When the history of immunology in the twentieth century is written, the decade of the 1960's will, in all probability, stand out as the period of greatest advance in the development of molecular immunology. It is appropriate and useful, therefore, that a schol arly and integrated presentation of this progress should be made available in English. The translation of Dr. Nezlin's "Biochem istry of Antibodies" from Russian admirably fulfills this need in the form of a scientific monograph directed to medical and biolog ical scientists. The appearance of this monograph also serves to emphasize the conceptual unification of diverse immunological phenomena which has emerged from progress in molecular immunology. This unity is a consequence of the key role played by the antibody mol ecule (either in solution or cell-bound) in every biological process properly described as immunological. Indeed, immunology as an independent natural science can be described as the study of the structure, interactions, and biosynthes is of the antibody molecule."

Contemporary immunobiology is an ever-diversifying field that embraces many aspects of cellular and humoral immune responsiveness. This includes the phylogeny, ontogeny, induction, regulation, expression, and differentiation of the immune-cell series, as well as the relationship of these to the pathogenesis of many disease processes. It is clearly beyond the limitations of each volume in this series to comprehensively cover progress in each of these fields. Three general areas of contemporary investigations are discussed in the current issue: cellular events of B cell differentiation, including antigen recognition and respon siveness of B lymphocytes; regulation of the immune response by T cells, and the roles of this regulatory system in allotype suppression and in autoimm unity; and the genetic control of immune responses and its possible relation to disease pathogenesis. Analysis of responsiveness of B lymphocytes has been greatly aided by the availability of athymic nude mice. The immunological responsiveness of these mice to a wide range of stimuli has been reviewed and analyzed by H. H. Wortis. This article concludes with the provocative thesis that in the future "nudes will supply new rather than confirmatory data . . . (especially regarding) the develop ment of neoplasia. " In contrast to the studies on the congenitally athymic mice, G. F."

The Seventh International Conference of the Inflammation Research Association, entitled "Inflammation, Mechanisms and Therapeutics" was held on September 25-29,1994 in White Haven, Pennsylvania. The major focus of this series of conferences is the multidisciplinary investigation of inflammation and the discovery of novel therapeutic approaches to inflammatory diseases. It was therefore particularly gratifying that the list of attendees included scientists of diverse backgrounds - preclinical biology, synthetic and theoretical chemistry, biotechnology and clinical research. The Conference was characterized by a very high degree of participation in the scientific debate by attendees, a large proportion of whom presented at poster sessions, poster discussions and workshops, and by the many opportunities for informal discussion provided by the organizers. This volume captures much of the excitement and enthusiasm of the Conference and should be a valuable resource for scientists in the field. Niall S. Doherty Barry M. Weichman Douglas W. Morgan Lisa A. Marshall IX Acknowledgments The manuscripts contained in this volume are based on material presented at the Seventh International Conference of the Inflammation Research Association held in White Haven, Pennsylvania, on September 25-29,1994. We would like to thank the speakers and session chairpersons for their contributions to the Conference and these Proceedings. The following individuals played indispensable roles in the organization and running of the Conference and are responsible for making it such a success: PLANNING SCHOLARSHIP Niall S. Doherty Janet Kerr, Chairperson Barry M. Weichman Richard Carlson Douglas Morgan Richard Harris Loran Killar D.

Immunoregulation is one of the areas which has witnessed the most explosive advances of immunology during the past decade. It is in this area that the current view of the immune system has arisen and developed. There is indeed little doubt that immune reactions are primarily determined by messages which are genera ted within the immune system and passed among different types of immunologie cells. This cell communication not only determines the type, intensity and duration of the response after perturbation of the immune system by exogenous antigens, but it is also essential for preventing autoimmune reactions and their clinical conse quences. In order to assure aperfect balance within the enormous com plexity of the immune system, it is not surprising that multiple self-regulatory mechanisms are organized at different levels, such as antibody feedback, idiotypic-anti-idiotypic responses, suppres sor and helper T cells, lymphokine signals and genetic require ments. A nu mb er of observations in recent years have, however, demonstrated that consistent contributions to the immunological homeostasis are given also by signals generated outside of the immune system, namely, in the central and autonomous nervous system as weIl as in the endocrine apparatus. Furthermore, the interactions between the immune system and the other body homestatic mechanisms seem to be bidirectional: if immunological cells may be targets of neuroendocrinological factors, immunological products seem in turn to contribute to the neuro endocrine homeostasis."

