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The biological activity of soluble antigen-antibody complexes. II. Physical properties of soluble complexes having skin-irritating activity

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Abstract

Soluble BSA-anti-BSA complexes, formed in antigen excess, give immediate skin reactions in normal guinea pigs. The mechanism of the reaction is not that of passive or reversed passive anaphylaxis. The complex itself is toxic. Skin activity of the complex depends on its composition. It has become obvious that the complex composed of two antigen molecules and one antibody molecule, (Ag(2)Ab), does not have the activity, whereas, Ag(3)Ab(2) and more complicated complexes do. The role of complement as well as speculation on the structural changes of antibody-antigen complexes is presented.

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... Another factor may relate to the varied degree of complexity of soluble complexes formed. There is abundant evidence relating biological properties of immune complexes to their molecular size (10)(11)(12). It was of interest that one animal (16-23), that had predominately a low molecular weight complex in vivo, exhibited minimal inflammatory lesions--yet had chronic proteinuria with mainly membranous changes. ...
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Three of 16 rabbits injected (intravenously) daily with crystalline bovine serum albumin (BSA) for periods in excess of 10 wk developed chronic glomerulonephritis. In vivo, animals with chronic proteinuria formed variable quantities of soluble complex after injection of antigen while animals without proteinuria exhibited rapid removal of the injected BSA. In vitro studies demonstrated that a major part of the antibodies produced by rabbits with chronic nephritis lacked precipitating properties. Interpretations of these observations were presented in the discussion. It is suggested that, in addition to quantity, quality of antibody plays an important role in the development of chronic serum sickness. Complexes formed with nonprecipitating antibody, which are less rapidly removed from circulation, would have a greater opportunity to deposit in glomeruli and induce inflammation.
... Subvisible particle analyses revealed major differences between the heat-aggregated mAb 36-71 preparation, which had high concentrations of micron-sized particles, and IC mAb 36-71, which had high concentrations of nanometer-sized particles. We deliberately generated IC in antigen excess to favor smaller complexes based upon literature suggesting that smaller immune complexes are more inflammatory [84,85]. Intriguingly, proliferation of CA30 T cells in the group immunized with heataggregated mAb 36-71 was somewhat diminished by pretreatment with the isotype control IgG. ...
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A paradox in monoclonal antibody (mAb) therapy is that despite the well-documented tolerogenic properties of deaggregated IgG, most therapeutic IgG mAb induce anti-mAb responses. To analyze CD4 T cell reactions against IgG in various physical states, we developed an adoptive transfer model using CD4+ T cells specific for a Vκ region-derived peptide in the hapten-specific IgG mAb 36–71. We found that heat-aggregated or immune complexes (IC) of mAb 36–71 elicited anti-idiotypic (anti-Id) antibodies, while the deaggregated form was tolerogenic. All 3 forms of mAb 36–71 induced proliferation of cognate CD4+ T cells, but the aggregated and immune complex forms drove more division cycles and induced T follicular helper cells (TFH) development more effectively than did the deaggregated form. These responses occurred despite no adjuvant and no or only trace levels of endotoxin in the preparations. Physical analyses revealed large differences in micron- and nanometer-sized particles between the aggregated and IC forms. These differences may be functionally relevant, as CD4+ T cell proliferation to aggregated, but not IC mAb 36–71, was nearly ablated upon peritoneal injection of B cell-depleting antibody. Our results imply that, in addition to denatured aggregates, immune complexes formed in vivo between therapeutic mAb and their intended targets can be immunogenic.
... Another factor may relate to the varied degree of complexity of soluble complexes formed. There is abundant evidence relating biological properties of immune complexes to their molecular size (10)(11)(12). It was of interest that one animal (16-23), that had predominately a low molecular weight complex in vivo, exhibited minimal inflammatory lesions--yet had chronic proteinuria with mainly membranous changes. ...
Article
Full-text available
Three of 16 rabbits injected (intravenously) daily with crystalline bovine serum albumin (BSA) for periods in excess of 10 wk developed chronic glomerulonephritis. In vivo, animals with chronic proteinuria formed variable quantities of soluble complex after injection of antigen while animals without proteinuria exhibited rapid removal of the injected BSA. In vitro studies demonstrated that a major part of the antibodies produced by rabbits with chronic nephritis lacked precipitating properties. Interpretations of these observations were presented in the discussion. It is suggested that, in addition to quantity, quality of antibody plays an important role in the development of chronic serum sickness. Complexes formed with nonprecipitating antibody, which are less rapidly removed from circulation, would have a greater opportunity to deposit in glomeruli and induce inflammation.
... Antigen-antibody complexes are protein macromolecules with a molecular weight of 300,000 and greater (27). The above data indicate that these complexes are cleared at about the same rate as colloidal suspensions. ...
