Immune Checkpoint Blockade: Methods and Protocols

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 Coa?a

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 Wickstr?m and Tanja L?vgren

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

Nataly Cruz Rodr?guez, Johanna Lineros, Carol Stefany Rodr?guez, Lina Mar?a Mart?nez, and Josefa Antonia Rodr?guez

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 Wickstr?m, Rolf Kiessling, and Yago Pico de Coa?a

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 Coa?a

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 Coa?a

Part III: Animal Models

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

Krist