This volume provides readers with methods and protocols for understanding the development of recombinant viruses and their use as vaccines platforms. Recombinant Virus Vaccines: Methods and Protocols details the use of recombinant vaccines that are employed to either produce immunogens in vitro or elicit antibody production in vivo. The chapters in this book are divided into four parts: Part I explores double-stranded DNA viruses; Part II discusses negative sense single-stranded RNA viruses; Part III talks about positive sense single-stranded RNA viruses; and Part IV describes bacteriophages. 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. Thorough and cutting-edge, Recombinant Virus Vaccines: Methods and Protocols is a valuable resource for scientists and clinicians who are interested in learning about and adopting methods for use in basic and biomedical research directed toward generating and developing recombinant viral vaccines.

This book contains the refereed contributions from the 43rd annual meeting of ISOTT. The annual meetings of ISOTT bring together scientists from various fields (medicine, physiology, mathematics, biology, chemistry, physics, engineering, etc.) in a unique international forum. ISOTT conferences are a place where an atmosphere of interaction is created, where many questions are asked after each presentation and lively discussions occur at a high scientific level. This vivid interaction is the main motivation for members to participate and gain new ideas and knowledge in the broad field of oxygen transport to tissue. The proceedings include sessions covered various research topics including Multi-Modal Imaging/Spectroscopy & Instrumentation; Cancer Metabolism; Cellular Hypoxia and Mitochondrial Function; Brain Oxygenation and Function; Other Organ Function and Metabolism; Oxygen Transport in Sports, Diseases and Clinical Care; Acupuncture, Meridians, and Primo Vascular System; EPR, MRS and MRI.

This book provides a comprehensive overview of the cutting-edge discoveries and the systems approaches related to the multi-disciplinary field of psychoneuroimmunology (PNI), which may provide the scientific basis for mind-body relationships, and aid in the development of personalized medicine. Specifically, the book focuses on the systems biology studies of complex mechanisms, such as stress and inflammation. There is strong evidence supporting the close relationship between stress, inflammation, and disorders including obesity, cardiovascular disease, diabetes, skin diseases, and sleep disorders. As inflammation is a critical connection among different diseases, further elucidating this connection may contribute to the findings of systemic therapeutic targets. Translational medicine is also addressed, by applying PNI approaches in clinical practice. A major challenge in current bioscience is the translation of basic scientific discoveries into better clinical outcomes. With the understanding of the translational implications of PNI, integrative interventions can be applied to modulate stress responses and to promote healthier behaviors. As an important feature of this book, a broad overview of the PNI-associated integrative interventions is provided. These interventions include nutritional supplements, meditation, and other mind-body strategies. From basic concepts to systems methodologies, from theoretical sources to clinical applications, Psychoneuroimmunology contributes to the understanding and development of PNI toward the establishment of personalized and mind-body medicine.

In spite of their relatively low prevalence, systemic vasculitides have been the object of intensive basic and clinical investigations over the last few years. As a consequence, important advancements have been achieved: from updated diagnostic and classification criteria and a more rational nomenclature to the recognition of an expanding spectrum of clinical manifestations and potentially devastating complications; from the recognition of the viral etiology of conditions such as HCV-related cryoglobulinemic vasculitis and HBV-associated polyarteritis nodosa to newly named variable vessel vasculitis exemplified by Behcet's disease; from single-organ vasculitis such as central nervous system vasculitis to the more recently emerging features of the IgG4-related, immune-mediated diseases that are showing remarkable clinical heterogeneity. In addition, intriguing pathogenetic hypotheses are being reported for certain chronic, relapsing vasculitides that are improving our understanding of their biology and basic pathophysiology. New avenues are being explored that will hopefully allow a deeper comprehension of the relationships between certain virus-driven vasculitides and lymphoproliferation, and possibly lead to the identification of novel biomarkers that may be used to single out patients at an increased risk of relapse. This explosion of knowledge is obviously resulting in state-of-the-art, personalized treatments of systemic vasculitides. This book is a collection of reviews on the major vasculitides, written by scientists and clinicians with a multi-year experience in this field. We hope it will provide the reader with a stimulating container of new advances in scientific knowledge and more rational therapeutic approaches to this fascinating chapter of pathology.