Article
Blockade of the RES was accomplished by the intravenous injection of carbon particles, thorotrast, zymosan, or a suspension of Bordetella pertussis. The blockade resulted in a decrease in sensitivity to anaphylaxis produced by the intravenous injection of soluble antigen-antibody complexes consisting of an optimal shocking mixture of bovine plasma albumin and mouse antibody to this antigen. The decrease in sensitivity to anaphylaxis was dependent on the dose of blockading agent and on the time between blockade and challenge with complex. The loss of sensitivity to anaphylaxis could not be restored by the administration of fresh serum from normal mice nor by guinea pig complement. Antigen-antibody complexes were rapidly removed from the blood with an average half-time of 11.9 minutes in normal mice. Complexes were cleared at significantly more rapid rates in mice previously sensitized to antigen. Although not all the results can be explained on the basis of blockade the facts indicate that the RES does play an important but as yet undefined role in passive homologous anaphylaxis in the mouse.
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In recent years, accumulating evidence has indicated that platelets continuously repair vascular damage at sites of inflammation and/or infection. Studies in mouse models of inflammation have highlighted the fact that the mechanisms underlying bleeding prevention by platelets in inflamed organs can substantially differ from those supporting primary hemostasis following tail tip transection or thrombus formation in models of thrombosis. As a consequence, exploration of the hemostatic function of platelets in inflammation, as well as assessment of the risk of inflammation-induced bleeding associated with a platelet deficit and/or the use of anti-thrombotic drugs, require the use of dedicated experimental models. In the present review, we present the pros and cons of the cutaneous reversed passive Arthus reaction, a model of inflammation which has been instrumental in studying how inflammation causes vascular injury and how platelets continuously intervene to repair it. The limitations and common issues encountered when working with mouse models of inflammation for investigating platelet functions in inflammation are also discussed.
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The results of our previous investigation indicated that the reaction of intraperitoneal cells against foreign materials depended on the heterogenicity and the molecular weight of the substance (Takeuchi, 1973). Substances tested for cellular reaction in this agar-bullet method were (1) bovine serum albumin, (2) soluble antigen-antibody complexes, (3) mixtures of bovine serum albumin and rabbit gamma globulin, (4) precipitated complexes, and (5) bovine gamma globulin. The soluble complex prepared with excess antigen in vitro showed a marked cellular reaction, which was considered to be due to its heterogenicity and molecular weight.
Of the numerous in vitro methods available for estimating the concentration of antigen or antibody in biological fluids those based on the determination of the primary interaction between antigen and antibody have certain advantages over methods in which secondary interactions are measured. The chief advantages are that (1) meaningful thermodynamic and stoichiometric characterization of the reaction can be obtained when antibody and hapten are employed, and (2) nanogram amounts of antigen or antibody can be measured relatively simply and rapidly if the antigen is radioactively labeled.
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This chapter discusses the immunology of human chrionic gonadotropin (HCG). It also reviews the immunology of HCG, examples of various disciplines involved in hormone research where immunological methods are employed. These include the characterization of HCG preparations, the detection of contaminating proteins, quantitative determination of HCG, and differentiation of HCG from other protein hormones. The relative purity of urinary extracts containing HCG is expressed as a relation between biological activity and weight (IU/mg). Employment of physicochemical and particularly immunochemical methods enable the characterization of the protein components of the urinary extracts. They serve as analytical tools to follow the efficiency of various purification procedures. Immunological methods in combination with the use of isotope-labeled HCG are introduced to the study of metabolism and fate of HCG. The possibilities for using antisera for the purification of other gonadotropic hormones and in metabolic studies are also discussed. The extraction and purification of HCG together with some of its biological and physicochemical characteristics are also described.
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Abstract1.Membranous glomerulonephritis, a major cause of the nephrotic syndrome in adults, has a typical morphologic picture with a defined initiation, evolution, progression, and reparative phase. No other renal disease evolves into it, nor does it become transformed into another type of glomerulonephritis.2.In most patients the disease appears indolent and the decline in renal function may be very slow. In some patients the course is fairly rapid, survival from the onset of illness ranging from months to several years. A few patients have had either prolonged remissions or complete recovery. Almost all recent studies conclude that steroids have no effect on proteinuria, at least during short term observation. The highly variable course of the proteinuria and the spontaneous remissions of some patients make the evaluation of any therapy difficult.3.In a small number of cases distinct conditions are associated with the occurrence of membranous glomerulonephritis. These include several types of infectious disease, gold and mercury therapy, and renal vein thrombosis.4.Current concepts of the pathogenesis postulate an unknown antigenic stimulus with an antibodyresponse appropriate to produce circulating soluble immune complexes that are deposited in the glomerulus. These deposits have been shown almost invariably to have IgG and B1c components. Their structure is such that they evidently migrate through the glomerular basement membrane, thereby possibly isolating themselves from all cells except for visceral epithelium. Such isolation may explain the lack of cellular response.
