Main description:

This volume provides key methods and protocols from laboratories engaged in germinal centers (GC) research with the expectation of stimulating further research, and to aid scientists in the study of GC biology and pathology. 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 practical, Germinal Centers: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

1. Analysis of the Germinal Center Reaction in Tissue Sections

David Dominguez-Sola and Giorgio Cattoretti

2. Detecting Gene Expression in Lymphoid Microenvironments by Laser Microdissection and Quantitative RT-PCR

Yang Zhang, Laura Garcia-Ibanez, Geoffrey Brown, and Kai-Michael Toellner

3. Tracking Plasma Cell Differentiation in Living Mice with 2-Photon Microscopy

Carolin Ulbricht, Randall L. Lindquist, Laura Tech, and Anja E. Hauser

4. Microanatomical Labeling of Germinal Center Structures for Flow Cytometry Using Photoactivation

Johanne T. Jacobsen and Gabriel D. Victora

5. Fate Mapping and Transcript Profiling of Germinal Center Cells by Two-photon Photoconversion

Imogen Moran and Tri Giang Phan

6. Intravital Microscopy of T-B Cell Interactions in Germinal Centers

Changming Shih and Hai Qi

7. Identifying Follicular Regulatory T cells by Confocal MicroscopyVanderleyden I, Linterman MA

8. Cytokine Expression by T follicular Helper Cells

Christoph Jandl, Claudia Loetsch, and Cecile King

9. Follicular Dendritic Cell Isolation and Loading of Immune Complexes

Balthasar A. Heesters, Cornelis E. Van der Poel, and Michael C. Carroll

10. Isolation and Characterization of Mouse and Human Follicular Dendritic Cells

Mohey Eldin M. El Shikh, Riham El Sayed, and Costantino Pitzalis

11. In-vitro-induced Germinal Center B Cell Culture System

Kei Haniuda, Takuya Nojima and Daisuke Kitamura

12. CRISPR/Cas9 Mediated In Vitro Mutagenesis in GC-like B Cells

Van Trung Chu, Robin Graf and Klaus Rajewsky

13. Germinal Center Formation with Retrovirally-transduced B Cells for Determining the Role of Specific Molecules In Vivo

Rinako Nakagawa

14. Characterization of the B Cell Transcriptome by RNA-Binding Proteins using iCLIP

Manuel D. Diaz-Munoz, Elisa Monzon-Casanova, and Martin Turner

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15. Flow-Cytometric Method Measuring B Cell Surface Immunoglobulin Avidity

Davide Angeletti, Gregory M. Frank, and Jonathan W. Yewdell

16. Somatic Hypermutation and Affinity Maturation Analysis Using the 4-hydroxy-3-nitrophenyl-acetyl (NP) System

Nicole Heise and Ulf Klein

17. Targeting Gene Function in Germinal Center B Cells: A Practical Approach

Valentina Petrocelli and Stefano Casola

18. Development of Mouse Model Systems of Germinal Centre Lymphomas

Eleni Kabrani and Sandrine Sander

19. The AID-Cre-ERT2 Model: A Tool for Monitoring B Cell Immune Responses and Generating Selective Hybridomas

Simon Le Gallou, Takuya Nojima, Daisuke Kitamura, Jean-Claude Weill, and Claude-Agnes Reynaud

20. Determining the Origin of Human Germinal Center B Cell-derived Malignancies

Marc Seifert and Ralf Kuppers

21. Tracking B-cell Repertoires and Clonal Histories in Normal and Malignant Lymphocytes

Nicola J. Weston-Bell, Graeme Cowan, and Surinder S. Sahota

22. How to Simulate a Germinal Centre

Philippe A. Robert, Ananya Rastogi, Sebastian Binder, and Michael Meyer-Hermann