This volume is a practical biochemical guide to the Enzyme-Linked Immunosorbent Assay (ELISA), used to detect a target substance in a liquid sample. The ELISA is an important and widely used diagnostic tool in medicine, animal health, botany and quality assurance processes in food and beverage production. An introductory chapter orients the reader on the basic structure and function of immunoglobulins and their fragments while subsequent chapters outline the methodology to generate monoclonal antibodies using hybridoma technology and the general methods used to purify antibodies. Multiple chapters demonstrate how to creatively use the properties of the antibody to identify, localize and quantify target analytes to answer questions and resolve problems. The reader will learn how to use a variety of immunoassay strategies, reporters and detection systems that will undoubtedly facilitate their efforts to gain answers to their own questions. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, ELISA: Methods and Protocols seeks to provide both professionals and novices with the technical information necessary for the reader to successfully use the immunoassay as part of the discovery process.

Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industrial applications as well as in the progression of established and emerging production strategies. This volume provides detailed coverage of the generation, optimization, characterization, production and applications of antibody. It provides the necessary theoretical background and description of methods for the expression of antibody in microbial and animal cell cultures and in transgenic animals and plants. There is a strong focus on those issues related to the production of intrabodies, bispecific antibody and antibody fragments and also to novel applications in cancer immunotherapy.

The book covers recent developments in research and practice of allergy and immunology. Special emphasis has been given to epidemiology and the relation of genetic and environmental factors in allergic diseases. Occupational aspects and the pathophysiology are additionally covered and an overview of the current pharmacotherapy and immunotherapy is provided.

Introductory Address.- The New Biology and Vaccine Research.- Keynote Presentation.- Mucosal Immunity To Vaccines: Current Concepts for Vaccine Development and Immune Response Analysis.- Session I: Oral Diseases and Host Immune Responses.- Prospects for Human Mucosal Vaccines.- Bacterial Diseases of the Oral Tissues.- Oral Virus Infections: The Potential for Gene Transfer in Treatment and Prevention.- Session II: Update on Vaccines and Vaccine Development.- Bacterial Mucosal Vaccines.- A General Overview of Viral Vaccine Development.- Session III: Vaccines and the Mucosal Immune System.- An Update on the "Jennerian" and Modified "Jennerian" Approach to Vaccination of Infants and Young Children Against Rota Virus Diarrhea.- Induction of Mucosal and Serum Immune Responses to a Specific Antigen of Peridontal Bacteria.- IgA1 Proteases and Host-Parasite Relationships in the Oral Cavity.- Transport of Iga Immune Complexes Across Epithelial Membranes: New Concepts n Mucosal Immunity.- Effect of Mucosal Microenvironment on Immune Response to Viruses.- Session IV: Optimizing Mucosal and Systemic Immune Responses.- Induction of T Helper Cells and Cytokines For Mucosal IgA Responses.- Cytokine Production and T Cell Receptor Expression by Salivary Gland T Cell and Intraepithelial T Lymphocytes for the Regulation of the IgA Response.- Immunological Adjuvants.- Session V: Delivery Systems and Immune Analysis.- M Cell-Mediated Antigen Transport and Monoclonal IgA Antibodies For MucosalImmune Protection.- A Mechanism of Passive Immunization with Monoclonal Antibodies to a 185,000Mr Streptococcal Antigen.- Delivery of Antigens by Recombinant Avirulent Salmonella Strains.- Use of Recombinant BCG as a Vaccine Delivery Vehicle.- Vaccinia Virus Recombinants as Potential Herpes Simplex Virus Vaccines.- Liposomes and Conjugate Vaccines for Antigen Delivery and Induction of Mucosal Immune Responses.- Peroral Immunization with a Cholera Toxin-Linked Bacterial Protein Antigen and Synthetic Peptide.- Peptomers as Vaccine Candidates.- Session VI: Target Antigen Selection and Vaccine Development.- Structural and Functional Studies of Herpes Simplex Virus Glycoprotein D.- Molecular, Immunological and Functional Characterization of the Major Surfae Adhesin of Streptococcus Mutans.- Reacitve Antigens of the Periodontopathic Bacterium Actinobacillus Actinmycetemcomitans.- Immunization with Fimbrial Protein and Peptide Protects Against Porphyromonas Gingivalis- Induced Periodontal Tissue Destruction.- Vaccine Development: Progression from Target Antigen to Product.- Session VII: Immunological Correlates of Protection.- Significance of Immune Responses to Oral Antigens in Dental Diseases.- Laboratory Correlates of Protection and Protective Immunity to Bordetella Pertussis.- Future Directions.- Challenges and Opportunities in Vaccine Research.- Summary and Recommendations for Future Research.- Speakers and Moderators.- Author Index.