Article
The effect of addition of antigens to cultures of lymphocytes from atopic individuals was investigated. The number of large “blast cells” in cultures of patients sensitive to ragweed, Alternaria, and penicillin was greater than in control cultures from non-sensitive individuals, supporting the specific stimulatory effect of antigen. For each antigen, there is an optimal dose-response curve. Above this optimal concentration no further stimulatory effect is obtained. When two or more antigens to which a patient is sensitive were added to his lymphocytes, no cumulative stimulatory effect was obtained, as compared with the effect of only one antigen. A similar absence of summation could be seen when an antigen was mixed with phytohemagglutinin. Although the morphology of the blast cells obtained with antigenic stimulation was similar or identical to that obtained with phytohemagglutinin (PHA), the percentage of blast cells was strikingly higher with PHA, and the peak of response appeared after 3 days with PHA instead of 5 to 7 days as with antigens. When the percentage of blast cells obtained with ragweed and Alternaria was compared with other tests of hypersensitivity, such as, skin test and passive hemagglutination, a fairly good correlation was observed. On the other hand, penicillin-sensitive patients can exhibit a positive hemagglutination test and an increased percentage of blast cells with a negative skin test. When antigen was added to the washed lymphocytes of a patient sensitive to that particular antigen, the culture fluid contained antibodies demonstrable by the Schultz-Dale technique. The highest content was observed from the third to the fifth day with a significant decrease at the seventh and ninth days. Heating this culture fluid at 56° C. for 4 hours significantly decreased the content in antibodies. The addition of fluid from lymphocytes grown in tissue culture in the presence of antigen caused contraction of unsensitized monkey ileum strip. This conceivably could be due to the presence of antigen-antibody complexes.
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Microcirculatory disturbance produced on normal guinea pig mesentery by soluble IgG complexes was observed by in vivo microscopy for one hour. Soluble IgG complexes dropped onto normal guinea pig mesentery produced acute local vascular reactions which were characterized by plasma exudation, stasis, sticking and emigration of leukocytes and hemorrhage. These changes were recognized only in the venules. Evidences have been presented to show that these changes were chiefly brought about by the plasma exudation from the venules. Analyzing various phases of immunologic vascular reactions, the importance of the flow dynamic changes of microcirculatory region was emphasized.
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Fc-receptor-mediated clearance and nonspecific phagocytic clearance were assessed after the infusion of monomeric human IgG, heat-aggregated human IgG, and a monoclonal anti-mouse macrophage FcII receptor antibody (2.4G2) into normal mice. Each agent blocked Fc-receptor function in vivo, but 2.4G2 was much more potent per microgram than the other agents. Monomeric IgG in blocking doses did not affect other aspects of immune function. In contrast, aggregated IgG, and to a lesser extent, 2.4G2 reduced serum complement levels. In addition, these agents also caused moderate reductions in nonspecific phagocytic function. Monoclonal anti-mouse macrophage C3bi receptor antibody (Mac-1), another monoclonal antibody which binds to macrophage CR3 receptors without interfering with Fc-receptor function, also reduced serum complement and inhibited nonspecific phagocytic function. Complement depletion alone (produced by infusion of cobra venom factor) could not account for the observed changes in Fc receptor or nonspecific phagocytic function. We conclude that both monomeric IgG and anti-Fc-receptor antibodies can markedly inhibit Fc-receptor function in vivo; however, the pattern of physiologic changes produced by these agents differs.
Article
The Schultz-Dale technique employing in vitro passively sensitized guinea pig ileum has been used to establish quantitative relationships between antigen and antibody in anaphylaxis. To study such relationships in a laboratory model of allergy, human smooth muscle strips (ileum and uterus) were passively sensitized with serum from untreated allergic patients. After repeated washing, such strips were mounted in an isotonic Schultz-Dale apparatus and challenged with varying doses of allergen. Contractions of the strips were compared with those induced by measured doses of histamine. The responses were specific since they occurred only with the allergen. Several nonsensitized ileal and uterine strips were exposed to mixtures containing a constant amount of allergic serum and varying doses of allergen. Such strips contracted best with mixtures containing moderate amounts of allergen.Since there appears to be a parallel between these experiments and anaphylaxis induced by complexes, it is suggested that perhaps a mechanism involving complexes, similar to that proposed by Germuth, Ishizaka, and others, for anaphylaxis, is operating in allergy.
