vitro Drug Resistance Studies

Nataly Cruz Rodriguez, Johanna Lineros, Carol Stefany Rodriguez, Lina Maria Martinez, and Josefa Antonia Rodriguez

9. Assessment of Anti-Tumor T Cell Responses by Flow Cytometry after Co-Culture of Tumor Cells with Autologous Tumor-Infiltrating Lymphocytes

Jeroen Melief, Stina Wickstroem, Rolf Kiessling, and Yago Pico de Coana

10. Expansion and Determination of Antigen-Reactive T Cells by Flow Cytometry

Alexander Martens, Graham Pawelec, and Christopher Shipp

11. Assessing Canonical and Adaptive Natural Killer Cell Function in Suppression Assays In Vitro

Dhifaf Sarhan and Jeffrey S. Miller

12. Evaluating Antibody-Dependent Cell-Mediated Cytotoxicity by Chromium Release Assay

Irene van der Haar Avila, Patricia Marmol, Rolf Kiessling, and Yago Pico de Coana

13. Evaluating Antibody-Dependent Cell-Mediated Cytotoxicity by Flow Cytometry

Irene van der Haar Avila, Patricia Marmol, Jeannette Cany, Rolf Kiessling, and Yago Pico de Coana

Part III: Animal Models

14. Evaluation of Breast Cancer and Melanoma Metastasis in Syngeneic Mouse Models

Kristina Witt and Andreas Lundqvist

15. Establishment of Melanoma Tumor Xenograft Using Single Cell Line Suspension and Co-Injection of Patient-Derived T Cells in Immune Deficient NSG Mice

Frida Bjoerk Gunnarsdottir, Rolf Kiessling, and Yago Pico de Coana

16. Analysis of Tissue Resident Immune Cells from Mouse Skin and Lungs by Flow Cytometry

Felipe Galvez-Cancino, Ernesto Lopez, and Alvaro Lladser