The ever-expanding research on human cancer has resulted in numerous technical and conceptual advances during the last few years. Serological, structural, and biological char acterization of human melanoma constitutes one area of research that has received consid erable attention from researchers and clinicians and has generated new and exciting infor mation. In this volume, we have attempted to assemble work on topics that produced some of the most recent advances. We asked each author to describe and interpret his most cur rent research and, whenever possible, to compare and contrast it with work of other inves tigators in the field. We have been careful not to impose our viewpoints except in contri butions from our own laboratories, since we want to provide the reader with as many divergent and sometimes opposing viewpoints as feasible. Therefore, we have not been overly concerned with overlaps in some individual topics. We hope that this volume will provide the reader with a well-balanced overview of current problems and ideas in a par ticular area of cancer research. We wish to express our thanks to all contributors for their timely and very interesting manuscripts, and we sincerely hope that the reader will enjoy this volume and benefit as much from it as we did. R. A. Reisfeld S. Ferrone La Jolla ix Contents CHAPTER 1 Immunogenetics of Melanoma RONALD T. ACTON, CHARLES M. BALCH, BRUCE BUDOWLE, RODNEY C. P. Go, JEFFREY M. ROSEMAN, SENG-jAW SOONG, AND BRUCE O. BARGER 1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The phenomenon of idiotypy was discovered almost thirty years ago, but it was only during the past decade that it attracted widespread interest and became the subject of numerous research investigations. From the outset, much of the interest in idiotypy was based on its implications with respect to the repertoire of antibodies. Kunkel showed, for example, that idiotypes associated with certain human myeloma or Bence-Jones proteins were present in normal human globulins at levels of less than one part per million. Also, Oudin's original definition of idiotypy implied that idiotypes could be uniquely associated with individual rabbits as well as with particular antigen-binding specificities. Such observations provided some of the earliest evidence for an extensive repertoire of immunoglobulin molecules. The implications of these findings have been amply confirmed by recent studies of protein struc ture and molecular genetics; many of these studies are reviewed in the present volume. It is known now that the diversity of antibodies is based on the presence of numerous V and L V H genes, on recombinatorial events involving D and] segments, on somatic mutations, and on processes involving deletion of DNA followed by repair with errors, including inser tions. Each of these parameters is capable of influencing the idiotype expressed by the final immunoglobulin product. Regulation of the immune response is another area in which idiotypy has significantly influenced modern immunology.

The articles in this volume represent papers delivered by invited speakers at the 6th International Symposium on the Immunobiology of Proteins and Peptides. In addition, a few of the abstracts submitted by participants were scheduled for minisymposia and some of the authors, whose presentations were judged by the Scientific Council to be of high quality, were invited to submit papers for publication in this volume. This symposium was established in 1976 for the purpose of bringing together, once every two or three years, active investigators in the forefront of contemporary immunology, to present their findings and discuss t heir significance in the light of current concepts and to identify important new directions of investigation. The founding of the symposium was stimulated by the achievement of major breakthroughs in the understanding of the immune recognition of proteins and peptides. We believed that these breakthroughs will lead to the creation of a new generation of peptide reagents which should have enormous potential in biological, therapeutic and basic applications. This anticipated explosion has in fact since occurred and many applications of these pep tides are now being realized.