Article
Wheal-and-erythema responses were studied in normal human volunteers and in a single human subject who is sensitive to the 2,4-dinitrophenyl group. In the normal subjects, reactive skin sites were established by intradermal injection of purified rabbit antibody specific for the 2,4-dinitrophenyl group. In both the active and passively sensitized subjects, wheal-and-erythema was elicited by intradermal injection of a 2,4-dinitrophenyl protein, but not by injection of the same conjugate mixed with certain low molecular weight 2,4-dinitrophenyl haptens or with univalent fragments split by papain from anti-2,4-dinitrophenyl antibody. The latter fragments, unlike intact, bivalent, antibody, do not sensitize normal human skin sites. From these and other observations it is concluded that the wheal-and-erythema response in human skin requires mutually multivalent antigen and antibody. This requirement suggests that multimolecular complexes, containing at least 2 antigen and 2 antibody molecules, are essential in the pathogenesis of this allergic response.
Article
Blockade of the RES was accomplished by the intravenous injection of carbon particles, thorotrast, zymosan, or a suspension of Bordetella pertussis. The blockade resulted in a decrease in sensitivity to anaphylaxis produced by the intravenous injection of soluble antigen-antibody complexes consisting of an optimal shocking mixture of bovine plasma albumin and mouse antibody to this antigen. The decrease in sensitivity to anaphylaxis was dependent on the dose of blockading agent and on the time between blockade and challenge with complex. The loss of sensitivity to anaphylaxis could not be restored by the administration of fresh serum from normal mice nor by guinea pig complement. Antigen-antibody complexes were rapidly removed from the blood with an average half-time of 11.9 minutes in normal mice. Complexes were cleared at significantly more rapid rates in mice previously sensitized to antigen. Although not all the results can be explained on the basis of blockade the facts indicate that the RES does play an important but as yet undefined role in passive homologous anaphylaxis in the mouse.
Article
The concept of elicitation of reactions of anaphylactic type by non-tissue-fixing antibody, through activation of complement and release of anaphylatoxins by antigen-antibody complexes in vivo, is not clearly defined by published evidence. Experimental data are presented to demonstrate that guinea pig immunoglobulin G2 (noncytotropic) complexed with antigen in vitro elicits dermal reactions in guinea pigs, and that pretreatment of animals with complement-inactivating cobra venom factor diminishes such reactions. The various pathways through which immediate hypersensitive reactions may occur are discussed.
Article
The evidence for elicitation by physically and functionally univalent haptens is reviewed, and the importance of such reactions in nature is discussed. Data indicate that configuration of the hapten and binding site characteristics such as ‘depth’ may be decisive in determining effectiveness of elicitation. The probability of a ‘toxic’ configurational change of cell-fixed antibody molecules on interaction with antigen and possible differences in the mechanism of anaphylaxis produced by univalent and multivalent antigens are discussed.
Article
Comparisons of the response of monkey and guinea pig ileum and human appendix to histamine, bradykinin, serotonin, and SRS-A (slow-reacting substance) are reported. The monkey and guinea pig ileum reacted equally to both histamine and bradykinin. The guinea pig ileum showed a greater response to serotonin than did the other tissues. On the other hand, the monkey ileum reacted more strongly to SRS-A substance than did the guinea pig ileum. The human appendix responded well to both histamine and bradykinin but gave only weak contractions with serotonin. The monkey ileum and human appendix were able to be passively sensitized in vitro by much less serum of allergic patients than was required to sensitize the guinea pig ileum. This sensitization of the guinea pig ileum was not destroyed when the atopic serum was heated at 56° C. for 4 hours. However, this same treatment of serum prevented sensitization of the monkey ileum and human appendix. In vitro sensitization was enhanced 2 to 10 times when serum was diluted in isotonic buffered glucose rather than Tyrode's solution, and also if succinic or maleic acids were added with the antigen for challenge.
Chapter
This review is intended as a general survey of some basic contributions to the field of allergy and immunology particularly from the point of view of immunochemical principles, with certain aspects treated in detail for illustration. The literature cited is chiefly from English and American sources, in part because it is difficult enough to be conversant with articles in a rapidly expanding area limited by a single language. Many highly significant contributions have been omitted because of this as well as limitations of space and time.
Chapter
Jedes höher differenzierte Lebewesen hat während seines eigenständigen Lebens eine fast kontinuierliche Auseinandersetzung um die Erhaltung seiner Eigenart zu führen. Jede dem Bauplan der individuellen Eiweißstruktur fremde Substanz, welche in den Organismus — außerhalb des Verdauungstraktes — gelangt, muß den Eigengesetzlichkeiten des Empfängerorganismus unterworfen oder eliminiert werden, wenn die individuelle Existenz gesichert bleiben soll. Die somatisch-funktionellen Eigengesetzlichkeiten sind in der Eiweißstruktur eines Lebewesens genetisch verankert (s. Schriefers, Stave, Bd. IV).
