Principles of innate and adaptive immunity xix 16-22 Isolation of lymphocytes. 739 700 A20 e seanmnegegtealpodhmpootesty 16-23 cting A-21 ympyesrmissusther than bld. 702 Flow cytometry and FACS analysis. 16-24 702 isationsingayoted maeti 16-25 strates the importance A-24 Isolation of homoceneous T-cell lines. 704 16-26 an 743 705 16-27 1 705 42 16-28 707 4-28 tetramers. 746 16-29 高 A29 &5esenethe.dersnyoftheToelepertoreby 748 16-30 708 A-30 749 1631 A-31 on by treatme ns 708 711 AesrgoPtyheTEL 5 Questions. A-34 Assays for CD4Tcells. 751 General references Section references. 712 Detection of immunity in vivo 753 A36 ntoiprote Appendix I The Immunologist's Toolbox 717 A37 The tuberculin test. mmuntaion 717 esingrallergics 含 A-2 728 Aneeegmnsporssandmunoged 高 A40 The Arthus reaction. A4 Adjuvants. A43 hemao0Oe The detection,meas ofTcelsansies rement,and characterization A44 hirsh and A-45 Transoenic mice. A-46 Gene knockout by targeted disruption. 723 Hemagglutinaion and bood typing Appendix Il CD Antigens 763 Appendix l Cytokines and Their Receptors A-9 9 measurementof antbody affinity 727 Appendix IV Chemokines and Their Receptors 782 4-10 Anti-immunoglobuin antibodes. Biographies 784 12 Photograph Acknowledgments 785 A-13 731 Micscopy and imagng. 7 Glossary 18 A.14 A-15 Immunoelectron microscopy. Index 823 734 A-16 Immunohistochemistry. 9enesandiherprodlucs
16-22 Live-attenuated viral vaccines are usually more potent than 'killed' vaccines and can be made safer by the use of recombinant DNA technology. 700 16-23 Live-attenuated vaccines can be developed by selecting nonpathogenic or disabled bacteria or by creating genetically attenuated parasites (GAPs). 702 16-24 T he route of vaccination is an important determinant of success. 702 16-25 Bordetella pertussis vaccination illustrates the importance of the perceived safety of a vaccine. 704 16-26 Conjugate vaccines have been developed as a result of understanding how T and B cells collaborate in an immune response. 705 16-27 Peptide-based vaccines can elicit protective immunity, but they require adjuvants and must be targeted to the appropriate cells and cell compartment to be effective. 705 16-28 Adjuvants are important for enhancing the immunogenicity of vaccines, but few are approved for use in humans. 707 16-29 Protective immunity can be induced by DNA-based vaccination. 708 16-30 T he effectiveness of a vaccine can be enhanced by targeting it to sites of antigen presentation. 708 16-31 An important question is whether vaccination can be used therapeutically to control existing chronic infections. 709 Summary. 710 Summary to Chapter 16. 711 Questions. 711 General references. 712 Section references. 712 Appendix I The Immunologist's Toolbox 717 Immunization. 717 A-1 A-2 A-3 A-4 Haptens. Routes of immunization. Effects of antigen dose. Adjuvants. The detection, measurement, and characterization of antibodies and their use as research and diagnostic tools. A-5 Affinity chromatography. A-6 Radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), and competitive inhibition assay. A-7 Hemagglutination and blood typing. A-8 Precipitin reaction. A-9 Equilibrium dialysis: measurement of antibody affinity and avidity. A-10 Anti-immunoglobulin antibodies. A-11 Coombs tests and the detection of Rhesus incompatibility. A-12 Monoclonal antibodies. A-13 Phage display libraries for antibody V-region production. A-14 Microscopy and imaging. A-15 lmmunoelectron microscopy. A-16 Immunohistochemistry. A-17 lmmunoprecipitation and co-immunoprecipitation. A-18 lmmunoblotting (Western blotting). A-19 Use of antibodies in the isolation and identification of genes and their products. 718 720 720 720 723 723 723 724 725 727 727 729 730 731 732 734 735 735 737 737 Principles of innate and adaptive immunity E Isolation of lymphocytes. 739 A-20 Isolation of peripheral blood lymphocytes by Ficoii-Hypaque TM gradient. 739 A-21 Isolation of lymphocytes from tissues other than blood. 740 A-22 Flow cytometry and FACS analysis. 740 A-23 Lymphocyte isolation using antibody-coated magnetic beads. 742 A-24 Isolation of homogeneous T-cell lines. 742 Characterization of lymphocyte specificity, frequency, and function. 743 A-25 Limiting-dilution culture. 744 A-26 ELISPOT assays. 745 A-27 Identification of functional subsets ofT cells by staining for cytokines. 746 A-28 Identification ofT-cell receptor specificity using peptide:MHC tetramers. 746 A-29 Assessing the diversity of the T-cell repertoire by 'spectratyping.' 748 A-30 Biosensor assays for measuring the rates of association and disassociation of antigen receptors for their ligands. 749 A-31 Stimulation of lymphocyte proliferation by treatment with polyclonal mitogens or specific antigen. 750 A-32 Measurements of apoptosis by the TUNEL assay. 751 A-33 Assays for cytotoxic T cells. 751 A-34 Assays for CD4 T cells. 751 Detection of immunity in vivo. 753 A-35 Assessment of protective immunity. 753 A-36 Transfer of protective immunity. 753 A-37 T he tuberculin test. 754 A-38 Testing for allergic responses. 754 A-39 Assessment of immune responses and immunological competence in humans. 754 A-40 T he Arthus reaction. 756 A-41 Adoptive transfer of lymphocytes. 756 A-42 Hematopoietic stem-cell transfers. 757 A-43 In vivo depletion ofT cells. 757 A-44 In vivo depletion of B cells. 757 A-45 Transgenic mice. 758 A-46 Gene knockout by targeted disruption. 759 Appendix II CD Antigens 763 Appendix Ill Cytokines and Their Receptors 779 Appendix IV Chemokines and Their Receptors 782 Biographies 784 Photograph Acknowledgments 785 Glossary 786 Index 823
