Larralde / Willms / Ortiz-Ortiz Molecules, Cells, and Parasites in Immunology

1. Auflage 2014
ISBN: 978-1-4832-7026-5
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: 6 - ePub Watermark

E-Book, Englisch, 246 Seiten

Molecules, Cells, and Parasites in Immunology contains the proceedings of a symposium on immunology held in Mexico City in the fall of 1979 under the auspices of the National University of Mexico (UNAM). This volume brings together prominent foreign scientists and Mexican investigators to discuss the areas of immunology that being developed at UNAM. Either in the form of reviews or of original presentations, the various authors proffer observations and ideas that generally relate to the regulation of the immune response at the molecular and cellular levels. The topic of Molecules encompasses the effect of molecular aggregation upon the triggering of physiological events, cooperative binding, and the expression of immunoglobulins during the lymphocyte differentiation process. Under the heading of Cells, there are contributions dealing with the physiological and pathological roles of interiorized antibodies; the mechanisms of antigen recognition by T cells; and the synthesis of membrane proteins by myeloma cells which somehow protect them from killer T cells. The section on Parasites includes some unique contributions on the immunology of human and animal cysticercosis and amebiasis, complemented by a general discussion of host-parasite relationships.

Autoren/Hrsg.

Larralde, Carlos

Willms, Kaethe

Ortiz-Ortiz, Librado

Inhaltsverzeichnis

1;Front Cover;1
2;Molecules, Cells, and Parasites in Immunology;4
3;Copyright Page;5
4;Table of Contents;6
5;CONTRIBUTORS;8
6;PREFACE;12
7;ACKNOWLEDGMENTS;14
8;CHAPTER 1. MOLECULAR MECHANISMS OF ANTIBODY ACTION: USE OF CROSS-LINKING REAGENTS;16
8.1;I. INTRODUCTION;16
8.2;II. NEW STUDIES;17
8.3;III. CONCLUDING STATEMENT;24
8.4;REFERENCES;25
9;CHAPTER 2.
MEMBRANE-RECEPTOR MECHANISMS IN ANAPHYLAXIS;28
9.1;I. THE CONCEPT OF A RECEPTOR;28
9.2;II. EXCITABLE TISSUE ANAPHYLAXIS;30
9.3;III. CONNECTIVE TISSUE INVOLVEMENT;36
9.4;IV. TWO DISTINCT RECEPTOR COMPONENTS;40
9.5;ACKNOWLEDGMENTS;41
9.6;REFERENCES;42
10;CHAPTER 3.
COOPERATIVITY AND ANTIBODY FUNCTION;44
10.1;I. INTRODUCTION;44
10.2;II. EXPERIMENTAL RESULTS;45
10.3;III. COOPERATIVITY OR ARTIFACT?;47
10.4;IV. ESTIMATION OF THE PARAMETERS OF COOPERATIVITY;52
10.5;V. PREVIOUS OBSERVATIONS;55
10.6;VI. DISCUSSION;55
10.7;VII. CONCLUDING REMARKS;56
10.8;ACKNOWLEDGMENTS;56
10.9;REFERENCES;56
11;CHAPTER 4.
DIFFERENTIATION OF THE B-LYMPHOCYTE;58
11.1;I. INTRODUCTION;58
11.2;II. GENERATION OF B-LYMPHOCYTES;59
11.3;III. THE PRE-B CELL;59
11.4;IV. THE IMMATURE B-CELL;60
11.5;V. DEVELOPMENT OF HEAVY CHAIN CLASS (ISOTYPE) HETEROGENEITY;62
11.6;VI. THE PROBLEM OF MULTIPLE CLASS REPRESENTATION;63
11.7;VII. THE ROLE OF IgD;64
11.8;VIII. MODE OF MEMBRANE INSERTION OF SURFACE IMMUNOGLOBULIN;64
11.9;ACKNOWLEDGMENT;65
11.10;REFERENCES;65
12;CHAPTER 5. ANTIBODY PENETRATION INTO LIVING CELLS:
MECHANISMS AND CONSEQUENCES;68
12.1;I. INTRODUCTION;68
12.2;II. ANTIBODY PENETRATION AND THE Fc RECEPTOR;69
12.3;III. EVIDENCE OF ANTIBODY PENETRATION IN VIVO;71
12.4;IV. CONSEQUENCES OF ANTIBODY PENETRATION;73