Chapter
Pathogenetische Probleme hatten von jeher für die Nierenerkrankungen eine praktisch wichtige Bedeutung. Der beste Beweis ist die schnelle Anerkennung, die das „Pathogenetische System der Brightschen Nierenkrankheiten“ von Volhard und Fahr (1913) schon bald nach seiner Aufstellung gefunden hat. Erst nach dieser Klassifizierung der Nierenkrankheiten in degenerative, entzündliche und arteriosklerotische Leiden war es möglich, feste morphologische Begriffe mit den funktionellen Symptomen der Klinik zu verbinden. Prognostische Aussagen wurden möglich.
Chapter
The most exciting change in cell biology over recent decades has been the start made in defining the molecular changes that account for cellular phenomena. This progress is providing us with a firmer comprehension of the degree of complexity of living systems—an appreciation that was simply not possible as long as one was limited to studying global phenomena. If I had to pick a single point in time where molecular analysis began in immunology I would select 1939 when Elvin Kabat published the characterization of antibodies.1 It took another 25 years to identify the principal antibody (IgE) that mediates allergic responses2 and another 20 years after that until one had at least the outline of the cell surface receptor (FcεRI) through which IgE acts and that serves as the focus of this text.3
Chapter
The name lupus erythematosus (LE) covers two forms—the discoid chronic (cutaneous) form (DLE) and the systemic one (SLE). The relation between the two is unclear, because the clinical picture and course and the concomitant hematological and immunological abnormalities all display marked differences.
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Die nähere Erforschung des immunologischen Adjuvanseffektes und seine ausgedehnte Verwendung für die Immunisierungspraxis setzte nach den grundlegenden Veröffentlichungen von Ramon (1925a—c), Ramon und Descombey (1925a,b) und Glenny und ihren Mitarbeitern (1926) ein. Den Anstoß zu Ramons Untersuchungen hatten einige Beobachtungen an Pferden, die zur Antitoxingewinnung immunisiert worden waren, gegeben. Derartige Pferde hatten, wie es schien, überzufällig häufig, dannbesonders hohe Antitoxinkonzentrationen erreicht, wennam Applikationsort der Antigene lokale entzündliche Veränderungen auf getreten waren. Um diesen Zusammenhang zu prüfen, suchte Ramon nach Substanzen, die er Toxoiden (Anatoxinen) beimischen konnte und die sowohl die begrenzten lokalen entzündlichen Veränderungen als auch eine Steigerung der Antitoxinkonzentration im Blut bewirkten. Von den verschiedenen untersuchten Stoffen erwies sich ihm pulverisiertes, sterilisiertes Tapioca als besonders geeignet. Sowohl bei der Gewinnung des Diphtherieantitoxins als auch des Tetanusantitoxins wurden bei Verwendung von Tapioca Antitoxinkonzentrationen im Blut der Pferde erreicht, die um ein Vielfaches höher lagen als jene Titer, die durch die gleichen Mengen Anatoxin allein zu erzielen waren. Sehr bald erwies sich dann Tapioca auch bei anderen Antigenen als wirksames Adjuvans (Ramon et al. 1937, 1938, 1939).
Article
As the main characteristics of anaphylaxis in most species of animals and of human allergy involve reactions of smooth muscle, increased capillary permeability (edema formation) and hemodynamic changes with a predominance of fall in arterial blood pressure, it was natural to look for a principle present in the body having such pharmacodynamic actions. Among the endogenous principles known to physiologists in the earliest days of anaphylaxis, only histamine would fit the prerequisites as a mediator of the anaphylactic reactions. However as the complex picture of anaphylaxis emerged from the many-sided studies on its mechanism, it became more and more clear that histamine could not explain all symptoms which are typical of anaphylaxis. In 1941, Dragstedt defined anaphylaxis as a phenomenon of “auto-intoxication by physiologically active substances.” Therefore, besides histamine, many other endogenous principles might participate in the phenomenon, and at least one more principle, heparin, was shown to be released in dog’s anaphylaxis, explaining the decreased coagulability of the blood which is a typical occurrence in this species of animals. Today, though histamine still is the most important mediator of the anaphylactic reaction in some species, as the guinea pig and the dog, we have to look for newly identified principles such as 5-HT and bradykinin (or a slowly reacting principle) in the genesis of anaphylaxis. Other endogenous principles, as acetylcholine, catechol amines or adenylic compounds, could play a rather subsidiary or questionable role in the anaphylactic reaction in all species of animals or in human allergy.