AN INTRODUCTION TO PART I IMMUNOBIOLOGY AND INNATE IMMUNITY Chapter 1 Basic Concepts in Immunology Principles of innate and adaptive immunity. The effector mechanisms of adaptive immunity. Chapter 2 Innate Immunity:The First Lines of Defense The first lines of defense The complement system and innate immunity Chapter 3 The Induced Responses of Innate Immunity Pattem recognition by cells of the innate immune system. Induced innate responses to infection
PART I AN INTRODUCTION TO IMMUNOBIOLOGY AND INNATE IMMUNITY Chapter 1 Basic Concepts in Immunology Principles of innate and adaptive immunity. The effector mechanisms of adaptive immunity. Chapter 2 Innate Immunity: The First Lines of Defense The first lines of defense. The complement system and innate immunity. Chapter 3 The Induced Responses of Innate Immunity Pattern recognition by cells of the innate immune system. Induced innate responses to infection
1 Basic Concepts in Immunology When infection does occur,how does the body eliminate the invader and cure by immunology which we study to understand our bodys defenses against infection at the cellular and molecular levels. relatively mild disease of cowpox,or vaccinia,seemed to confer protection e often ratal dlseas of smallpox. 1796,1 ner onstat ed tha e-causing aentsvgoproicdeDrotectionftom become universal,an advance that enabled the World Health Organization to When lenner introduced vaccination he knew nothing of the infectiou e disease:it was not until late in the 19th century that Rober es are ause egories of disease-causing microorganisms,or pathogens:viruses,bacteria fungi,and the unicellular and multicellular eukaryotic organisms collectively termed parasites. The discoveries of Koch and other great 19th-century microbiologists extended Jenners strategy of vaccir ation toother diseases.In the1880s,Louis g1.1 Edward r Portrait byJo el Smith. by a rabid dog.These practical triumphs led to a search for the mechanism of protection and to the development of the science of immunology.In the Whitney Medical Library
Basic Concepts in Immunology Immunology is the study of the body's defense against infection. We live surrounded by microorganisms, many of which cause disease. Yet despite this continual exposure we become ill only rarely. How does the body defend itself? When infection does occur, how does the body eliminate the invader and cure itself? And why do we develop long-lasting immunity to many infectious diseases encountered once and overcome? These are the questions addressed by immunology, which we study to understand our body's defenses against infection at the cellular and molecular levels. Immunology is a relatively new science. Its origin is usually attributed to Edward Jenner (Fig.l.1), who observed in the late 18th century that the relatively mild disease of cowpox, or vaccinia, seemed to confer protection against the often fatal disease of smallpox. In 1796, Jenner demonstrated that inoculation with cowpox could protect against smallpox. He called the procedure vaccination, and this term is still used to describe the inoculation of healthy individuals with weakened or attenuated strains of disease-causing agents to provide protection from disease. Although Jenner's bold experiment was successful, it took almost two centuries for smallpox vaccination to become universal, an advance that enabled the World Health Organization to announce in 1979 that smallpox had been eradicated (Fig. 1.2), arguably the greatest triumph of modern medicine. When Jenner introduced vaccination he knew nothing of the infectious agents that cause disease: it was not until late in the 19th century that Robert Koch proved that infectious diseases are caused by microorganisms, each one responsible for a particular disease. We now recognize four broad categories of disease-causing microorganisms, or pathogens: viruses, bacteria, fungi, and the unicellular and multicellular eukaryotic organisms collectively termed parasites. The discoveries of Koch and other great 19th-century microbiologists extended Jenner's strategy of vaccination to other diseases. In the 1880s, Louis Pasteur devised a vaccine against cholera in chickens, and developed a rabies vaccine that proved a spectacular success upon its first trial in a boy bitten by a rabid dog. These practical triumphs led to a search for the mechanism of protection and to the development of the science of immunology. In the Fig. 1.1 Edward Jenner. Portrait by John Raphael Smith. Reproduced courtesy of Yale University, Harvey Cushing/John Hay Whitney Medical Library