Scientific research on dengue has a long and rich history. The literature has been touched by famous names in medicine- Benjamin Rush, Walter Reed, and Albert Sabin, to name a very few- and has been fertile ground for medical historians . The advances made in those early investigations are all the more remarkable for the limited tools available at the time. The demonstration of a viral etiology for dengue fever, the recognition of mosquitoes as the vector for transmission to humans, and the existence of multiple viral variants (serotypes) with only partial cross-protection were all accomplished prior to the ability to culture and characterize the etiologic agent. Research on dengue in this period was typically driven by circumstances. Epidemics of dengue created public health crises, although these were relatively short-lived in any one location, as the population of susceptible individuals quickly shrank. Military considerations became as a major driving force for research. With the introduction of large numbers of non-immune individuals into endemic areas, dengue could cripple military readiness, taking more soldiers out of action than hostile fire. Dengue and dengue hemorrhagic fever, which assumed pandemic proportions during the latter half of the last century, have shown no indication of slowing their growth during this first decade of the twenty-first century. Challenges remain in understanding the basic mechanisms of viral replication and disease pathogenesis, in clinical management of patients, and in control of dengue viral transmission. Nevertheless, new tools and insights have led to major recent scientific advances. As the first candidate vaccines enter large-scale efficacy trials, there is reason to hope that we may soon "turn the corner" on this disease.

Interferon has been and continues to be one of the more fas cinating substances produced by apparently all animals in response to particular stimuli. It has led to major revisions in concepts of cellular immunity and theories on the recovery of multicellular systems from viral infection. Since its discovery, interferon has held the interest of the molecular biologist, and definitive answers as to its clinical value are close at hand. The following treatise is an attempt by the authors to pre sent a complete picture of the many aspects of interferon. Having recently had the privilege of visiting the laboratories of the au thors and several others of our Soviet colleagues working in the interferon field, I was most impressed with the amount of research being done in this area. Since a great deal of this work is published in the Russian language, there is unfortunately a time lag until it can be received and translated. We are therefore most grateful to Drs. Solov'ev and Bektemirov for having produced this impres sive work which offers the opportunity to review the entire field and, very importantly, some of the work being done by our Soviet counterparts, and also to Plenum Publishing Corporation for pro viding the English translation. Work on the mechanism of interferon induction, the relative role of the cellular constituents, and work in cell-free systems and the molecular mechanism of the antiviral action of interferon continues to excite scientists in the field."

When Antibiotics I was published in 1967, the teleological view was held by some that" antibiotics" were substances elaborated by certain microorgan isms for the purpose of competing with other microorganisms for survival in mixed ecological environments. However, not only had J. EHRLICH and his associates shown 15 years earlier that chloramphenicol was produced by Strepto myces venezuelae in cultures of sterilized soils but not in parallel cultures of the same soils which were not sterilized, but operationally, the search for anti cancer antibiotics was actively under way (Antibiotics I reporting on numerous such substances), although the concept of antibiosis could not logically justify such undertakings. This editor hesitates to accept the use of the term "antibiotic" for anti microbial agents of non microbiological origins which is sometimes encountered, but neither does he subscribe to the view that antibiotics are in some fundamental manner different from chemotherapeutic substances of other origins. Modes and mechanisms of action of chemotherapeutic compounds are not systematic functions of their origins nor of the taxonomical position of the target organisms. Consequently, in the selection of topics for Antibiotics III (published in 1975), synthetic drugs and natural products of higher plants (alkaloids) were represented, along with antibiotics in the strict sense of the definition. We now present Antibiotics V, for whose assembly the same selection criteria were applied as for Antibiotics Ill. The aggregate length of the contributions rendered it impractical to place the entire text between the covers of one book."

Recent advances in the understanding of the major events that shape the immune recog nition system have been remarkable. The analysis of immunoglobulin (Ig) gene organization and Ig repertoire diversification in lower vertebrates has provided new insight into this process in mammals. Similarly, the understanding of the early development of lymphocytes and of the acquisition of immunological tolerance has been aided by elegant studies in quail/chicken chimeras, using the power of the distinctive markers of the constitutive cells of these birds. Great strides have been made in understanding the role played by major histocompatibility complex (MHC) molecules in antigen presentation and in repertoire selec tion within the thymus. The use of transgenic mice expressing specific T-cell receptor (TCR) genes has elucidated the process of both positive and negative selection. In parallel, there has been considerable progress in our understanding of tolerance, based in part on the use of markers for the V fJ genes of T-cell receptors and in part on the analysis of the behavior of long term T-cell lines. This has led to the realization that both clonal deletion and clonal anergy may play critical roles in the maintenance of unresponsiveness to self antigen. Molecular analysis of the requirements for expression of membrane immunoglobulin molecules has revealed the existence of a complex that appears to be of critical importance in mediating signalling through Ig receptors. In addition, major insights have been obtained into the regulation of expression of genes of immunologic interest.