The immune system is not bound by a single tissue but is instead bestowed with the challenge of warding off invading pathogens throughout the body. Constant surveillance of the body requires that the immune system be highly mobile and able to purge pathogens from all tissues. Because each tissue presents its own unique architecture and milieu, it is necessary for the immune system to be as malleable as it is dynamic. For example, how the immune system handles a pathogen in the lung can differ significantly from a pathogen encountered in the gut.

Understanding immune complexity in diverse tissue environments is a challenge for researchers. However, advances in imaging have greatly improved our ability to probe the immune system. From snap-shots in time to 4D movies, imaging systems have been used to generate stunning visualizations of immune cells in action throughout the body. These visualizations are not only aesthetically pleasing but they have yielded great advances in our understanding of immune function. This volume provides a synopsis of major insights in immunology revealed using imaging approaches. "Seeing is truly believing," and this volume was assembled to recognize past accomplishments and to provide visions of what the future holds in store in this exciting field.

Established for almost 30 years, Methods in Microbiology is the most prestigious series devoted to techniques and methodology in the field. Now totally revamped, revitalized, with a new format and expanded scope, Methods in Microbiology will continue to provide you with tried and tested, cutting-edge protocols to directly benefit your research.
Immunology of Infection, edited by two of the foremost figures in the field, presents the most appropriate, up-to-date techniques in the detail you require. The layout is structured for ease of reference, and the volume will be essential reading for all researchers working in microbiology, immunology, virology, mycology, and parasitology.
The new volume provides a carefully selected collection of immunological techniques for the microbiologist wishing to study host-pathogen relationships "in vivo" and "in vitro." This multi-authored book has succeeded in bringing together experts from various fields of molecular and cellular immunology who provide ready-to-use recipes for the "ex vivo" and "in vitro" analysis of anti-infective immunity.
Key Features
* Focuses on the methods most useful for the microbiologist interested in analysing host-pathogen relationships
* Ready-to-use, tried and tested recipes
* Lists of suppliers provided as appendices to each chapter
* Covers techniques useful for the analysis of human and murine cells
* Includes techniques for the prediction and determination of MHC ligands and T cell epitopes
* Describes the art and science of DNA vaccines
* Essential methods for measuring human cytokine responses
* Covers isolation and propagation of dendritic cells

The ?eld of cellular responses to DNA damage has attained widespread recognition and interest in recent years commensurate with its fundamental role in the ma- tenance of genomic stability. These responses, which are essential to preventing cellular death or malignant transformation, are organized into a sophisticated s- tem designated the "DNA damage response". This system operates in all living organisms to maintain genomic stability in the face of constant attacks on the DNA from a variety of endogenous by-products of normal metabolism, as well as exogenous agents such as radiation and toxic chemicals in the environment. The response repairs DNA damage via an intricate cellular signal transduction network that coordinates with various processes such as regulation of DNA replication, tr- scriptional responses, and temporary cell cycle arrest to allow the repair to take place. Defects in this system result in severe genetic disorders involving tissue degeneration, sensitivity to speci?c damaging agents, immunode?ciency, genomic instability, cancer predisposition and premature aging. The ?nding that many of the crucial players involved in DNA damage response are structurally and functionally conserved in different species spurred discoveries of new players through similar analyses in yeast and mammals. We now understand the chain of events that leads to instantaneous activation of the massive cellular responses to DNA lesions. This book summarizes several new concepts in this rapidly evolving ?eld, and the advances in our understanding of the complex network of processes that respond to DNA damage.

