Beschreibung
Cytokines are pleiotropic regulatory proteins involved in essentially all biological processes and associated with a wide variety of diseases, including inflammatory disorders as well as many types of cancer and leukemia. Knowledge about the quantitative and qualitative nature of cytokine production is critical in the understanding of normal and pathological processes. The cytokine detection in biological and clinical samples faces many challenges including their low abundance, the need to distinguish between active and latent cytokine forms, and the need to measure multiple cytokines in a single assay. This volume will provide a comprehensive collection of classic and cutting-edge methodologies that are currently used to analyze and quantify cytokines and their biological activities in complex biological and clinical samples. The chapters are divided into four main categories. The first group focuses on the immunodetection of released cytokines in tissue culture supernatants, plasma, serum and whole blood samples by immunoassays. These immunoassays measure the total concentrations of released cytokines regardless of their biological activity and include ELISA, flow cytometry, ELISPOT and the antibody-based proximity ligation. The second group will focus on the analysis of biologically active cytokines by bioassays using neutralizing antibodies, chemotaxis assay, cytokine-induced cell degranulation assay, cell proliferation and differentiation, cytokine-induced cytokine production and the radioreceptor cytokine assay. The third group focuses on the analysis of intracellular cytokines by flow cytometry, western blotting and fluorescence and confocal microscopy. In addition, this category includes protocols for quantitative analysis of cytokine gene expression by real time RT-PCR and analysis of the cytokine promoter occupancy by chromatin immunoprecipitation. The fourth group focuses on the recently developed multiplex arrays that can measure multiple cytokines in the same sample at the same time. This group includes quantification of multiple cytokines using cytometric bead arrays, ELISPOT assays, proteomics cytokine evaluation, multiplexed proximity ligation assays for high-throughput cytokine analysis and finally, cytokine gene expression analysis by gene arrays. The protocols will be written by experienced basic and clinical researchers with hands-on knowledge of the described protocols. By covering a broad variety of methods used in cytokine detection and analysis, this book will be of interest not only to biochemists, molecular biologists and immunologists but also to physician-scientists working in the field of cytokine research.
Autorenporträt
InhaltsangabePart I: Detection of Released Cytokines by Immunoassays - The Challenge of Measuring Elusive Immune Markers by Enzyme Linked Immuno-Spot (ELISPOT) Technique Maria Faresjö - Analysis of the Released Nuclear Cytokine HMGB1 in Human Serum Haichao Wang, Lin Zhao, Jianhua Li, Shu Zhu, and Meggie Yeung - aELISA Based Assay for IP-10 Detection from Filter Paper Samples Camilla Heldbjerg Drabe, Thomas Blauenfeldt, and Morten Ruhwald - Enhanced ELISA Based on Carboxymethylated Dextran Coatings Benoît Liberelle, Charles Fortier, and Gregory De Crescenzo - A Novel Three-Dimensional Biosensor Based on Aluminum Oxide: Application for Early Stage Detection of Human Interleukin-10 Michael Lee, Abdoullatif Baraket, Nadia Zine, Miguel Zabala, Francesca Campabadal, Raffaele Caruso, Maria Giovanna Trivella, Nicole Jaffrezic-Renault, and Abdelhamid Errachid 6. Quantification of Multiple Cytokines and Chemokines Using Cytometric Bead ArraysGemma Moncunill, Joseph J. Campo, and Carlota Dobaño 7. A Useful Guide for Analysis of Immune Markers by Fluorochrome (Luminex) Technique Maria FaresjöPart II: Cytokine Bioassays 8. Control of Pro-inflammatory Cytokine Release from Human Monocytes with the Use of an Interleukin-10 Monoclonal AntibodyHardik Patel and Dennis Davidson 9. Cytokine-Induced Neutrophil Chemotaxis Assay Dennis Davidson and Hardik Patel 10. An In Vitro One-Dimensional Assay to Study Growth Factor-Regulated Tumor Cell-Macrophage Interaction Ved P. Sharma, Brian T. Beaty, Dianne Cox, John S. Condeelis, and Robert J. Eddy 11. Chemotactic Responses by Macrophages to a Directional Source of a Cytokine Delivered by a Micropipette Michael Cammer and Dianne Cox 12. Assessment of Phagocytic Activity of Cultured Macrophages Using Fluorescence Microscopy and Flow Cytometry Lokesh Sharma, Wenjun Wu, Sanjay L. Dholakiya, Samir Gorasiya, Jiao Wu, Ravikumar Sitapara, Vivek Patel, Mao Wang, Michelle Zur, Shloka Reddy, Nathan Siegelaub, Katrina Bamba, Frank A. Barile, and Lin L. Mantell 13. Assessment of Cytokine-Modulated Proteasome Activity Christopher J. Kirk, Saul R. Powell, and Edmund J. Miller 14. Evaluation of the Adverse Effect of Low Concentration of Cadmium on Interleukin-4 Induced Class Switch Recombination in Burkett's Lymphoma Raji Cell Line Vladimir Poltoratsky 15. A Mix-and-Measure Assay for Determining the Activation Status of Endogenous Cdc42 in Cytokine-Stimulated Macrophage Cell Lysates Veronika Miskolci, Désirée Spiering, Dianne Cox, and Louis Hodgson 16. Analysis of the Cell Surface Expression of Cytokine Receptors Using the Surface Protein Biotinylation Method Mahmud Arif Pavel, Clarissa Lam, Parul Kashyap, Zahra Salehi-Najafabadi, Gurpreet Singh, and Yong Yu 17. Detection of CXCR2 Cytokine Receptor Surface Expression Using ImmunofluorescenceClarissa Lam, Mahmud Arif Pavel, Parul Kashyap, Zahra Salehi-Najafabadi, Victoria Valentino, and Yong Yu 18. Detecting Tie2, an Endothelial Growth Factor Receptor, by Using Immunohistochemistry in Mouse Lungs Prajna P. Guha, Sascha A. David, and Chandra C. Ghosh 19. Use of shRNA to Stably Suppress Chemokine Receptor Expression and Function in Human Cancer Cell Lines Nicole Salazar, Daniel Muñoz, James Hoy, and Bal L. Lokeshwar Part III. Analysis of Cytokine Intracellular protein levels and Gene Expression 20. Intracellular Staining and Detection of Cytokines by Fluorescence-Activated Flow Cytometry Giulia Freer 21. Cytokine Detection by Flow Cytometry Jian-Ge Qiu, Xiao-Long Mei, Zhe-Sheng Chen, and Zhi Shi 22. Analysis of IL-17 Production by Flow Cytometry and ELISPOT Assays Ling Zhao, Yu
