T cell activation entails a complex series of signal transduction events that begin with ligation of the T cell receptor by a cognate antigen, displayed on the surface of an antigen presenting cell. During T cell signaling proteins in the cell surface partition from one another into large, stereotyped molecular pattern that is known as the immunological synapse. This thesis attempts to understand the mechanisms by which this segregation occurs, and uses a number of fluorescence imaging techniques---in particular, single molecule microscopy---to this end. Contrary to reports that suggest a role for lipid raft domains in patterning the synapse, we find that protein-protein interactions, and not putative lipid raft associations, are a strong driving force in synapse formation. We also develop a method for reconstituting synapse formation in an immortalized T cell line that allows us to perform single molecule imaging on a fluid substrate of a defined composition. A common finding is that both passive mechanisms, involving diffusional trapping and exclusion, as well as active mechanisms, involving actin-driven transport, can act in concert to shape the immunological synapse.

This new collection features the most up-to-date essential protocols that are currently being used to study the immune synapse. Beginning with methods for making biophysical measurements, the volume continues by covering the cell biology of synapses, methods for advanced substrate engineering, mechanobiology topics, new technologies to describe and manipulate synaptic components, as well as methods related to sites of action and immunotherapy. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and fully updated, The Immune Synapse: Methods and Protocols, Second Edition serves as an ideal practical guide for researchers working in this dynamic field. Chapters 5, 11,18, 27, 30, and 32 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

A journey into the sub-microscopic world of molecular machines. Readers are first introduced to the types of molecules built by cells: proteins, nucleic acids, lipids, and polysaccharides. Then, in a series of distinctive illustrations, the reader is guided through the interior world of cells, exploring the ways in which molecules work in concert to perform the processes of living. Finally, the author shows us how vitamins, viruses, poisons, and drugs each have their effects on the molecules in our bodies. David Goodsell, author and illustrator, has prepared a fascinating introduction to biochemistry for the non-specialist. His book combines a lucid text with an abundance of drawings and computer graphics that present the world of cells and their components in a truly unique way.

Calcium Entry Channels in Non-Excitable Cells focuses on methods of investigating the structure and function of non-voltage gated calcium channels. Each chapter presents important discoveries in calcium entry pathways, specifically dealing with the molecular identification of store-operated calcium channels which were reviewed by earlier volumes in the Methods in Signal Transduction series. Crystallographic and pharmacological approaches to the study of calcium channels of epithelial cells are also discussed. Calcium ion is a messenger in most cell types. Whereas voltage gated calcium channels have been studied extensively, the non-voltage gated calcium entry channel genes have only been identified relatively recently. The book will fill this important niche.

This volume details our current understanding of the architecture and signaling capabilities of the B cell antigen receptor (BCR) in health and disease. The first chapters review new insights into the assembly of BCR components and their organization on the cell surface. Subsequent contributions focus on the molecular interactions that connect the BCR with major intracellular signaling pathways such as Ca2+ mobilization, membrane phospholipid metabolism, nuclear translocation of NF-kB or the activation of Bruton’s Tyrosine Kinase and MAP kinases. These elements orchestrate cytoplasmic and nuclear responses as well as cytoskeleton dynamics for antigen internalization. Furthermore, a key mechanism of how B cells remember their cognate antigen is discussed in detail. Altogether, the discoveries presented provide a better understanding of B cell biology and help to explain some B cell-mediated pathogenicities, like autoimmune phenomena or the formation of B cell tumors, while also paving the way for eventually combating these diseases.

"Taken together, the body of information contained in this book provides readers with a bird’s-eye view of different aspects of exciting work at the convergence of disciplines that will ultimately lead to a future where we understand how immunity is regulated, and how we can harness this knowledge toward practical ends that reduce human suffering. I commend the editors for putting this volume together." –Arup K. Chakraborty, Robert T. Haslam Professor of Chemical Engineering, and Professor of Physics, Chemistry, and Biological Engineering, Massachusetts Institute of Technology, Cambridge, USA New experimental techniques in immunology have produced large and complex data sets that require quantitative modeling for analysis. This book provides a complete overview of computational immunology, from basic concepts to mathematical modeling at the single molecule, cellular, organism, and population levels. It showcases modern mechanistic models and their use in making predictions, designing experiments, and elucidating underlying biochemical processes. It begins with an introduction to data analysis, approximations, and assumptions used in model building. Core chapters address models and methods for studying immune responses, with fundamental concepts clearly defined. Readers from immunology, quantitative biology, and applied physics will benefit from the following: Fundamental principles of computational immunology and modern quantitative methods for studying immune response at the single molecule, cellular, organism, and population levels. An overview of basic concepts in modeling and data analysis. Coverage of topics where mechanistic modeling has contributed substantially to current understanding. Discussion of genetic diversity of the immune system, cell signaling in the immune system, immune response at the cell population scale, and ecology of host-pathogen interactions.