Two decades have passed since the mechanisms of protein synthesis became well enough understood to permit the genetic modification oforganisms. An impressive amount of new knowledge has emerged from the new technology, but much ofthe promise of20years ago has notyet been fulfilled. In biotechnology, efforts to increase the yields of commercially valuable metabolites have been less successful than ex pected, and when they have succeeded it has often been as much from selective breeding as from new methods. The cell is more complicated than what is presented in the classical teaching of biochemistry, it contains more structure than was dreamed of 20 years ago, and the behaviour ofany systemofenzymes is more elaborate than can be explained in terms ofa single supposedly rate-limiting enzyme. Even if classical enzymology and meta bolism may have seemed rather unfashionable during the rise ofmolecular biology, they remain central to any modification ofthe metabolic behaviour oforganisms. As such modification is essential in much ofbiotechnology and drug development, bio technologists can only ignore these topics at their peril."

A team of expert investigators and clinical researchers comprehensively review complement's basic biology, its role in disease, methods to measure its activity, and strategies for its inhibition in patients. Each chapter focuses on a specific area of basic and applied complement biology, spelling out the activation pathways and complement receptors. Informative animal models are discussed in detail, including the relative values of each model and the important interspecies differences that can distort the interpretation of preclinical studies. The emphasis throughout is on the pros and cons of the therapeutic use of recombinant complement inhibitors in specific diseases. Cutting-edge and innovative, Therapeutic Interventions in the Complement System highlights for today's researcher and biotechnologist effective strategies of drug discovery and development that are producing valuable new complement inhibitors for the treatment of a wide variety of clinically important diseases.

This comprehensive book explores the role of cytokines in immunotoxicology and human health using a variety of complex methods, from basic research to highly applied therapeutic applications. It includes a basic study of cytokines and details the effects of cytokines on the immune system and in treating cancer. The book serves as both a primer and a starting point for a more detailed investigation of the role these biological regulators play.

This book covers all aspects of oxygen delivery to tissue, including blood flow and its regulation as well as oxygen metabolism. Special attention will be paid to methods of oxygen measurement in living tissue and application of these technologies to understanding physiological and biochemical basis for pathology related to tissue oxygenation. This book is multidisciplinary and designed to bring together experts and students from a range of research fields including biochemical engineering, physiology, microcirculation, and hematology.

It has been known for a number of years that not only pathogenicity islands but also plasmids and bacteriophages are able to carry genes whose products are involved in pathogenic processes. Accordingly, such elements and their products play an important role in pathogenesis due to the intestinal E. coli as well due to Shigellae. Another interesting aspect which is reflected in different articles is that genomes evolve by acquisition of new pieces of DNA following gene transfer, but also by genome reduction. Different mechanisms include the deletion of sequences or the elimination of functions by the accumulation of point mutations or rearrangements.

Antibiotics are truly miracle drugs. As a class, they are one of the only ones that actually cure disease as opposed to most drugs that only help relieve symptoms or control disease. Since bacteria that cause serious disease in humans are becoming more and more resistant to the antibiotics we have today, and because they will ultimately become resistant to any antibiotic that we use for treatment or for anything else, we need a steady supply of new antibiotics active against any resistant bacteria that arise. However, the antibiotics marketplace is no longer attractive for large pharmaceutical companies, the costs of development are skyrocketing because of ever more stringent requirements by the regulatory agencies, and finding new antibiotics active against resistant strains is getting harder and harder. These forces are all combining to deny us these miracle drugs when we need them the most. I provide a number of possible paths to shelter from this perfect storm.