The two main goals of the symposium upon which this volume is based were 1) to cement together knowledge presently available in the field of antibodies to steroids and obtainable only under separate covers in different journals and books, and 2) to present new data which could lead to a more complete understanding of physiologic phenomena like those occurring during the menstrual cycle, or to the elucidation of the mechanisms involved in steroid-protection interaction, or to the practical application of immunologic techniques to measurements of steroid hor mones. These techniques are extremely sensitive and can measure levels of steroid on the same order of magnitude as the radioisotope methods. However, the latter are much more laborious and costly which limits their use in many cases to the research laboratory. But the immunologic techniques generally classified as radioimmunoassay, are fraught with difficulties and problems which must be overcome. Fortunately, perhaps, the subject of immunologic techniques as applied to steroid determination is the child of radioimmunoassay of proteins, so to speak. Many of the problems which confront the former have been resolved in the latter instance. Thus, we are in an advantageous position because we are aware of the biologic and technical problems of the earlier radioimmunoassay techniques. Similar experiences have been reported in the book about the use of immunologic techniques for determination of steroid hormones.

The importance of thymus-dependent cells, or T cells, in the generation of a successful immune response was first realized in the early sixties. In the follow ing two decades, a succession of elegant experiments established the antigen specificity of T cells and their ability to perform both as regulatory and effector cells. T cells were shown to be essential in most immune reactions, playing a crucial role in augmenting the activity of effector T and B cells against 'foreign' antigen, as well as in the suppression of effector activity against self antigens. The means by which T cells differentiate 'foreign' from 'self' antigens is based on their recognition of antigen almost exclusively in the context of self major histocompatibility complex products, unlike B cells, which recognize an tigen alone. It is this recognition, mediated by the T-cell receptor, that sets into motion the diverse cell-cell interactions, which control the differentiation and regulation of the immune response. Although its importance was well established, the molecular nature of the T-cell receptor remained elusive for two decades. Many hypotheses as to its structure and precise function were put forward, using immunoglobulin as a basis for conjecture, but "the Holy Grail of Immunology" remained ephemeral until three years ago. In the ensuing years, both immunologists and molecular biologists have contributed to an explosion of data unsurpassed by any previous period in the field."

Since the discovery more than thirty years ago that antibody actlvlty could be localized to discrete plasma protein fractions, the study of immunoglobulin struc ture and function has dominated the field of immunochemistry. During this time, sources of homogeneous immunoglobulin molecules have been discovered, the subunit nature of the proteins has been defined, and the three-dimensional struc tures of the antigen-recognition portion of several antibody molecules have been elucidated. Insights into the complicated genetic control of these proteins are being gained rapidly through analysis of amino acid sequences of naturally occurring and induced homogeneous immunoglobulins. Immunoglobulins have been analyzed by protein chemists as models of complex multimeric systems, examined by geneticists studying serum protein polymorphisms, and employed by molecular biologists as highly selective probes capable of distinguishing minor features of molecular topog raphy. Clinical applications have ranged from the now routine quantitation of immunoglobulin levels to the use of antibodies to detect trace levels of a variety of natural products and drug metabolites. All these applications have depended ulti mately on a thorough understanding of the immunoglobulin and its antigen-combin ing site. To cover the entire field of immunoglobulin structure and function would require many volumes this size; therefore, subjects presented in this volume represent those which we felt contribute most to our current understanding of this protein family. The first chapters deal with the structure and function of the immunoglobulin molecule."