Chapter
This chapter focuses on the fate of antigen-antibody complexes in animals, which depends on both the nature of the antigen involved and the immune state of the animal. The chapter deals only with complexes composed of soluble serum protein antigens and their specific antibodies, because a correlation between the in vivo behavior of complexes and their biological activity can be made only with these systems. The chapter examines the biological activity of antigen-antibody complexes and describes the effect of complement on the behavior of antigen-antibody complexes. The in vitro reaction of complement with antigen-antibody complexes results in a number of secondary phenomena such as lysis of sensitized erythrocytes, activation of proenzymes, production of anaphylatoxin, and precipitation of soluble antigen-antibody complexes. The role of complement in the biological reactions mediated by soluble antigen-antibody complexes is evaluated. It is shown that numerous pathological reactions occur as a result of the interaction between antigen and antibody both in vivo and in isolated tissue. The chapter demonstrates that certain soluble antigen-antibody complexes prepared with excess antigen produce anaphylaxis, smooth muscle contraction, lesions of serum sickness, and cutaneous reactions.
Article
A mouse model for the “sudden death” and “malarial lung” syndromes is described. Mice of the C3H/z strain succumb suddenly approximately 7 days after an infection with Plasmodium berghei becomes patent, at a time when parasitemia is still moderate (6 to 8%). Death could be shown to be due to anaphylactoid shock, probably induced by soluble immune complexes. Increased vascular permeability caused transudation and leakage of serum proteins into the interstitium and the alveoli. The lungs were found to be edematous, with a fine granular precipitate in the alveoli and adherent to the vascular walls. The precipitates reacted with antiglobulins G and M, and could be shown to also contain malaria antigens and . A dramatic drop in hematocrit was recorded several hours before death, indicating the sudden release of malaria antigens. The myocardium of animals that had died very suddenly showed a patchy loss of phosphorylase activity. This loss of activity was much more extensive, and sometimes almost total, when there had been an agonal period of several (1 to 3) hours before death. In these cases the irreversibility of the myocardial damage was also indicated by the loss of activity of the dehydrogenases, as well as by typical inflammatory reactions of granulocytic and histiocytic infiltrations. The hearts thus presented a typical picture of the acute and peracute shock syndromes. In acute shock cardiac insufficiency develops so suddenly that death ensues before irreversible damage has occurred, and cardiac insufficiency can only be demonstrated by the most sensitive of enzyme histochemical means. In the present case shock was induced by the anaphylactoid activity of immune complexes with the lung as target organ. The described syndrome appears analogous to human “malarial lung.”
Article
It has been a little more than 50 years since we discovered IgE, a key molecule for the allergic response and a target for treating allergies and severe asthma. Here, I trace my career, from the kindling of my interest in immunochemistry to groundbreaking discoveries in the biology and chemistry of immunoglobulins. I describe my service to the broader community of immunologists and my role in shaping departments and research institutes. My course starts in Japan and includes Southern California, Baltimore, and Denver.
Article
The sensitization process can be separated from the process of antibody uptake. The amount of antibody retained by in vitro sensitized tissues is proportional to the concentration of antibody present in the bulk phase of the solution used for sensitization. Temperature has little effect on the amount of antibody retained by the tissue as a whole but does affect the degree of sensitization achieved. The velocity of sensitization is proportional to the bulk phase concentration of antibody used. Sensitization seems to be associated with the attachment of antibody molecules to very specific locations on sensitizable cells. This attachment process may involve the dissociation of antibody molecules from loose bonds with nonreactive cells and transfer to reactive cells. Experimental results have been presented which show that sensitization is related to the amount of precipitating antibody present in a mixture of antibody and homologous gamma-globulin. In the case of guinea pig atrial tissue, antibody probably is attached to several sites on reactive, histamine-releasing, cells.
Article
This chapter reviews an attempt to critically assess the important data available on the biological and physicochemical properties of reaginic antibodies for providing a clearer indication of their nature in relation to the rapidly increasing knowledge of immune antibodies. Application of the term “reagin” is restricted to the spontaneously produced skin-sensitizing antibodies found in allergic human sera. The corresponding antigen is referred to as “allergen,” no attempt being made to distinguish between the antigen responsible for the initial sensitization process and that evoking skin reactions in actively and passively sensitized humans. The more general term “allergic” has been preferred to “atopic” in referring to individuals showing immediate hypersensitivity. Possible modes of action of reagin such as antibodies with “dual specificity,” comparison with mode of action of anaphylactic-type antibodies, hypothetical reagin-allergen interaction model, a possible mechanism of reagin formation, nature of allergenic determinants, role of epithelial glycoprotein, and influence of nature of antigenic determinant on type of hypersensitive response evoked are also discussed in this chapter. Physicochemical characterization of reagins such as salt and ethanol fractionation studies, electrophoretic characteristics, and sedimentation characteristics is also provided in this chapter. Tissue studies and artificial induction of reagin-like antibodies is also discussed in this chapter.