The Endoplasmic Reticulum (ER) is an organelle with extraordinary signaling and homeostatic functions. It is the organelle responsible for protein folding, maturation, quality control and trafficking of proteins destined for the plasma membrane or for secretion into the extracellular environment. Failure, overloading or malfunctioning of any of the signaling or quality control mechanisms occurring in the ER may provoke a stress condition known as ER stress . Accumulating evidence indicates that ER stress may dramatically perturb interactions between the cell and its environment, and contribute to the development of human diseases, ranging from metabolic diseases and cancer to neurodegenerative diseases, or impact therapeutic outcome. This book primarily focuses on the pathophysiology of ER stress. It introduces the molecular bases of ER stress, the emerging relevance of the ER-mitochondria cross-talk, the signaling pathways engaged and cellular responses to ER stress, including the adaptive Unfolded Protein Response (UPR), autophagy as well as cell death. Next the book addresses the role of ER stress in physiology and in the etiology of relevant pathological conditions, like carcinogenesis and inflammation, neurodegeneration and metabolic disease. The last chapter describes how ER stress pathways can be targeted for therapeutic benefit. Altogether, this book will provide the reader with an exhaustive view of ER stress biology and the latest insights in the role of ER stress in relevant human diseases."

Contents. List of Contributors. Brian Henderson and Gerry Higgs: Targets for modulating cytokine responses in inflammatory and infectious diseases. Mary Lee MacKichan and Anthony L. DeFranco: Cell signalling and cytokine induction by lipopolysaccharide. Rodger A. Allen and Stephen E. Rapecki: Regulation of cytokine production by inhibitors of cell signalling. Stanley T. Crooke: Oligonucleotide-based drugs in the control of cytokine synthesis. Peter I. Croucher, Ingunn Holen and Philip G. Hargreaves: Inhibiting cytokine-processing enzymes. Amanda Suitters and Roly Foulkes: Cytokine-neutralizing therapeutic antibodies. Ravinder N. Maini: The debut of anti-TNF therapy of rheumatoid arthritis in the clinic. Anthony Meager: Blockade of cytokine activity by soluble cytokine receptors. Michael F. Smith Jr.: Interleukin-1 receptor antagonist. Raymond J. Owens and Simon Lumb: Therapeutic regulation of cytokine signalling by inhibitors of p38 mitogen-activated protein kinase. Christian Bogdan, Yoram Vodovotz and John Letterio: TGF-ss and IL-10: inhibitory cytokines regulating immunity and the response to infection. Brian Henderson: Therapeutic control of cytokines: lessons from microorganisms. Index

This book covers lymphoproliferative disorders in patients with congenital or acquired immunodeficiencies. Acquired immunodeficiencies are caused by infections with the human immunodeficiency virus or arise following immunosuppressive therapy administered after organ transplantation or to treat connective tissue diseases such as rheumatoid arthritis. It was recently discovered that various diseases or therapeutic modalities that induce a state of immunosuppression may cause virally driven lymphoproliferations. This book summarizes for the first time this group of immunodeficiency-associated lymphoproliferations.

These proceedings contain selected contributions from the participants to the Fourth International Symposium on Dendritic cells that was held in Venice (Lido) Italy, from Oc tober 5 to 10, 1996. The symposium was attended by more than 500 scientists coming from 24 different countries. Studies on dendritic cells (DC) have been greatly hampered by the difficulties in preparing sufficient cell numbers and in a reasonable pure form. At this meeting it has been shown that large quantities of DC can be generated from precursors in both mice and humans, and this possibility has enormously encouraged studies aimed to characterize DC physiology and DC-specific genes, and to employ DC therapeutically as adjuvants for im munization. The possibility of generating large numbers of autologous DC that can be used in the manipulation of the immune response against cancer and infectious diseases has tremendously boosted dendritic cell research and the role of DC in a number of medi cal areas has been heatedly discussed."

Cancer Vaccines: Methods and Protocols explores the manipulation and modification of immune cells, the manipulation and modification of tumor cells as well as the manipulation of immune/tumor interactions and various delivery mechanisms, with the overall end goal of evoking a tumor-specific response and overcoming the immuno-evasive mechanisms employed by the tumor cells. This detailed volume also covers the subject of cancer vaccines in a more global sense with its section on the advances, challenges, and future of cancer vaccines. 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. Comprehensive and authoritative, Cancer Vaccines: Methods and Protocols aims to help guide researchers toward developing further generations of cancer vaccines that are both safe and efficacious, with the hope that cancer vaccines will be the standard of care in the very near future.