2 Chapter 1:Basic Concepts in Immunology early 1890s,Emil von Behring and Shibasaburo Kitasato discovered that the diphtheria or tetanus toxins in people.This activity was due to the proteins aaeoaeshhondpea6cayoaeioindim The responses we make against infection by potential pathogens are known as immune responses.A specific immune response,such as the production pve nune pheenio nath distinguish an adaptive immune response from the innate immune response. for any individual pathoge chiefly through he work d h Fig.1.2 The eradication of small ready to act,and are a front-line component of innate immune re was able against poliovirus. es co uld be induce nation al against a vast ran and some fear that these production is not the only function of adaptive immune r esponses.and the in de and s alia in and polysaccharides of pathogens are the antigens normally responded to courtesy of Dr. Jason Weistelc by the mmune e a response to a much immune responses to metals such as nickel,drugs such as penicillin.and organic chemicals in the le responses toger the pro in those that cannot be resolved.the activities of the innate immune system y lasting immunologic emory,whic This book describes the mechanisms of both innate and adaptive imm known as lymphocytes possess the most powerful nize and e system to ini e and to m mechami oly eimndimng microonn f the same hhCandada e mn
3 Chapter 1: Basic Concepts in Immunology Number of 30 countnes with one or more cases per month 15 smallpox officially eradicated o��������� 1965 1970 1975 1980 Year Fig. 1.2 The eradication of smallpox by vaccination. After a period of 3 years in which no cases of smallpox were recorded, the World Health Organization was able to announce in 1979 that smallpox had been eradicated, and vaccination stopped (upper panel). A few laboratory stocks have been retained, however, and some fear that these are a source from which the virus might reemerge. Ali Maow Maalin (lower panel) contracted and survived the last case of smallpox in Somalia in 1978. Photograph courtesy of Dr. Jason Weisfeld. early 1890s, Emil von Behring and Shibasaburo Kitasato discovered that the serum of animals immune to diphtheria or tetanus contained a specific 'antitoxic activity' that could confer short-lived protection against the effects of diphtheria or tetanus toxins in people. This activity was due to the proteins we now call antibodies, which bound specifically to the toxins and neutralized their activity. The responses we make against infection by potential pathogens are known as immune responses. A specific immune response, such as the production of antibodies against a particular pathogen or its products, is known as an adaptive immune response because it is developed during the lifetime of an individual as an adaptation to infection with that pathogen. In many cases, an adaptive immune response also results in the phenomenon known as immunological memory, which confers lifelong protective immunity to reinfection with the same pathogen. This is just one of the features that distinguish an adaptive immune response from the innate immune response, or innate immunity, which is always immediately available to combat a wide range of pathogens but does not lead to lasting immunity and is not specific for any individual pathogen. At the time that von Behring was developing serum therapy for diphtheria, innate immunity was known chiefly through the work of the great Russian immunologist Elie Metchnikoff, who discovered that many microorganisms could be engulfed and digested by phagocytic cells, which he called 'macrophages.' These cells are always present and ready to act, and are a front-line component of innate immune responses. In contrast, an adaptive immune response takes time to develop and is highly specific; antibodies against the influenza virus, for example, will not protect against poliovirus. It quickly became clear that antibodies could be induced against a vast range of substances. Such substances were called antigens because they could stimulate antibody generation. Much later, it was discovered that antibody production is not the only function of adaptive immune responses, and the term antigen is now used to describe any substance that can be recognized and responded to by the adaptive immune system. The proteins, glycoproteins, and polysaccharides of pathogens are the antigens normally responded to by the immune system, but it can recognize and make a response to a much wider range of chemical structures-hence its ability to produce allergic immune responses to metals such as nickel, drugs such as penicillin, and organic chemicals in the leaves of poison ivy. The innate and adaptive immune responses together provide a remarkably effective defense system. Many infections are handled successfully by innate immunity and cause no disease; in those that cannot be resolved, the activities of the innate immune system trigger an adaptive immune response. If the disease is overcome, the adaptive immune response is often followed by lasting immunological memory, which prevents disease if reinfection occurs. This book describes the mechanisms of both innate and adaptive immunity and illustrates how they are integrated into an effective overall system of defense against invasion by pathogens. Although the white blood cells known as lymphocytes possess the most powerful ability to recognize and target pathogenic microorganisms, they need the participation of the innate immune system to initiate and to mount their offensive. Indeed, the adaptive immune response and innate immunity use many of the same destructive mechanisms to finally destroy invading microorganisms. In this chapter we first introduce the principles of innate and adaptive immunity, the cells of the immune system, the tissues in which they develop, and the tissues through which they circulate. We then outline the specialized functions of the different types of cells and the mechanisms by which they eliminate infection
