Cytokine Bioassays

1. Front Matter

   Pages i-xiv

   PDF

2. Detection of Released Cytokines by Immunoassays

   1. Front Matter

      Pages 1-1

      PDF

   2. The Challenge of Measuring Elusive Immune Markers by Enzyme-Linked Immuno-Spot (ELISPOT) Technique

      • Maria Faresjö

      Pages 3-12

   3. Analysis of the Released Nuclear Cytokine HMGB1 in Human Serum

      • Haichao Wang, Lin Zhao, Jianhua Li, Shu Zhu, Maggie Yeung

      Pages 13-25

   4. ELISA-Based Assay for IP-10 Detection from Filter Paper Samples

      • Camilla Heldbjerg Drabe, Thomas Blauenfeldt, Morten Ruhwald

      Pages 27-37

   5. Enhanced ELISA Based on Carboxymethylated Dextran Coatings

      • Benoît Liberelle, Charles Fortier, Gregory De Crescenzo

      Pages 39-47

   6. 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 et al.

      Pages 49-64

   7. Quantification of Multiple Cytokines and Chemokines Using Cytometric Bead Arrays

      • Gemma Moncunill, Joseph J. Campo, Carlota Dobaño

      Pages 65-86

   8. A Useful Guide for Analysis of Immune Markers by Fluorochrome (Luminex) Technique

      • Maria Faresjö

      Pages 87-96

3. Cytokine Bioassays

   1. Front Matter

      Pages 97-97

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   2. Control of Pro-inflammatory Cytokine Release from Human Monocytes with the Use of an Interleukin-10 Monoclonal Antibody

      • Hardik Patel, Dennis Davidson

      Pages 99-106

   3. Cytokine-Induced Neutrophil Chemotaxis Assay

      • Dennis Davidson, Hardik Patel

      Pages 107-113

   4. 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, Robert J. Eddy

      Pages 115-123

   5. Chemotactic Responses by Macrophages to a Directional Source of a Cytokine Delivered by a Micropipette

      • Michael Cammer, Dianne Cox

      Pages 125-135

   6. 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 et al.

      Pages 137-145

   7. Assessment of Cytokine-Modulated Proteasome Activity

      • Christopher J. Kirk, Saul R. Powell, Edmund J. Miller

      Pages 147-162

   8. 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

      Pages 163-171

   9. 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, Louis Hodgson

      Pages 173-184

   10. 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, Yong Yu

       Pages 185-192

   11. Detection of CXCR2 Cytokine Receptor Surface Expression Using Immunofluorescence

       • Clarissa Lam, Mahmud Arif Pavel, Parul Kashyap, Zahra Salehi-Najafabadi, Victoria Valentino, Yong Yu

       Pages 193-200

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 multiplecytokines 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.

• Cytokines
• RT-PCR
• analysis of intracellular cytokines by flow cytometry
• biologically active cytokines
• clinical samples
• cytokine detection
• immunodetection of released cytokines
• immunofluorescence confocal microscopy
• immunohistochemistry
• quantitative analysis of cytokine gene expression
• western blotting

• Department of Biological Sciences, St. John's University, New York, USA

  Ivana Vancurova

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