Currently, malaria kills more than 300,000 people per year, making it a top priority of world leaders and international organizations, who are working towards implementing and coordinating efforts to eradicate this disease. An effective malaria vaccine is recognized as the key element that will decide between success and failure in this fight. At present, despite intensive research efforts, such a vaccine is not yet available for use. However, there are a number of advanced candidate vaccines with high chances of success in the short term. Malaria: Immune Response to Infection and Vaccination provides a comprehensive view on the immune response to malaria and to the different vaccines under development. The book offers the following: - Contributions by top research leaders in the field, - Comparisons of the immune responses to both malaria infection and malaria vaccines, which are traditionally treated separately, - Coverage of the immune responses to the different stages of malaria, which are frequently treated as separate fields of research.

Second comprehensive volume focuses on anti-inflammatory nutraceuticals and their role in prevention and therapy of various chronic diseases. Food and drug administration (FDA) approved drugs such as steroids, non-steroidal anti-inflammatory drugs (NSAIDS), statins and metformin have been shown to modulate inflammatory pathways, but their long-term intake has been associated with numerous side effects. Thus dietary agents which can modulate inflammatory pathways in humans, are likely to exhibit enormous potential. Leading experts describe the latest results of anti-inflammatory nutraceuticals and their role in prevention and therapy of various chronic diseases.

This volume focuses on recent advances in understanding T cells as key players in antitumor immune responses, and as a result T cell-based immunotherapy is starting to transform the treatment of advanced cancers. However, despite recent successes, many patients with cancer fail to respond to these treatments. Defective migration of T cells into and within tumors is considered as an important resistance mechanism to cancer immunotherapy.The volume includes three sections. The first section covers general knowledge about T cell trafficking during a normal immune response but also during tumor development. The second section provides an in-depth description of the different obstacles that prevent T cells from migrating and contacting tumor cells. The third section explores therapeutic strategies to improve trafficking of T cells into tumors and, thus, to enhance the effectiveness of cancer immunotherapy.

Aimed at both beginners and experienced researchers unfamiliar with the technique, this book provides a concise and easy-to-understand guide to using immunohistochemistry. It identifies the proper place and purpose of each component of immunohistochemistry, emphasizes the components that are of critical importance, and explains the logistics of experimental approaches. Understanding the principles of immunohistochemistry allows for the identification and localization of protein targets and is of crucial use in the following experiments: * producing transgenic animals * studying the effects of drug treatments * electrophysiological studies * animal surgery * isolating and differentiating stem cells * super-resolution microscopy Immunohistochemistry for the Non-Expert is aimed at a wide audience, including molecular biologists, pharmacologists, immunologists, chemists, cell biologists, physiologists, and researchers involved with animal anatomy studies.

This book describes modern biophysical techniques that enable us to understand and examine dynamic processes of infection at the molecular level. Cutting-edge research articles, laboratory protocols, case studies and up-to-date reviews cover topics such as single-molecule observation of DNA replication repair pathways in E. coli; evolution of drug resistance in bacteria; restriction enzymes as barriers to horizontal gene transfer in Staphylococcus aureus; infectious and bacterial pathogen biofilms; killing infectious pathogens through DNA damage; bacterial surfaces in host-pathogen interactions; bacterial gene regulation by riboswitches; transcription regulation in enterobacterial pathogens; the bacterial flagellar motor; initial surface colonization by bacteria; Salmonella Typhi host restrictions; as well as monitoring proton motive force in bacteria; microbial pathogens using digital holography; mathematical modelling of microbial pathogen motility; neutron reflectivity in studying bacterial membranes; force spectroscopy in studying infection and 4D multi-photon imaging to investigate immune responses. The focus is on the development and application of complex techniques and protocols at the interface of life sciences and physics, which increase the physiological relevance of biophysical investigations.

This volume contains collection of Natural Killer Cell methodologies relevant for both basic and translational research. These methodologies present new developments in the natural killer (NK) cell field, such as understanding the influence of NK cells metabolism on its function, identifying complexity of NK cell subsets through mass cytometry, and determining the emergence of memory NK cells in murine model of MCMV infection. Methods that study NK cell migration and cytotoxicity through endpoint analysis or live single cell imaging are also discussed. Chapters also describe methods pertaining to translational application of NK cells, such as ex vivo expansion of NK cells on K562 cell lines genetically modified to express either membrane bound IL-15 or membrane bound IL-21, large scale NK cell culture, current techniques for engineering NK cells to express chimeric antigen receptors or chemokine receptors using retroviral vectors, electroporation of mRNA, and the natural phenomenon of trogocytosis. Written in the highly successful Methods in Molecular Biology series format, these 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. Cutting edge and thorough, Natural Killer Cells: Methods and Protocols is a valuable resource for researchers who not only want to understand mechanisms that govern NK cell behavior and diversity, but also for those who want to understand how to systematically evaluate NK cells for adoptive immunotherapy applications.

