Main description:

This volume provides all the essential protocols that are currently used to study the immune synapse. Chapters in The Immune Synapse: Methods and Protocols cover methods for the study of the dynamics of immune synapse assembly, traffic at the immune synapse, new high resolution imaging, biophysical and computational methods for the study of the immune synapse, effector immune synapses, B cell, NK and mast cell immune synapses, and immune interactions in vivo. 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, The Immune Synapse: Methods and Protocols will be of interest to immunologists and, at a more general level, to cell biologists, biophysicists and computational biologists.

Contents:

1. The Immune Synapse: Past, Present, and Future

Michael L Dustin and Cosima T Baldari

2. Analyzing Actin Dynamics of the Immunological Synapse

Katarzyna I Jankowska and Janis K Burkhardt

3. Analysis of Microtubules and Microtubule-Organizing Centre at the Immune Synapse

Noelia Blas-Rus, Eugenio Bustos-Moran, Francisco Sanchez-Madrid, and Noa B Martin-Cofreces

4. Analyzing the Dynamics of Signaling Microclusters

Akiko Hashimoto-Tane, Tadashi Yokosuka, and Takashi Saito

5. Reconstitution of TCR Signaling using Supported Lipid Bilayers

Xiaolei Su, Jonathon A Ditlev, Michael K Rosen, and Ronald D Vale

6. Plasma Membrane Sheets for Studies of B Cell Antigen Internalization From Immune Synapses

Carla R. Nowosad and Pavel Tolar

7. Studying The Dynamics Of TCR Internalization At The Immune Synapse

Enrique Calleja, Balbino Alarcon, and Clara L Oeste

8. T Cell Receptor Activation of NF- b in Effector T Cells: Visualizing Signaling Events Within and Beyond the Cytoplasmic Domain of the Immunological Synapse

Maria Traver, Suman Paul, and Brian Schaefer

9. Imaging Vesicular Traffic at the Immune Synapse

Jerome Bouchet, Iratxe Del Rio-Iniguez, and Andres Alcover

10. Analysis of TCR Recycling at the Immune Synapse

Laura Patrussi and Cosima T Baldari

11. Simultaneous Membrane Capacitance Measurements and TIRF Microscopy to Study Granule Trafficking at Immune Synapses

Marwa Sleiman, David R. Stevens, and Jens Rettig

12. Mathematical Modeling of Synaptic Patterns

Anastasios Siokis, Philippe Robert, and Michael Meyer-Hermann

13. Super-Resolution Analysis of TCR-Dependent Signaling: Single Molecule Localization Microscopy

Valerie A Barr, Jason Yi, and Lawrence E Samelson

14. Foerster Resonance Energy Transfer to Study TCR-pMHC Interactions in the Immunological Synapse

Gerhard Schutza and Johannes B Huppa

15. Two-Dimensional Analysis of Cross-Junctional Molecular Interaction by Force Probes

Lining Ju, Yunfeng Chen, Muaz Nik Rushdi, Wei Chen, and Cheng Zhu

16. Studying Dynamic Plasma Membrane Binding of TCR-CD3 Chains during Immunological Synapse Formation using Donor-Quenching FRET and FLIM-FRET

Etienne Gagnon, Audrey Connolly, Jessica Dobbins, and Kai W Wucherpfennig

17. Revealing the Role of Microscale Architecture in Immune Synapse Function through Surface Micropatterning

Joung-Hyun Lee and Lance C Kam

18. Spatial Control of Biological Ligands on Surfaces

Haogang Cai, David Depoil, James Muller, Michael P.Sheetz, Michael L Dustin, and

Shalom J Wind

19. Probing Synaptic Biomechanics using Micropillar Arrays

Weiyang Jin, Charles T Black, Lance C Kam, and Morgan Huse

20. Microchannels for Study of Immunological Synapses and Kinapses

Helene D Moreau, Philippe Bousso, and Ana-Maria Lennon-Dumenil

21. Purification of LAT Containing Membranes from Resting and Activated T Lymphocytes

Claire Hivroz, Paola Larghi, Mabel Jouve, and Laurence Ardouin

22. Quantitative Phosphoproteomic Analysis of T-Cell Receptor Signaling

Nagib Ahsan and Arthur R Salomon

23. Imaging Asymmetric T Cell Division

Mirren Charnley and Sarah M Russell

24. Ultrastructure Of Immune Synapses

Jaime Llodra

25. Systems Imaging of the Immune Synapse

Rachel Ambler, Xiangtao Ruan, Robert F Murphy, and Christoph Wulfing

26. Comprehensive Analysis of Immunological Synapse Phenotypes using Supported Lipid Bilayers

Salvatore Valvo, Viveka Mayya, Jehan Afrose, Elena Seraia, Daniel Ebner, and Michael L. Dustin

27. Studying Immunoreceptor Signaling in Human T Cells using Electroporation of In Vitro Transcribed mRNA

Omkar Kawalekar, Carl H June, and Michael C. Milone

28. A Protein Expression Tool-Kit for Studying Signalling in T Cells

Ana Mafalda Santos, Jiandong Huo, Deborah Hatherley, Mami Chirifu, and Simon J Davis

29. Imaging The Effector CD8 Synapse

Gordon L Frazer, Yukako Asano, and Gillian M Griffiths

30.The Mast Cell Antibody-Dependent Degranulatory Synapse

Salvatore Valitutti, Regis Joulia, and Eric Espinosa

31. Measurement of Lytic Granule Convergence after Formation of an NK Cell Immunological Synapse

Hsiang-Ting Hsu, Alexandre F Carisey, and Jordan S Orange

32. Studying the T Cell-Astrocyte Immune Synapse

George P Cribaro, Elena Saavedra-Lopez, Paola V Casanova, Laura Rodriguez, and Carlos Barcia

33. Aberrant Immunological Synapses Driven by Leukemic Antigen Presenting Cells

Fabienne Mcclanahan and John G Gribben

34. Studying the Immune Synapse in HIV-1 Infection Iratxe Del Rio-Iniguez, Jerome Bouchet, and Andres Alcover

35. In Vivo Imaging of T Cell Immunological Synapses and Kinapses in Lymph Nodes

Helene D. Moreau and Philippe Bousso

36. Studying Dendritic Cell-T Cell Interactions Under In Vivo Conditions

Nicholas Van Panhuys