12.5;V. ROLE OF ANTIBODY PENETRATION IN IMMUNOBIOLOGY;76
12.6;ACKNOWLEDGMENT;77
12.7;REFERENCES;77
13;CHAPTER 6. LAMBDA LIGHT CHAINS
AND IDIOTYPE-BEARING T CELLS;80
13.1;I. INTRODUCTION;80
13.2;II. . LIGHT CHAINS;82
13.3;III. Id+ ANTI-NP T CELLS;84
13.4;IV. DEPENDENCE OF THE SPECIFICITY OF ANTI-Id ANTIBODIES ON VL DOMAINS OF THE Id-BEARING MOLECULE;86
13.5;V. LOGISTICS;87
13.6;REFERENCES;88
14;CHAPTER 7. CELLULAR RECOGNITION IN TUMOR IMMUNOLOGY: TUMOR RESISTANCE TO IMMUNE DESTRUCTION
BY CYTOTOXIC T LYMPHOCYTES;90
14.1;I. INTRODUCTION;90
14.2;II. MATERIALS AND METHODS;91
14.3;III. RESULTS;93
14.4;IV. DISCUSSION;101
14.5;REFERENCES;103
15;CHAPTER 8.
CHEMOTAXIS;106
15.1;I. INTRODUCTION;106
15.2;II. CHEMOTAXIS: GENERAL AND TECHNICAL CONSIDERATIONS;107
15.3;III. CHEMOTAXIS IN HUMAN NEWBORNS AND AMEBIASIS;111
15.4;REFERENCES;115
16;CHAPTER 9. BIOCHEMICAL CHARACTERIZATION OF LYMPHOCYTE MEDIATOR-MACROPHAGE INTERACTIONS;118
16.1;I. INTRODUCTION;118
16.2;II. RESULTS AND DISCUSSION;120
16.3;REFERENCES;126
17;CHAPTER 10. B LYMPHOCYTE STIMULATION BY PARASITIC ORGANISMS;128
17.1;I. INTRODUCTION;128
17.2;II. METHODS;129
17.3;III. RESULTS;129
17.4;IV. DISCUSSION;134
17.5;ACKNOWLEDGMENT;137
17.6;REFERENCES;137
18;CHAPTER 11. THE IMMUNOLOGY OF HUMAN CYSTICERCOSIS;140
18.1;I. INTRODUCTION;140
18.2;II. IMMUNODIAGNOSIS OF BRAIN CYSTICERCOSIS;141
18.3;III. SEROEPIDEMIOLOGY;144
18.4;IV. HUMORAL IMMUNE RESPONSE;148
18.5;V. IMMUNITY TO CYSTICERCOSIS;152
18.6;VI. CELLULAR IMMUNE RESPONSE;154
18.7;VII. HOST-PARASITE INTERFACE;154
18.8;VIII. CONCLUSIONS;154
18.9;REFERENCES;156
19;CHAPTER 12.
IMMUNOPATHOLOGY OF CYSTICERCOSIS;160
19.1;I. HOST-PARASITE SPECIFICITY;160
19.2;II. ULTRASTRUCTURE OF THE PARASITE SURFACE;162
19.3;IIII. SYNTHESIS OF PARASITE PROTEINS;166
19.4;IV. HOST RESPONSE;168
19.5;ACKNOWLEDGMENTS;176
19.6;REFERENCES;176
20;CHAPTER 13.
IMMUNOLOGY OF AMEBIASIS;178
20.1;I. INTRODUCTION;178
20.2;II. HUMORAL IMMUNE REACTIONS;178
20.3;III. CELLULAR IMMUNE REACTIONS;181
20.4;IV. IMMUNITY AND ANTIAMEBIC IMMUNIZATION;184
20.5;REFERENCES;190
21;CHAPTER 14.
THE HOST-PARASITE INTERACTION;194
21.1;I. INTRODUCTION;194
21.2;II. SUSCEPTIBILITY TO INFECTION;194
21.3;III. IMMUNOSUPPRESSION AND IMMUNOPOTENTIATION;197
21.4;IV. ANTIGENS;199
21.5;V. CONCLUSIONS;201
21.6;REFERENCES;202
22;CHAPTER 15.
THE ROLE OF CIRCUMSTANCE IN IMMUNITY;206
22.1;I. INTRODUCTION;206
22.2;II. CIRCUMSTANCE AND IMMUNITY AT THE LEVEL OF A COMMUNITY;207
22.3;III. CIRCUMSTANCE AND IMMUNITY AT THE LEVEL OF AN ORGANISM;213
22.4;IV. CIRCUMSTANCE AT THE CELLULAR LEVEL;218
22.5;V. CIRCUMSTANCE AT THE MOLECULAR LEVEL;220
22.6;VI. DISCUSSION;225
22.7;REFERENCES;228
23;CHAPTREC 16. HEMICAL SYNTHESIS FOR THE UNDERSTANDING OF IMMUNE RESPONSE PHENOMENA AND FOR THEIR MEDICAL APPLICATION;230
23.1;I. INTRODUCTION;230
23.2;II. SYNTHETIC ANTIGENS WITH PEPTIDE DETERMINANTS OF DEFINED SEQUENCE;231
23.3;III. SYNTHETIC PEPTIDE OF CARCINOEMBRYONIC ANTIGEN OF THE COLON;236
23.4;IV. SUPPRESSION OF EAE AND POSSIBLY OF MULTIPLE SCLEROSIS WITH A BASIC SYNTHETIC COPOLYMER;238
23.5;V. ANTIVIRAL EFFECT ON MS-2 COLIPHAGE OBTAINED WITH A SYNTHETIC ANTIGEN;239
23.6;VI. CHALLENGE OF THE COMBINED USE OF SYNTHETIC ANTIGENS AND SYNTHETIC ADJUVANTS;241
23.7;VII. CONCLUDING REMARKS;241
23.8;REFERENCES;242
24;INDEX;244