This volume covers the topics presented at the 3rd International Conference on Tumor Microenvironment and Cellular Stress by an international community of researchers. The conference brings together scientists to discuss different cellular and animal models of tumor microenvironment study and identify common pathways that are candidates for therapeutic intervention; stimulate collaboration between groups that are more focused on elucidation of biochemical aspects of stress biology (e.g., HIF regulation) and groups that study the pathophysiological aspects of stress pathways or engaged in drug discovery; and critically evaluate novel targets for imaging or therapeutic intervention that would be of use to the tumor microenvironment community and pharmaceutical industry.

This volume brings together recent developments in quasispecies theory extended to variable environments and practical applications in elucidating viral dynamics and treatment designs. In particular, the existence of an error threshold in rugged fitness landscapes has opened the way to a new antiviral strategy termed lethal mutagenesis, which is now under intensive theoretical, experimental and clinical investigation. As such the book explains how an understanding of quasispecies dynamics within infected organisms has increased our knowledge of viral disease events. From a clinical perspective, population dynamics highlights important problems for viral disease control, such as the selection of drug-resistant mutants that often accompanies treatment failures, and suggests means of increasing the effectiveness of antiviral treatments. The book is intended for students and scientists interested in basic and applied aspects of biophysics, chemistry, biology, evolution and medical virology.

This book offers an overview of our current understanding of host defense peptides and their potential for clinical applications as well as some of the obstacles to this. The chapters, written by leading experts in the field, detail the number and diversity of host defense peptides, and discuss the therapeutic potential not only of antibacterial, but also of antifungal, antiviral, plant antimicrobial and anticancer host defense peptides. The authors provide new insights into their mechanisms of action and their immunomodulatory properties, and review recent advances in the design of novel therapeutic molecules. Lastly, their potential to prevent preterm births and Staphylococcus aureus infections is highlighted. The book is of interest to researchers, industry and clinicians alike.

Cancer care is undergoing a radical transformation as novel technologies are directed toward new treatments and personalized medicine. The most dramatic advances in the treatment of cancer have come from therapeutics that augment the immune response to tumors. The immune checkpoint inhibitors are the best-known and most highly advanced examples of Immune Therapeutics targeting tumor cells and include approved antibody drugs directed at the cell surface proteins CTLA4 and PD-1. These are now considered foundational treatments for several solid tumor indications, and that list of indications is growing quickly. More broadly, antibodies have become workhorse molecules across the entire immunotherapy landscape. Antibodies to novel targets modulate the activity of diverse immune cell regulatory proteins. Engineered antibodies can induce tumor cell death or expose tumor cells to poisonous toxins (ADCC and ADC, respectively). Bi-specific antibodies can engage multiple tumor targets simultaneously, or can redirect lymphocytes to attack tumor cells. The antigen-binding domains within antibodies can be spliced onto cell stimulatory domains and transduced into T cells or NK cells, creating remarkable tumor-specific cellular therapeutics (CAR-T, CAR-NK). Beyond antibody-based therapies there are highly diverse and differentiated technology tool kits being applied to immunotherapy. Small molecule drugs are being developed to attack the tumor microenvironment, novel tumor vaccine approaches are showing great promise, patient lymphocytes are being isolated, expanded and reintroduced to patients, gene-editing techniques are becoming widely deployed, and a vast number of new tumor targets, and mutated tumor proteins (neoantigens), are being discovered. The past decade has seen unprecedented success in the treatment of diverse cancers. The authors of this volume have been asked to not only review progress to date, but importantly, to look ahead, and anticipate the evolution of cancer treatment across diverse Immune Therapeutic approaches. Our hypothesis is that the advances we are seeing across the immunotherapy landscape will further evolve and synergize, leading us finally to outright cures for many cancers.

