The Latest on Immune Tolerance

The campus here at CSHL was abuzz last week with the 78^th annual Cold Spring Harbor Symposium, where scientists from around the world assembled to discuss immunity and tolerance – also the topic of our new book Immune Tolerance.

“Immunological tolerance is a phenomenon that has fascinated immunologists for decades,” explained Diane Mathis in an interview at the meeting last week. “The immune system was set up to fight a whole diversity of different kinds of challenges. But sometimes [immune cells] are able to see a component of the body itself. There must be ways to deal with this issue . . . so they can’t attack the body’s own cells.”

In the book, which Diane coedited with Alexander Rudensky, the contributors discuss our current understanding of the mechanisms involved in establishing and maintaining immune tolerance, ensuring that the immune system responds to foreign molecules and not to self-molecules. The contributors also describe factors that underlie the breakdown of tolerance, leading to autoimmune diseases such as rheumatoid arthritis, lupus, and multiple sclerosis, and strategies to restore tolerance in clinical settings. The book will be a valuable reference for all immunologists and clinicians wishing to understand or develop treatments for autoimmune diseases.

For more information on the book, click here.

 

Understanding Cell Death

Means to an End: Apoptosis and Other Cell Death Mechanisms is the new book from Douglas Green, a clear and comprehensive view of apoptosis and other death mechanisms. He examines the enzymes that perform the execution (caspases) and the molecular machinery that links their activation to signals that cause cell death, emphasizing the importance of BCL-2 proteins and cytochrome c released from mitochondria. Green also outlines the roles of cell death in embryogenesis, neuronal selection, and the development of self-tolerance in the immune system, explains how cell death defends the body against cancer, and traces the evolutionary origins of the apoptosis machinery back over a billion years.

There’s also an online companion resource that’s rapidly growing. Be sure not to miss Cell Death: The Movie.

 

Drosophila Immunohistochemistry

Cold Spring Harbor Laboratory Press’ new Drosophila Neurobiology laboratory manual covers the three main approaches taught in the CSHL course: studying neural development, recording and imaging the nervous system, and studying behavior. The featured electrophysiology paper is part of the recording/imaging section, while the second featured article in the July issue of Cold Spring Harbor Protocols comes from a neural development chapter.

The larval Drosophila brain has been a valuable model for investigating the role of stem cells in development. These neural stem cells, called “neuroblasts,” have provided insight into the role of cell polarity in influencing cell fate. Identifying neuroblasts and their progeny requires a method capable of recognizing cell polarity and cell fate markers. Immunofluorescent Staining of Drosophila Larval Brain Tissue, provided by Cheng-Yu Lee and colleagues, describes procedures for the collection and processing of Drosophila larval brains for analysis of these markers. Neuroblasts are identified via immunolocalization, the use of labeled antibodies that specifically bind the marker proteins of interest. As one of our featured articles, it is freely available to subscribers and non-subscribers alike.

 

Monitoring cellular immune response

The enzyme-linked immunospot (ELISPOT) assay is considered by many to be the gold standard for monitoring cellular immune responses. The method is highly sensitive, quantitative, easy to use and amenable to high throughput screening. Until recently, the ELISPOT assay has been limited to the characterization of only one single effector molecule. Since the maintenance of both IFN-gamma and IL-2 by pathogen-specific T cells has been linked to a more favorable clinical outcome in human immunodeficiency virus (HIV) and Leishmania infections, an ELISPOT assay able to characterize both these effector molecules would be helpful for monitoring immune responses to certain infectious agents. Nicole Bernard and colleagues from the McGill University Health Centre present a protocol for Dual-Color ELISPOT Assay for the Simultaneous Detection of IL-2 and/or IFN-gamma Secreting T Cells in the January issue of Cold Spring Harbor Protocols. As interest in multifunctional T-cell monitoring in human diseases grows, this method is likely to be extensively used. The protocol is one of January’s featured articles, and is freely available to subscribers and non-subscribers alike.

 

Lymphoctye differentiation from stem cells

February’s issue of CSH Protocols has just posted, and one of our featured articles this month is The OP9-DL1 System: Generation of T-Lymphocytes from Embryonic or Hematopoietic Stem Cells In Vitro, from Juan Carlos Zúñiga-Pflücker’s laboratory at the University of Toronto. By using OP9-DL1 cells as a support system, researchers can study the differentiation of embryonic stem cells into mature components of the immune system. The article contains a series of protocols describing the establishment, maintenance, and storage of OP9 and OP9-DL1 cells; the co-culture of these cells with embryonic stem cells or hematopoietic stem cells from fetal liver or bone marrow; and the in vitro differentiation of the stem cells into lymphocytes. The OP9-DL1 system has been useful in addressing questions about the cellular and molecular regulation of T-lymphocyte lineage commitment, pre-T cell receptor signaling (Beta-selection), functional characteristics of progenitor T cells, and maturation of functional CD8 T cells.

Like all of our featured articles, this set of protocols is freely available to subscribers and non-subscribers.

 

New free mini-lab manual available at ASCB Meeting

Cold Spring Harbor Laboratory Press has long been known for the creation of influential laboratory manuals like Molecular Cloning (aka “Maniatis”). CSH Protocols was created in order to move our manual publishing program into the online world. One of the interesting opportunities this creates is the ability to, in a sense, reverse engineer things. Instead of commissioning a manual, publishing it as a book and adding the protocols to the journal’s collection, we’re also interested in creating new print manuals based on the journal’s content.

New printing technologies have made it easier to produce high quality books and magazines. It’s now becoming feasible for us to re-mix and re-combine material from a variety of sources into a small, focused volume on one particular subject. Our first attempt is a collection of articles detailing methods for Immunohistochemistry, chosen because it is in such widespread use in so many fields of biological research. While not a comprehensive treatment of the topic, the collection covers the basic methods and includes specialized protocols for common laboratory organisms (yeast, plants, C. elegans, Drosophila, zebrafish, Xenopus and mouse). Our generous sponsors (Santa Cruz Biotechnology, Sigma-Aldrich, AbD Serotec and R&D Systems) have enabled us to distribute this experiment to you free of charge.

For now, this mini-manual is only available the CSHL Press booth at major meetings, and in Grace Auditorium at Cold Spring Harbor Laboratory meetings. Please come by our booth at the ASCB Meeting (Booth #311) this week and grab a copy for your lab.

 

Immunohistochemistry coverage expanded

Immunohistochemistry (the localization of proteins in a tissue by binding antibodies to specific antigens) is a technique where one protocol definitely does not fit all. Each model organism seems to have its own quirks, whether it be in the fixative used, the methods needed for antibody penetration, issues with autofluorescence or even just figuring out which cross-species antibodies work in a given system. To that end, we’ve been working on expanding our coverage of immunohistological protocols. The February issue of CSH Protocols brings methods for plant sections, using both avidin-biotin and alk-phos, as well as a method for whole-mount immunocytochemistry in Xenopus embryos from John Wallingford’s lab at the University of Texas (they provided the lovely cover image for this month).

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September Song

September’s issue of CSH Protocols is upon us (my, that was a quick summer–Kurt Weill sure knew what he was talking about). We’re featuring two protocols this month, both available freely to non-subscribers (as are all of our sample protocols). The first is for generating mouse models for squamous cell carcinoma and the second is for the Oligonucleotide Ligation Assay (OLA), used for finding Single-Nucleotide Polymorphisms (SNP’s).