Leseproben

MOLECULAR MECHANISMS OF ANTIBODY ACTION: USE OF CROSS-LINKING REAGENTS*

HENRY METZGER, HELEN HARTMANN, DAVID HOLOWKA and CLARE FEWTRELL,     Section on Chemical Immunology, Arthritis and Rheumatism Branch, National Institute of Arthritis Metabolism and Digestive Diseases, National Institutes of Health, Bethesda, Maryland, U.S.A.

Publisher Summary

This chapter discusses the molecular mechanisms of antibody action and describes the use of cross-linking reagents. Antibodies have no ability to alter the antigen with which they combine directly; they change the ultimate fate of the antigen only by interacting with effector system. When the high molecular weight product of cross-linking with the cleavable reagent is rerun under conditions where the crosslinks are broken, a new component is observed that has a molecular weight of about 30,000. The cross-linked product can be observed after reacting either intact cells or cells solubilized with nonionic detergents. Some of the serotonin that can be released by the higher oligomers is completely resistant to release by dimers. Dimers by themselves are adequate to stimulate secretion of at least a portion of the releasable serotonin. Trimers are more active than dimers, while the higher oligomers are even more effective.

I INTRODUCTION

Immunologists have traditionally delighted in clumping things together. This is reflected in the by-now classical methods of precipitin analysis and agglutination as well as in the new clumping methods which are constantly being developed. Lately, ‘rosette’ formation has become a popular technique among immunologists, and most recently, the use of ‘patching’ and ‘capping’ and of cell fusion to make ‘hybridomas’ demonstrates that the tradition lives on.

It is interesting to speculate that the immunologist’s early use of aggregation as a favored analytical tool stemmed from a subconscious prescience that aggregation was of fundamental importance to what the immunologist was interested in studying; namely, the mechanisms of immune responsiveness. Whether one accepts this psycho-immunologic thesis or not, the significance of aggregation in immunologic mechanisms cannot be disputed. At the cellular level, direct cell to cell interactions have been implicated in a variety of phenomena; e.g., T cell interaction with macrophages and B cells, and T cell killing. Similarly, the significance of aggregation in antibody-mediated mechanisms is well recognized. After some brief comments on the latter subject, we shall describe how still another technique of clumping – the use of chemical cross-linking reagents – has permitted us to explore some basic questions in the system with which we are working.

Antibodies have no ability to alter the antigen with which they combine directly; they change the ultimate fate of the antigen only by interacting with effector systems. In some instances, this leads to a direct and prompt change in the antigen’s prospects, as in phagocytosis. Alternatively, a much more indirect pathway is chosen, such as stimulation of antibody-forming precursor cells. The mechanisms by which antibodies mediate these effects have received considerable attention, especially in recent years, as our detailed knowledge of antibody structure has increased. Several plausible models have been proposed and discussed in reviews [1–3]. A recent workshop, the first to discuss this topic comprehensively [4], did not come up with a consensus report. Nevertheless, with only one significant exception – the observations of Koshland and her colleagues [5,6] – all of the data on activation of effector systems by antibodies point to the critical requirement for aggregation of antibody; specifically, of the Fc regions of the antibodies. Whether the findings of Koshland’s group (that apparently univalent antigens induce complement fixation by IgM) are a valid exception is uncertain. Univalent ligands assuredly don’t cause intermolecular aggregation, but it is less clear whether the special antigens required to mediate the effects studied by these investigators cause intramolecular aggregation of the Fc pentamers in IgM.

