This book provides basic, simple, and logical explanations for choices to be made to run the best Elispot possible. It allows the newcomer to truly understand the best options for specific protocol steps, reagents and materials, and provides even the experienced Elispot user with insight into best practices. The techniques presented here are supported by the author's twenty-plus years of first-hand experience working with this assay, as well as the shared experiences of numerous colleagues and collaborators. The enzyme-linked immunospot (Elispot) assay is a widely used technique to monitor cells on the single cell level for the release of analytes like cytokines, chemokines or immunoglobulins (antibodies), in response to particular stimuli. The most important feature of Elispot is its outstanding sensitivity, allowing the detection of specific cells in very low frequencies. The advantages of Elispot have resulted in it being widely adapted for use in research and translational applications in numerous fields including cancer, infectious diseases, autoimmunity, and transplantation.

This book provides a comprehensive overview of the cascade of events activated in the body following the implant of biomaterials and devices. It is one of the first books to shed light on the role of the host immune response on therapeutic efficacy, and reviews the state-of-the-art for both basic science and medical applications. The text examines advantages and disadvantages of the use of synthetic versus natural biomaterials. Particular emphasis is placed on the role of biomimicry in the development of smart strategies able to modulate infiltrating immune cells, thus reducing side effects (such as acute and chronic inflammation, fibrosis and/or implant rejection) and improving the therapeutic outcome (healing, tissue restoration). Current cutting-edge approaches in tissue engineering, regenerative medicine, and nanomedicine offer the latest insights into the role immunomodulation in improving tolerance during tissue transplant in the treatment of orthopaedic, pancreatic, and hepatic diseases. "Immune Response to Implanted Materials and Devices" is intended for an audience of graduate students and professional researchers in both academia and industry interested in the development of smart strategies, which are able to exploit the self-healing properties of the body and achieve functional tissue restoration.

The latest in a series of books from the International Hypoxia Symposia, this volume spans reviews on key topics in hypoxia, and abstracts from poster and oral presentations. The biannual International Hypoxia Symposia are dedicated to hosting the best basic scientific and clinical minds to focus on the integrative and translational biology of hypoxia. Long before 'translational medicine' was a catchphrase, the founders of the International Hypoxia Symposia brought together basic scientists, clinicians and physiologists to live, eat, ski, innovate and collaborate in the Canadian Rockies. This collection of reviews and abstracts is divided into six sections, each covering new and important work relevant to a broad range of researchers interested in how humans adjust to hypoxia, whether on the top of Mt. Everest or in the pulmonary or cardiology clinic at low altitude. The sections include: Epigenetic Variations in Hypoxia High Altitude Adaptation Hypoxia and Sleep Hypoxia and the Brain Molecular Oxygen Sensing Physiological Responses to Hypoxia

This volume provides current methods and protocols for gene and protein delivery based on both lentivirus-generated and spontaneously released nanovesicles. Lentiviral Vectors and Exosomes as Gene and Protein Delivery Tools guides readers through methods on macromolecule delivery and chapters describe the LV-based protocols of gene engineering. 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 cutting-edge, Lentiviral Vectors and Exosomes as Gene and Protein Delivery Tools aims to ensure successful results in the further study of this vital field.

This book provides readers an extensive overview of recent progress in basic and clinical research on cancer immunotherapy. Thanks to rapid advances in molecular biology and immunology, it has become increasingly evident that cancer growth is influenced by host immune responses. With the success of a number of clinical trials, immunotherapy has become a promising treatment modality of cancer. This book covers five major topics, including monoclonal antibodies, biological response modifiers, cancer vaccines, adoptive cellular therapy and oncolytic viruses. It also examines the combination of different immune strategies as well as the combination of immunotherapy with other treatments to increase anti-tumor effects. Through the comprehensive discussion of the topic, the book sheds valuable new light on the treatment of tumors.

This volume reviews the current state of research on the IgM antibody and its multiple receptors and binding proteins. Interactions of the IgM ligands with these molecules are important for protection against infections as a first line of defense, as well as for regulation of immune responses to pathogens and self-antigens. The book includes up-to-date information on: (i) the phylogeny of three IgM-binding receptors [polymeric Ig receptor (pIgR), Fc receptor for both IgA and IgM (Fc /R), and Fc receptor for IgM only (FcR)]; (ii) the lymphocyte-restricted distribution and unique ligand-binding activity of FcR; (iii) the definition and potential function of Fc receptor-like molecule A (FCRLA) as a resident endoplasmic reticulum protein that binds IgM, but also IgG and IgA; (iv) IgM antibody-mediated enhancement of humoral immune responses, highlighting the importance of complement and its receptors, (v) the numerous important roles of IgM natural antibodies in regulation of inflammation. It is an invaluable resource for researchers and clinicians alike.