This volume contains the main papers presented at the 1997 EUROTOX Congress, Arhus, Denmark, 24-28 June 1997. Diversification in toxicology is seen as the application of basic science to such diverse areas as man and his environment. The pressing issues which have been dealt with not only include reproductive effects of environmental chemicals ("xenoestrogens"), but also receptor-mediated toxic responses, new frontiers in human and ecological toxicology, chemoprevention of cancer and molecular approaches in toxicological research. The practical and ethical facets of toxicology, e.g. ecotoxicological risk assessment, biomarkers of exposure, complex chemical mixtures as well as animal welfare and the ethics of animal experimentation, are also treated.

The Antibody Enigma is a somewhat personal view of the antibody diversity question from two investigators who have spent the past 18 years trying to penetrate the enigma. It is not and was not meant to be an all-embracing comprehensively referenced review of the subject of antibody diversity. Because of the subjective viewpoint, there are un doubtedly omissions of data that others consider to be seminal, and if we have offended anyone by omitting their own contribution we sin cerelyapologize. We have lived with "The Enigma" on and off for the past two years. It has been both hard work and good fun but, above all, it has been a learning experience. There were several difficult decisions to make in putting together the final text, but perhaps the most difficult was de ciding upon a stopping point. The field of antibody diversity is presently enjoying an unparalleled expansion of information, and because of this it was very tempting to await further developments in hopes of tying up as many loose ends as possible. This was decided against for several reasons; the major factor was that the project was growing burdensome for both of us. From a more objective point of view this appears to be a reasonable time to stop our exposition."

Our understanding of the function of natural killer (NK) cells has dramatically changed in recent years. The discovery of NK receptors specific for MHC class I molecules, and the study of the role of co-stimulatory and adhesion molecules have led to an understanding of how NK cells recognize tumor and virally infected cells that have lost expression of MHC class I molecules or have altered distribution of normal cell surface molecules. Such recognition events lead to intracellular signals which can be either stimulatory or inhibitory. This book provides an insight into how NK cells develop, how they learn to distinguish altered cells from normal cells, and into their biological role in controlling infections and tumors.

In this current volume of Contemporary Topics in Immunobiology we have chosen to continue with the multiple-theme approach that was developed in Volumes 1, 3, and 5 of this series. Immunobiology still shows little sign of decreasing its active growth rate, but rather is continuing to broaden its range of interests and applications, particularly as new techniques and methods are adapted from other fields of medical research. This present volume reflects both several of the more classical areas of immunology now addressed in the light of contemporary immunology, and several newer directions that have been taken in other fields. The general subject of T-cell heterogeneity and functions of T-cell subpop ulations is addressed in Chapters 1 and 2. The potential role of genes of the major histocompatibility complex in controlling the immune functions of T lymphocytes still remains a major unresolved issue in immunogenetics, and the current status of this problem is excellently reviewed by J. F. A. P. Miller. The further elucidation of functional subpopulations of human T lymphocytes has been particularly hampered by the lack of available markers for character izing and isolating such sUbpopulations. A major step in this direction has been made by L. Moretta, M. Ferrarini, and M. D. Cooper, who review their ex perience with Fc-receptor-bearing human T-Iymphocyte populations."

There has been a tremendous increase in interest in the neuropathogenicity of viruses during the past decade as we have come to recognize that the human immunodeficiency virus, which causes the acquired immunodeficiency syndrome (AIDS), can infect glial cells and cause neurological disease. Yet this increase has not been limited to AIDS but has extended to viruses that infect either or both the central and peripheral nervous systems. The changes examined here include both neurological and psychological diseases or syndromes. Moreover, the chapters in this volume review the interaction of the host immune system with the viruses examined and how such interactions may increase or decrease the neuropatho genicity of the viruses. Questions regarding viral neuropathogenesis include: (I) What is the mode of transmission of virus to the nervous system? (2) What types of cells are infected, and do they contain receptors for the virus? (3) What is the extent of damage that results from viral infection? (4) What are the immunologic mecha nisms by which damage is mediated or limited? Many of these questions remain unanswered, but this volume delves into efforts to provide some answers.

