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. 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 Coaa 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 Wickstrm and Tanja Lvgren 8. Establishment of Bi-Dimensional (2D) and Three-Dimensional (3D) Melanoma Primary Cultures as a Tool for In Vitro Drug Resistance Studies Nataly Cruz Rodrguez, Johanna Lineros, Carol Stefany Rodrguez, Lina Mara Martnez, and Josefa Antonia Rodrguez 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 Wickstrm, Rolf Kiessling, and Yago Pico de Coaa 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 Coaa 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 Coaa 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 Fra Bjrk Gunnarsdttir, Rolf Kiessling, and Yago Pico de Coaa 16. Analysis of Tissue Resident Immune Cells from Mouse Skin and Lungs by Flow Cytometry Felipe Galvez-Cancino, Ernesto Lopez, and Alvaro Lladser