In no system is the need for aggregation of antibody more persuasively supported than in the IgE-mast cell system [7–9]. In this instance, antigen reacts with cell-bound IgE (alternatively, preformed IgE-antigen complexes become cell-bound), resulting in the triggering of noncytotoxic degranulation. Any procedure which serves to aggregate the surface IgE will cause stimulation, although hyper-aggregation is inhibitory. The reason why aggregation of the IgE is critical has recently been elucidated by the use of antibodies directed to the cellular membrane component (receptor for IgE) to which the IgE is bound [10,11]. These studies showed that what is important is that the becomes aggregated. Even cells grown in the total absence of IgE can be stimulated to secrete with bivalent antireceptor antibodies [11].

II NEW STUDIES

We shall now describe two studies recently completed in our laboratory which have used chemical cross-linking procedures as a probe for studying how receptor aggregation may mediate mast cell (or basophil) secretion [12,13]. The reagents used in these studies (Fig. 1) are somewhat more useful than the one traditionally used by immunologists – bisdiazobenzidine. The latter compound is sufficiently carcinogenic to make its use banned in the United States; in addition, the newer reagents are more stable, specific, and versatile [14]. The versatility arises because one can vary the length between the reactive groups as well as prepare analogs which can be cleaved under mild conditions. The latter property has been particularly useful for some of our work.

FIG. 1 Chemical cross-linking reagents used in the experiments discussed in this paper. The chemistry of these reagents is reviewed in Ref. 14.

A Effect of IgE Oligomer Size in the Triggering of Mast Cells

The first study made use of cross-linking reagents to analyze the extent of receptor aggregation that is required for optimal stimulation of mast cell degranulation. Our original work in this area involved the use of dimethylsuberimidate (DMS) to prepare well-defined stable oligomers of rat IgE [15]. These were then used to elicit passive cutaneous anaphylactic reactions in the rat. These experiments led us to conclude that dimers of IgE could generate so-called ‘unit signals’. It might take many such signals to trigger a cell completely, but the individual signal could be generated by as small a unit as a receptor dimer.

What was not entirely clear from these studies was whether higher oligomers might yield stronger signals. While the data suggested that they did not, the data were less definitive. Furthermore, a recent investigation using oligomers of human IgE on human peripheral blood basophils studied has suggested that all oligomers may not be equal [A. Sobotka and L. Lichtenstein, submitted for publication].

In an attempt to resolve this question in our system, rat IgE was reacted with DMS and separated into monomers, dimers, trimers, and higher oligomers by gel filtration (Fig. 2A). We principally studied rat basophilic leukemia (RBL) cells [16], which can be grown as solid tumors in animals or in cell culture and are a unique resource for detailed structural analyses such as those described below. The cells were allowed to incorporate radioactive serotonin and the stimulated release of the latter by the oligomers was then assayed (Fig. 2B). Our experiments will be published in complete form elsewhere [12], and only the most significant findings will be summarized here:

FIG. 2 Separation of IgE oligomers and assay of their relative activities. A A trace amount of iodinated IgE was added to nonradioactive IgE (21 mg, 60 mg/ml) and the solution reacted with a 16-fold molar excess of DMS for 1 h at 30°C in 0.2 M Tris buffer, pH 8.6. The preparation was applied to sequential Sephadex G-200 and Ultrogel Ac A 22 columns, and the radioactivity in the effluent was measured. The peaks are labeled to indicate the relative effluent volumes for monomer (M), dimer (D), trimer (T), and higher oligomers (H). B Analysis of column fractions (numbers on the right) for their capacity to stimulate the release of incorporated tritiated serotonin (3H.5HT) from RBL cells. The test material was incubated with 2 × 106 cells/ml for 1 h at 37°C. The medium contained 1.8 mM Ca++ but not D2O.

(1) Dimers by themselves were adequate to stimulate secretion of at least a portion of the releasable serotonin, although this was often barely observable in the absence of D2O (Fig. 2B).

(2) Trimers were more active than dimers, while the higher oligomers were even more effective (Fig. 2B, Table I).

TABLE I

Serotonin Release by Oligomers of...