Article
The morphological changes of the capillary endothelium in lung have been studied with administration of soluble immune complexes. With the administration of soluble immune complexes, there were noted vacuolarisation, lift-up phenomenon, arcade formation and subendo-thelial edema of pulmonary capillaries.
Article
SummaryA child with the syndrome of diabetes mellitus, insulin-resistance, Coombs' positive autoimmune hemolytic anemia, lymphadenopathy, and hepatosplenomegaly was described. The patient's IgG anti-insulin antibody was shown to be negative for passive cutaneous anaphylaxis (PCA). In as much as the patient was refractory to prednisone therapy and presented a negative PCA test, he was consistent with the Oakley hypothesis that insulin-resistant prednisone-insensitive patients will be negative for PCA. The half-life of the patient's red blood cells was shown to vary as a function of the type of insulin used in therapy. The child was shown to have immune-complexes in his serum of insulin-anti-insulin specificity, and it was suggested that these complexes are partially responsible for the microangiopathy that is so often observed in diabetes mellitus.
Article
In earlier studies we found that normal rat macrophages, preincubated at 37 degrees C with the serum of syngeneic rats immune to Schistosoma mansoni, strongly adhered to S. mansoni schistosomules whereas no significant adherence was induced with serum from normal rat. Using 51 chromium release assay, it now proved that immune serum-incubated macrophages lysed the schistosomules in 18 h. Absorption experiments and ultracentrifugation of the immune serum showed that immune complexes containing specific IgE antibody against S. mansoni and soluble parasite antigens induced macrophage adherence and cytotoxicity. Using various labeling techniques, the binding of aggregated rat IgE to the macrophage surface at 37 degrees C is evident in conditions where endocytosis is neglible. The binding of immune complexes containing IgE antibody to S. mansoni elicits dramatic ultrastructural changes in the macrophage and an increase of its lysosomal enzymes together with specific lytic activity for the schistosomules. The parallel development of immune serum-induced macrophage adherence or cytotoxicity with the level of circulating IgE antibody to the parasite in correlation with the development of immunity in the rat, suggests that this new mechanism of macrophage activation could play a role in immune effector mechanisms against S. mansoni. Therefore, IgE acts as a humoral mediator of cellular immunity.
Article
Experiments are described indicating that the magnitude and sensitivity of the passive cutaneous anaphylaxis (PCA) response in normal rats to a given level of immune reagents, may be enhanced by the addition of hemolytically active sera. A similar enhancement in normal rats has been obtained with C' component reagents possessing properties associated with the third component of C'. Parallelisms between in vitro fixation of C' and PCA induction by antigen and antibody are shown. The horse anti-pneumococcus system has low C'-fixing potencies and is also less efficient than the rabbit polysaccharide system in the induction of PCA. Findings of a similar nature were observed in the reaction of rabbit anti-ribonuclease with ribonuclease, the acetylated and guanidinated derivatives of the enzyme. The injection of hemolytically active serum into C'-deficient rats was accompanied by a partial restoration of PCA. Restorative effects were also noted with heated and ammonia-treated serum. The return of hemolytic potency and responsiveness to PCA in C'-depleted rats, follow a similar time course. The data presented indicate that the PCA reaction can be studied as a function of at least three variables, antigen, antibody, and a serum constituent resembling C'.
Article
Solutions of soluble complexes formed between bovine serum albumin (as antigen, Ag) and its rabbit antibodies (Ab) have been subjected to electrophoresis and ultracentrifugation over a range of pH. While the distribution of species in these solutions is apparently not grossly altered between pH 7.5 and 4.5, between pH 4.5 and 3.0 very extensive dissociation of the complexes occurs. Equilibrium constants, K, for the reaction Ag + Ab AgAb can be calculated as a function of pH. The variation of K with pH provides strong evidence that a single carboxyl group is involved in every Ag-Ab bond in this system.
Article
Solutions of soluble complexes formed between bovine serum albumin (as antigen) and its rabbit antibodies have been subjected to electrophoresis and ultracentrifugation. From these experiments it has been possible to obtain the equilibrium concentration of uncombined antigen (Ag) in a given solution of known total antigen and total antibody (Ab) content. With the aid of the Goldberg theory, this information has been interpreted to give equilibrium constants for the reaction: Ag + AgAb ⇄ (Ag)2Ab; K = (2.6 ± 0.5) × 104, ΔF0 = -5.5 ± 0.2 kcal., ΔH0 = 0 ± 2 kcal., and ΔS0 = +20 ± 8 e.u., for this reaction in this system in veronal buffer, pH 8.5, at 0°. The significance of these thermodynamic parameters is discussed.
