Main description:

This book provides a comprehensive collection of classic and cutting-edge methodologies as well as bioinformatics and genome-editing approaches that are used to quantify immune mediators and analyze their function and biological activity in cancer cells and tissues. Beginning with a section on the detection of immune mediators in samples, the volume continues with sections covering cytokine bioassays, the expression and regulation of immune mediators in cancer cells, and methods to navigate the enormous datasets created by modern DNA and RNA sequencing and proteomic technology. 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.
Authoritative and wide-ranging, Immune Mediators in Cancer: Methods and Protocols serves as a valuable resource for biochemists, molecular biologists, cancer biologists, and immunologists, as well as for physician-scientists working in the field of immunology and cancer research.

Contents:

Part I: Detection of Immune Mediators in Cancer Cells and Tissues

1. A Useful Guide for Analysis of Immune Mediators in Cancer by Fluorochrome (Luminex) Technique

Maria Faresjoe

2. Analysis of Circulating HMGB1 in Human Serum

Weiqiang Chen, Guoqiang Bao, Lin Zhao, and Haichao Wang

3. Analysis of IL-22 and Th22 Cells by Flow Cytometry in Systemic Lupus Erythematosus

Zhuang Ye, Ling Zhao, Qi Gao, Yanfang Jiang, Zhenyu Jiang, and Cong-Qiu Chu

4. Application of Immunohistochemistry in Basic and Clinical Studies

Aihua Li and Dong-Hua Yang

5. Interleukin-1 (IL-1) Immunohistochemistry Assay in Oral Squamous Cell Carcinoma

Takaaki Kamatani

6. Luminex xMAP Assay to Quantify Cytokines in Cancer Patient Serum

Helena Kupcova Skalnikova, Katerina Vodickova Kepkova, and Petr Vodicka

7. Detection of Cytokine Receptors Using Tyramide Signal Amplification for Immunofluorescence

Herui Wang, Ryan L. Pangilinan, and Yan Zhu

Part II: Cytokine Bioassays

8. Analysis of IFNg-Induced Migration of Ovarian Cancer Cells

Bijaya Gaire, Mohammad M. Uddin, Yue Zou, and Ivana Vancurova

9. Interleukin-8-Induced Invasion Assay in Triple Negative Breast Cancer Cells

Mohammad M. Uddin, Bijaya Gaire, Betsy Deza, and Ivana Vancurova

10. Interleukin-8 Induces Proliferation of Ovarian Cancer Cells in 3D Spheroids

Mohammad M. Uddin, Bijaya Gaire, and Ivana Vancurova

11. Detection of Ferroptosis by BODIPY (TM) 581/591 C11

Alejandra M. Martinez, Ahryun Kim, and Wan Seok Yang

12. Methods for Studying TNF -Induced Autophagy

Sheyda Najafi, Ehab M. Abo-Ali, and Vikas V. Dukhande

13. Isolation of Antibody Binders to MISIIR from a Phage Display Library by Sorting

Andy Qingan Yuan

14. Measuring Chimeric Antigen Receptor T Cells (CAR T Cells) Activation by Coupling Intracellular Cytokine Staining with Flow Cytometry

Chong Xu and Yibo Yin

15. Analysis of Interleukin-4-Induced Class Switch Recombination in Mouse Myeloma CH12F3-2 Cells

Wenjun Wu, Zhihui Xiao, Deon Buritis, and Vladimir Poltoratsky

16. Single-Stranded DNA Aptamers against TNF and Their Potential Applications

Shao Tao, Song Pingfang, Xiaowei Zhang, Lingshu Zhang, and Cong-Qiu Chu

17. Probing Metabolic Changes in IFN -Treated Ovarian Cancer Cells

Pritpal Kaur, Shreya Nagar, Madhura Bhagwat, Mohammad Uddin, Yan Zhu, and Ales Vancura

Part III: Expression and Regulation of Immune Mediators in Cancer Cells

18. Immunoblotting Analysis of Intracellular PD-L1 Levels in Interferon-g-Treated Ovarian Cancer Cells Stably Transfected with Bcl3 shRNA

Sveta Padmanabhan, Yue Zou, and Ivana Vancurova

19. Flow Cytometry Analysis of Surface PD-L1 Expression Induced by IFNg and Romidepsin in Ovarian Cancer Cells

Sveta Padmanabhan, Yue Zou, and Ivana Vancurova

20. Analysis of PD-L1 Transcriptional Regulation in Ovarian Cancer Cells by Chromatin Immunoprecipitation

Yue Zou, Sveta Padmanabhan, and Ivana Vancurova

21. Real-Time PCR Assay for the Analysis of Alternative Splicing of Immune Mediators in Cancer

Ruizhi Wang, Md. Faruk Hossain, Jovan Mirkovic, Samuel Sabzanov, and Matteo Ruggiu

22. Combined Single-Cell Measurement of Cytokine mRNA and Protein in Immune Cells

Julian J. Freen-van Heeren, Benoit P. Nicolet, and Monika C. Wolkers

23. Microscopic Methods for Analysis of Macrophage-Induced Tunneling Nanotubes

Kiersten P. Carter, Jeffrey E. Segall, and Dianne Cox

24. Optogenetics: Rho GTPases Activated by Light in Living Macrophages

Maren Hulsemann, Polina V. Verkhusha, Peng Guo, Veronika Miskolci, Dianne Cox, and Louis Hodgson

Part IV: Bioinformatics and Genome Editing Approaches

25. High-Throughput RNA interference Screen Targeting Synthetic-Lethal Gain-of-Function of Oncogenic Mutant TP53 in Triple Negative Breast Cancer

Susumu Rokudai

26. Discovering Transcription Factor Non-Coding RNA Targets Using ChIP-Seq Analysis

Vitalay Fomin and Carol Prives

27. Phylogenetic Analyses of Chemokine Receptors from Sequence Retrieval to Phylogenetic Trees

Juan C. Santos

28. Generation of IL17RB Knockout Cell Lines Using CRISPR/Cas9-Based Genome Editing

Olivia Hu, Alessandro Provvido, and Yan Zhu

29. Engineering Mutation Clones in Mammalian Cells with CRISPR/Cas9

Zijun Huo, Jian Tu, Dung-Fang Lee, and Ruiying Zhao