In the post genomic era, understanding of the innate immune system is enriched by findings on the specificity of innate immune reactions as well as to novel functions that do not strictly correlate with immunological defense and surveillance, immune modulation or inflammation. This volume covers natural killer cells, mast cells, phagocytes, toll-like receptors, complement, host defense in plants and invertebrates, evasion strategies of microorganisms, pathophysiology, protein structures, design of therapeutics, and experimental approaches.

The three years since our last conference in San Francisco have again seen a dramatic expansion of the number of antivirals either licensed or in the late stages of clinical trials. d4T is now licensed for HIV infection, famciclovir and the oral pro-drug of acyclovir, valacyclovir, are now licensed for VZV infections in some countries. Moreover. oral ganciclovir, cidofovir, and sorivudine are not far behind. Clinical trials with the second-site reverse transcriptase inhibitors and the protease inhibitors for HlV infection are proceeding rapidly and on a broad scale, and the preliminary results would suggest that several of these classes of drugs will be licensed as well. Despite this optimism, however, there is increasing evidence that antiviral-resistant strains of pathogenic viruses will be a significant problem, perhaps especially with therapy of HIV infection, and there remains a desperate need for improved drugs (with either improved efficacy or decreased toxicity, or both) for CMV and HIV infections. This book is the edited proceedings of the Fourth Triennial Conference on Antiviral Chemotherapy, held in San Francisco, in November 1994. The conference was sponsored by the University of California, San Francisco, and co-sponsored by the International Society for Antiviral Research (ISAR), the Macfarlane Burnet Centre for Medical Research in Melbourne, Australia, and the Australian National Centre for HIV Virology Research. The conference had been organized to present an overview of the field of antiviral chemotherapy.

TLR4 is one of the most important innate immunity receptors, its function mainly consisting in the activation of inflammatory pathways in response to stimulation by Pathogen-Associated Molecular Patterns (PAMPs) and Damage Associated Molecular Pattern molecules (DAMPs). This volume critically reviews the different types of TLR4 activators and inhibitors, discusses the role of molecular aggregates in agonism/antagonism as well as the pivotal role of the CD14 receptor in the modulation of TLR4 signal and the molecular details and actors of the intracellular cascade. The book presents the role of TLR4 in several pathologies, such as sepsis and septic shock caused by receptor activation by gram-negative bacterial lipopolysaccharide (LPS), in neurodegenerative and neurological diseases such as Parkinson and Alzheimer's diseases, and Amyotrophic Lateral Sclerosis (ALS). It reviews the role of TLR4 in neural stem cell-mediated neurogenesis and neuroinflammation and in Human Induced Pluripotent Stem Cells and Cerebral Organoids and discusses the emerging role of micro-RNA (miRNA) regulation by TLR4.

Neutrophils, the most abundant white cells in humans, serve as the primary cellular defense against infection. Neutrophil Methods and Protocols, Second Edition provides a concise set of protocols written by leading researchers in the field for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step diagnostic assays of common neutrophil disorders. Topics covered include an overview of neutrophils and their role in host defense and inflammation; methods most commonly used for isolating neutrophils from humans and other animal species; procedures for subcellular fractionation of human neutrophils, analysis of in vivo transmigrated neutrophils, generation of mature neutrophils from induced pluripotent stem cells and analysis of neutrophil gene expression; methods for investigating priming, oxidant production, phagocytosis, bactericidal activity and extracellular trap formation and protocols for investigating neutrophil adhesion, chemotaxis and outside-in signaling via integrins. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Neutrophil Methods and Protocols, Second Edition is a comprehensive source for detailed explanations and applications of the most modern methodological advances in neutrophil biology. Both basic scientists and clinicians will find a collection of this caliber to be an invaluable aid in their work with neutrophils.