Article
A quantitative ultracentrifugal and electrophoretic study has been performed with the soluble antigen-antibody complexes formed in antigen excess between lightly-iodinated bovine serum albumin antigen and precipitating rabbit anti-bovine serum albumin antibodies. A particular species of complex, the a complex, is found to predominate in large antigen excess, and it is demonstrated that this complex must be largely the species containing two antigen molecules and one antibody. It is concluded, therefore, that the antibodies in this system are largely bivalent.
Article
From a previous ultracentrifugal and electrophoretic study of the soluble complexes formed between lightly iodinated bovine serum albumin (antigen) and precipitating rabbit antibodies to bovine serum albumin, data are available concerning the amounts of free antigen present in equilibrium in solutions of known total antigen and antibody concentrations. These data are now used in conjunction with part of the theory recently developed by Goldberg for the antigen-antibody reaction, to test the theory, and to obtain an estimate of the free energy change in this reaction for the particular system studied.
Article
1. Normal rabbit serum and low titered antisera contain appreciable quantities of complement nitrogen even after 3 to 5 months of storage at 0 C. 2. Significant differences in both the total nitrogen precipitated from antisera with low concentrations of antibody and the antibody N/antigen N ratios before and after decomplementation of the antisera have been demonstrated. 3. The differences in immunochemical behavior of antisera diluted in normal rabbit serum and in 0.85% saline are markedly reduced after decomplementation of the diluting serum.
Article
The ultrasonic dosages given in W. J. Fry et al., "Ultrasonic lesions in the mammalian central nervous system" [Science 122, 517 (1955)] should be corrected as follows: For the lesion illusrated in Fig. 1, the dosage was 40 atm acoustic pressure amplitude and 3.9(10(2)) cm/sec acoustic particle velocity amplitude. For the lesions illustrated in Figs. 2 and 3, the dosage was 41 atm acoustic pressure amplitude and 4.0(10(2)) cm/sec acoustic particle velocity amplitude.
Article
Experiments are described indicating that the magnitude and sensitivity of the passive cutaneous anaphylaxis (PCA) response in normal rats to a given level of immune reagents, may be enhanced by the addition of hemolytically active sera. A similar enhancement in normal rats has been obtained with C' component reagents possessing properties associated with the third component of C'. Parallelisms between in vitro fixation of C' and PCA induction by antigen and antibody are shown. The horse anti-pneumococcus system has low C'-fixing potencies and is also less efficient than the rabbit polysaccharide system in the induction of PCA. Findings of a similar nature were observed in the reaction of rabbit anti-ribonuclease with ribonuclease, the acetylated and guanidinated derivatives of the enzyme. The injection of hemolytically active serum into C'-deficient rats was accompanied by a partial restoration of PCA. Restorative effects were also noted with heated and ammonia-treated serum. The return of hemolytic potency and responsiveness to PCA in C'-depleted rats, follow a similar time course. The data presented indicate that the PCA reaction can be studied as a function of at least three variables, antigen, antibody, and a serum constituent resembling C'.
Article
The retention of antigen in rabbit liver tissue, resulting from a primary intravenous injection, is influenced by immunization brought about by subsequent intravenous injections of the same antigen. In rabbits given a single primary intravenous injection of radioactive antigen, the retention of radioactivity in liver tissue, after a period of 21 days, was greater than when the primary injection was followed by secondary injections of the same, but non-radioactive antigen. The results were similar for both S^(35)-azohemocyanin and S^(35)-azo-bovine-serum-albumin, except the hemocyanin was retained to a greater extent than the albumin. There was very little if any correlation between the number of secondary injections and retention of the initial injection. Quantitative antibody nitrogen data, obtained for the serum of each rabbit showed, in general, an inverse relationship between circulating antibody and radioactivity retained, i.e. the higher the circulating antibody titer, the lower the retention of radioactivity in liver tissue. Passively administered homologous antibody did not produce a change in the retention of the primary injection of antigen nor did secondary injections of a heterologous native protein injected according to the same immunization schedule as the homologous azoprotein. From these results it may be concluded that an intracellular antibody-forming activity influences the loss (or retention) of antigen deposited in liver tissue and that the mechanism is immunologically specific.
Article
Immediate skin reactions were obtained in normal guinea pigs by intradermal injections of soluble antigen-antibody complexes. Immediate skin reactions were also obtained by injections of an electrophoretically separated component of serum from immunized or normal rabbits.
Article
1) Soluble BSA anti-BSA complexes formed in regions of extreme antigen excess are less anaphylactigenic for the mouse than similar complexes formed in regions of low antigen excess. 2) The hypothesis is advanced that anaphylactigenic activity of these complexes for the mouse is dependent upon their capacity to react with complement.