This second edition volume expands on the first edition with new developments on Toll-Like Receptors (TLRs) controlling events such as cross-priming of associated pattern recognition receptors, post-transcriptional regulation, interaction with other cellular and biologic systems, and cancer progression. This book is divided into five sections: Part I outlines methods for TLR detection, interaction, and intracellular trafficking; Part II describes methods and assays to investigate how TLRs cross-prime other pattern recognition receptors, including intracellular DNA receptors and inflammasome formation; Part III highlights RNA regulation, detailing how TLRs can induce RNA transcripts and molecules such as lncRNAs and microRNAs; Part IV explores TLR detection and activation in systems such as epithelial barrier function, metabolism and the circadian clock, as well as cellular systems including T and B lymphocytes; and Part V describes models to delineate the role of TLRs in diseases such as dermatitis, arthritis, and gastric cancer. Written in the highly successful Methods in Molecular Biology series format, each chapter contains a summary, a list of required materials, step-by-step, readily reproducible laboratory protocols, useful notes to investigate TLRs in cell culture, systems and disease, and tips on troubleshooting and avoiding known pitfalls. Practical and cutting-edge, Toll-Like Receptors: Methods and Protocols, Second Edition is a valuable resource to any immunologist, molecular or medical biologist working in a laboratory setting. It will add skill to both students and the more advanced molecular biologist who wishes to learn a new technique or move to a different area within their current repertoire of practical knowledge.

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.

This thesis describes the use of biophysical and biochemical methods to prove that calcium has a positive feedback effect on amplifying and sustaining CD3 phosphorylation and should enhance T-cell sensitivity to foreign antigens. The study presented shows that calcium can regulate the signal pathway in cells not only as a secondary messenger but also through direct interactions with the phospholipid bilayer. The approach used in the thesis also represents an important advance, as it couples the use of nuclear magnetic resonance (NMR) to the analysis of signaling phenomena in living cells. Moreover, the thesis optimizes the Nanodisc assembly protocol, which can broaden its range of applications in membrane protein studies. A preliminary study on the structure of dengue virus NS2B-NS3p in complex with aprotinin, which may help to develop new drugs against the dengue virus, is also included.

This book describes the molecular biology, pathogenesis, epidemiology, and potential strategies for control of chikungunya virus (CHIKV) infection. It offers insight into the structure and functions of CHIKV proteins as they relate to host response, interaction with the arthropod vector, and vaccination. A detailed account of both the epidemiological outlook and the clinical syndrome of CHIKV infection is provided. The complex host-virus interaction and the signaling pathways that mediate such interactions are also covered. Throughout the book, graphics and charts are used to provide stimulating discussion on important findings in the field of chikungunyalogy. The chapters are written with a global perspective by experts of CHIKV from around the world. This project is especially significant given that CHIKV is a pathogen of worldwide public health concern. Although the presence of CHIKV infection is not global yet, worldwide dissemination is predicted in the future due largely to the lack of effective treatment/therapy, efficient control of transmission, and knowledge about mechanisms of pathogenesis. Additionally, globalization of CHIKV is predicated on its mode of dissemination (mosquito vector) and cross border travel and migration.

This volume focuses on the roles of long non-coding RNAs (lncRNAs) in contexts ranging from human cancers to cardiovascular disease and ageing. The role of lncRNAs in X-inactivation and those lncRNAs derived from pseudogenes, past retroelements integrated within the human genome, as well as the role these pseudogene-derived lncRNAs play in cancer development are discussed in detail. Further, the book examines the function of lncRNAs in diseases such as diabetes, in smooth muscle formation, and in the modulation of nuclear receptors, as well as in connection with perspectives on the development of personalized therapeutics. It offers an appealing and insightful resource for scientists and clinicians alike.

This text provides a practical guide providing step-by-step protocol to design and develop vaccines. Chapters detail protocols for developing novel vaccines against infectious bacteria, viruses, fungi, and parasites for humans and animals. Volume 1: Vaccines for Human Diseases has an introductory section on how vaccines impacted diseases, the immunological mechanism of vaccines, future challenges for vaccinologists, and current trends in vaccinology. The design of human vaccines for viral, bacterial, fungal, parasitic and prion diseases as well as vaccines for drug abuse, allergy, and tumor vaccines are also described in this volume. As a volume in the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Vaccine Design: Methods and Protocols, Volume 1: Vaccines for Human Diseases aims to ensure successful results in the further study of this vital field.