Ever since arbovirus infections became known and their relative importance assessed, experiments were designed to elucidate the mode of transmission and the most important natural hosts responsible for perpetuating the infection in nature. Human infections and the disease in wild rodents, birds, and domestic animals were studied in relation to viremia and distribution of the infectious agent in the organism. With increasing epidemiological studies it became apparent that the neural manifestations of the disease are very uncommon, confined only to a small percentage of individuals of the most susceptible species. Various factors have been proposed to explain why in certain instances the virus becomes establish ed in the central nervous system and causes a serious or lethal disease. For example, differences in the virulence of the virus strains, varying susceptibility of individuals of one species, or intercurrent circumstances facilitating access of the virus to the central nervous system were alleged. Also, various possible routes of entry of the virus into the brain and spinal cord have been considered."

The proper physiological functioning of most eukaryotic cells requires their assembly into multi-cellular tissues that form organized organ systems. Cells of the immune system develop in bone marrow and lymphoid organs, but as the cells mature they leave these organs and circulate as single cells. Antigen receptors (TCRs) of T cells search for membrane MHC proteins that are bound to peptides derived from infectious pathogens or cellular transformations. The detection of such speci?c peptide-MHC antigens initiates T cell activation, adhesion, and immune-effectors functions. Studies of normal and transformed T cell lines and of T cells from transgenic mice led to comprehensive understanding of the mole- lar basis of antigen-receptor recognition and signaling. In spite of these remarkable genetic and biochemical advances, other key physiological mechanisms that par- cipate in sensing and decoding the immune context to induce the appropriate cellular immune responses remain unresolved. TCR recognition is tightly regulated to trigger sensitive but balanced T cell responses that result in the effective elimination of the pathogens while minimizing collateral damage to the host. The sensitivity of TCR recognition has to be properly tempered to prevent unintended activation by self-peptide-MHC complexes that cause autoimmune diseases. It is likely that once the TCR is engaged by a peptide- MHC and TCR signaling begins, additional regulatory mechanisms, involving other receptors, would increase the ?delity of the response.

It was not too long ago that many physicians and biomedical scientists felt that the era of 'vaccines' for protecting mankind against infectious disease was coming to an end. During the 1 940s and 50s the widespread use of newly developed antibiotics and antimicrobial chemotherapeutic agents suggested a new era in medicine, i. e. the control and eventual elimination of all infectious diseases, at least those caused by bacteria, by' chemical means. The magic 'bullet' proposed by Paul Ehrlich in the early 1900s seemed to be the method of choice for controling infection. However, it is now quite evident that those high expectations were unwarranted. Although many acute infections, especially those caused by pyogenic cocci, have been controlled by antibiotics, it is quite evident that infectious diseases, even those caused by bacteria, still are a major problem. Thus, the old 'standby' of preventative vaccination is making a strong comeback, not only for viral but also for bacterial infections. However, except for a relatively small number of viral diseases and those bacterial diseases due to toxin elaborated by microorganisms rather than invasion and replication of the microbe per se, preventative vaccination still has not fulfilled the expectations of their proponents. There has been a recent resurgence of interest concerning all aspects of vaccines, not only their preparation and administration, but also the nature and mechanism of the host immune response to the constituent micro organisms and their products."

"When we give a definition it is for the purpose of using it." HENRI POINCARE in Science and Method A. Objectives The first version of this paper was written to introduce new students and fellows of my laboratory to the mysteries of herpesviruses. Consonant with this design sections dealing with well documented data were trimmed to the bone whereas many obscure phenomena, controversial data and seemingly trivial observations were discussed generously and at length. There is some doubt as to whether it was meant to be published, but it was not a review. The objective of reviews is frequently to bring order. But alas, even the most fluent summation of credible data frequently makes dull reading and too much plausible order, like very little entropy in chemical reactions, is not the most suitable environment on which to nurture the urge to discover. This version is more charitable but not less inbalanced. The bibliography reflects the intent of the paper and was updated last in December of 1968. It should be obvious without saying that no single account such as this can do justice or injustice, as the case may be, to the several hundred papers published on herpesviruses each year or to the many thousand papers published on herpesviruses since the first of the members of the family was experimentally transmitted to a heterologous host more than half a century ago (GRUTER, 1924). B. Definition 1.