This book explores the major cytokines, such as IL-1 and IFN- , with respect to the regulation of their gene expression and protein production in specific immune cell types. It discusses both healthy physiological settings and in pathological situations in which the expression of some cytokines could be dysregulated, resulting in either immunodeficiency or exacerbated inflammatory sequelae in animal models as well as in human patients. Cytokines are important regulators of immune responses that require the highly coordinated participation and communication of multiple cell types. The expression of cytokines by various producer cell types is therefore carefully regulated in response to environmental cues at multiple levels: transcription, translation and posttranslational modification. Presenting cutting-edge advances in our understanding of the regulation of cytokine expression, this book is a valuable resource for anyone involved or interested in immune regulation.

This second edition volume provides detailed protocols that address the challenges of signal-transduction IHC. This book delves into chapters that discuss the nature of signal transduction phenomena and approaches to making phosphor-specific antibodies, as well as numerous bona fide methods methods on digital imaging techniques, preservation of tissue targets, multicolor detection, flow cytometry, lipophagy analysis, apoptosis, and the combination of IHC with in situ hybridization. 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 comprehensive, Signal Transduction Immunohistochemistry: Methods and Protocols, Second Edition is a valuable resource to both novices and experts in other fields of biomedical research who need advice on IHC protocols to study signal transduction. This book will also be useful for researchers in academia, government labs, and the biotech industry.

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.

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.

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.

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.

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.

The concept of immunotherapy was in infancy when the first edition was written; since then, major advances have been made, not only with several prominent clinical trials, but also with the approval of cell-based therapy by the FDA for the treatment of cancer in 2010. These events resulted in a gradually narrowing gap between early scientific knowledge and the late development of immune-based therapies. Consequently, the significance and magnitude of these advances warranted a revision of this contribution; this revised edition will provide a deeper understanding of the recent advances and discoveries related to the function of the immune response and their applications in the development of novel therapies to treat human diseases. Some of the key discoveries during the past five years include: the identification of the new subsets of helper T cells; new cytokines and their networks; and novel signal transduction mechanisms. For example, the identification of TH17 subset of helper T cells, in addition to TH1 and TH2 cells, not only advanced our understanding of the function of the basic immune response, but also raised our awareness of the possible etiology and pathogenesis of diseases such as allergy, asthma, rheumatoid arthritis, and other auto-immune/immune system based diseases. The newly identified powerful cytokine networks, that regulate both innate and acquired immune responses, emerged as a result of the finding of new cell types such as innate lymphoid cells and iNKT. Identification of the novel cytokines and their networks has advanced our knowledge of the mechanisms involved in the maintenance of tissue homeostasis, including inflammation and tissue repair during stress and injury. The development of HIV vaccines has also seen dramatic changes over the last few years. There has been a shift from a sole focus on T cell vaccines to a holistic approach that pertains to the induction of both humoral and cellular elements. This entails the induction of antibodies - both binding and neutralizing - to prevent infection. The cellular vaccination produces a safety net of CD8+ T-cell responses to suppress the replication of the virus in the infected patients, and both of the effector arms are aided by helper T cells. From the perspective of clinical applications, significant advances have also been made in: oral immunotherapy for allergic disease, the possible treatment of HIV infection, the development of new monoclonal antibodies and their fragments to treat human diseases, and immune cell based therapies for cancer.

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.

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 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.

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 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.

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 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.

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.

NEW YORK TIMES BESTSELLER Tabitha Brown, the #1 New York Times bestselling author of Feeding the Soul, presents her first cookbook-full of easy, family-friendly vegan recipes and stories from the spirit, inspired by her health journey and love of delicious food. Sometimes people say to Tabitha Brown, "I've never eaten vegan before." As Tab says, "Have you ever eaten an apple?" After living with a terrible undiagnosed illness for more than a year and a half, Tab was willing to try anything to stop the pain. Inspired by the documentary What the Health, she tried a thirty-day vegan challenge-and never looked back. Wanting to inspire others to make changes that might improve their own lives, she started sharing her favorite plant-based recipes in her signature warm voice with thousands, and now millions, of online fans. Tab's recipes are flexible, creative, and filled with encouragement, so you trust yourself to cook food the way it makes you happy. If you're already a "cooking from the spirit" sort of person, you'll love how much freedom Tab gives to make these delicious vegan dishes your own. If you're newer to cooking-or to vegan cooking-Tab will help you get comfortable in the kitchen and, most important, have fun doing it! In this joyful book, Tab shares personal stories, inspirational "Tabisms," and more than eighty easy, family-friendly recipes, including: Yam Halves Topped with Maple-Cinnamon Pecan Glaze Stuffed Avocado Jackfruit Pot Roast Crab-less Cakes with Spicy Tartar Sauce Who Made the Potato Salad? Kale and Raspberry Salad Strawberry Cheesecake Cups Cooking from the Spirit is for anyone interested in plant-based eating and all lovers of food, plus anyone who wants a little warm inspiration in their lives. As Tab says, "Honey, now let's go on and get to cooking from the spirit. Yes? Very good!"

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.