Principlesofnae and adaptive immunity 3 Principles of innate and adaptive immunity ted from infectiou cells and molecules that together make up the immune system.In this parto the chapter response depends. e sys(c Teoneesysemecognzasimectoandhdlcespoiete ses. ect the individual effectively against disease.the in detected.This task both by the white bloo and by the lymphocvtes of the adaptive immune system.The second task is to contain the infection and if possible eliminate it completely,which b into play imm s sucn as the capacities of lymphocytes and the other white blood cells.At the same time the immune respom must be kept tha it does not itsell de self-regulareis由u ant fea mune and failure of such regulation contributes to conditions such as allergy and autoimmune aseThe fourth sk Is to protect t is that it is capable of generating immunological me y,so that having once to an infectious gent,a person will m e to salarga jo auo s!ayood Ane suaoted o Alunuu! When an individual first e rs an infectiou the initial defe against infection are physical and chemical barriers,such as antimicrobia at mucosal surtaces.that prevent microbesromn immune syst m come into plav The comple ment svst em can immediately ognize and d estroy foreign orgar d phag ocytic white blood cells of the mune system ca dative enzymes.Innate immunity is of ancient origin some form of innate defense against ease is found in all animals and plants.T macrophages evolutionary dese cendants of the cytic cells p sent in sin pler animals resp ure to an infecti m take days e han hours to develop (summarized in Fig.1.34).However,the adaptive is cap f h can rec ize and respond to individual antigens by means of highly spe cialized antigen receptors on the lymphocyte surface.The billions of lym the imn
Principles of innate and adaptive immunity. The body is protected from infectious agents and the damage they cause, and from other harmful substances such as insect toxins, by a variety of effector cells and molecules that together make up the immune system. In this part of the chapter we discuss the main principles underlying immune responses and introduce the cells and tissues of the immune system on which an immune response depends. 1-1 The immune system recognizes infection and induces protective responses. To protect the individual effectively against disease, the immune system must fulfill four main tasks. The first is immunological recognition: the presence of an infection must be detected. This task is carried out both by the white blood cells of the innate immune system, which provide an immediate response, and by the lymphocytes of the adaptive immune system. The second task is to contain the infection and if possible eliminate it completely, which brings into play immune effector functions such as the complement system of blood proteins, the antibodies produced by some lymphocytes, and the destructive capacities of lymphocytes and the other white blood cells. At the same time the immune response must be kept under control so that it does not itself do damage to the body. Immune regulation, or the ability of the immune system to self-regulate, is thus an important feature of immune responses, and failure of such regulation contributes to conditions such as allergy and autoimmune disease. The fourth task is to protect the individual against recurring disease due to the same pathogen. A unique feature of the adaptive immune system is that it is capable of generating immunological memory, so that having been exposed once to an infectious agent, a person will make an immediate and stronger response against any subsequent exposure to it; that is, they will have protective immunity against it. Finding ways of generating long-lasting immunity to pathogens that do not naturally provoke it is one of the greatest challenges facing immunologists today. When an individual first encounters an infectious agent, the initial defenses against infection are physical and chemical barriers, such as antimicrobial proteins secreted at mucosal surfaces, that prevent microbes from entering the body. If these barriers are overcome or evaded, other components of the immune system come into play. The complement system can immediately recognize and destroy foreign organisms, and phagocytic white blood cells, such as macrophages and neutrophils of the innate immune system, can ingest and kill microbes by producing toxic chemicals and powerful degradative enzymes. Innate immunity is of ancient origin-some form of innate defense against disease is found in all animals and plants. The macrophages of humans and other vertebrates, for example, are presumed to be the direct evolutionary descendants of the phagocytic cells present in simpler animals, such as those that Metchnikoff observed in the invertebrate sea stars. Innate immune responses occur rapidly on exposure to an infectious organism. In contrast, responses by the adaptive immune system take days rather than hours to develop (summarized in Fig. 1.34). However, the adaptive immune system is capable of eliminating infections more efficiently because of the exquisitely specific recognition functions of lymphocytes. These cells can recognize and respond to individual antigens by means of highly specialized antigen receptors on the lymphocyte surface. The billions of lymphocytes present in the body collectively possess a vast repertoire of antigen receptors, which enables the immune system to recognize and respond to Principles of innate and adaptive immunity E