The present volume focuses on microbial invasion strategies of pathogen uptake. An accompanying volume (Vol. 5) in the series presents the phagocytic process from the viewpiont of the host cell.
This field of study is growing rapidly after a somewhat slow start over recent decades. This collection of invited chapters attempts to reflect current research and brings together cell biologists, microbiologists, and immunologists wiht disthemes, hopefully like a symphony rather than a boring catalogue. It will be evident that editorial bias favors intracellular parasitism and medically important organisms. The neutrophil is far more than a supporting player to the macrophage and some attempt is made to remind the reader of some of its unique skills. To retain a manageable size, the emphasis is on relatively early events such as mutual recognition, cell entry, and response, rather than on longterm changes in gene expression by either host cell or pahtogen. Viruses are excluded not because of lack of importance but because of somewhat different research approaches, although it is becoming increasingly clear that large viruses (e.g. "Vaccinia") and "Listeria monocytogenes," share common strategies in invasion and intercellular spread.

Sabine Stubler compares different proteasome isoforms and subtypes in terms of their transport and active site-related parameters applying an existing computational model. In a second step, the author extends this model to be able to describe the influence of proteasome inhibitors in in vitro experiments. The computational model, which describes the hydrolysis of short fluorogenic peptides by the 20S proteasome, is calibrated to experimental data from different proteasome isoforms using an approximate Bayesian computation approach. The dynamics of proteasome inhibitors are included into the model in order to demonstrate how to modulate the inhibitor's transport parameters for strong or isoform-specific inhibition.

This book provides a comprehensive review of the structure, function and pathophysiology of the pulmonary vasculature. Emerging evidence reveals the multifaceted roles played by the pulmonary vasculature. To reflect those roles, the individual chapters address topics ranging from pulmonary blood vessel development to vascular endothelial apoptosis, and delve deeply into our current understanding of various aspects of the pulmonary vasculature.

This book focuses on the function of antibodies in vivo. Recent years have seen an exponential growth in knowledge about the molecular and cellular mechanisms of antibody activity. These new results dramatically changed our view of how antibodies function in vivo. The importance of this class of molecules is demonstrated by the heightened susceptibility to infections of humans and mice with an altered capacity to generate pathogen specific antibody responses. Thus, the majority of our currently available vaccines, such as vaccines against influenza, measles and hepatitis focus on the generation of long lasting antibody responses. Recent evidence from a variety of in vivo model systems and from human patient cohorts has highlighted the exclusive role of cellular Fc-receptors for certain immunoglobulin isotypes and subclasses. With the recent discovery of a human Fc-receptor for IgM all different human immunoglobulin isotypes now have a cellular receptor, providing a feedback mechanism and link between antibodies and the cellular components of the immune system. Moreover it has become clear the complement and Fc-receptor system are tightly connected and regulate each other to ensure a well balanced immune response. Among the immunoglobulin isotypes IgG plays a very important protective role against microbial infections and also as a therapeutic agent to kill tumor cells or autoantibody producing B cells in autoimmune disease. Transfer of our knowledge about the crucial function of Fc-receptors has led to the production of a second generation of therapeutic antibodies with enhanced binding to this class of receptors. Binding of antibodies to Fc-receptors leads to the recruitment of the potent pro-inflammatory effector functions of cells from the innate immune system. Hence, Fc-receptors link the innate and adaptive immune system, emphasizing the importance of both arms of the immune system and their crosstalk during anti-microbial immune responses. Besides this pro-inflammatory activity immunoglobulin G (IgG) molecules are long known to also have an anti-inflammatory function. This is demonstrated by the use of high dose intravenous immunoglobulins as a therapeutic agent in many human autoimmune diseases. During the past five years several new insights into the molecular and cellular pathways of this anti-inflammatory activity were gained radically changing our view of IgG function in vivo. Several lines of evidence suggest that the sugar moiety attached to the IgG molecule is responsible for these opposing activities and may be seen as a molecular switch enabling the immune system to change IgG function from a pro- to an anti-inflammatory activity. There is convincing evidence in mice and humans that aberrant IgG glycosylation could be an important new pathway for understanding the impaired antibody activity during autoimmune disease. Besides this tremendous increase in basic knowledge about factors influencing immunoglobulin activity the book will also provide insights into how these new insights might help to generate novel therapeutic approaches to enhance IgG activity for tumor therapy on the one hand, and how to block the self-destructive activity of IgG autoantibodies during autoimmune disease on the other hand.