Advances in Immunology Volume 13

ADVANCES I N

Immunology

VOLUME 13

CONTRIBUTORS TO THIS VOLUME ELMERL. BECKER

HANSBENNICH BARRYR. BLOOM A. D. M. BRYCESON JOHN

E. HOPPER

S. GUNNAR0. JOHANSSON

ALFREDNISONOFF

J. L. TURK

ADVANCES IN

Immunology EDITED BY

F. J. DIXON, JR.

H E N R Y G. KUNKEL

Division of Experimenfal Pathology Scripps Clinic and Research Foundafion

The Rockefeller Universify N e w York, N e w York

La Jolla, California

V O L U M E 13 1971

ACADEMIC PRESS

New York and London

COPYRIGHT 8 1971, BY ACADEMIC PRESS, INC. ALL RIGHTS RESERVED NO PART OF THIS BOOK MAY BE REPRODUCED IN A N Y FORM, BY PHOTOSTAT, MICROFILM, RETRIEVAL SYSTEM, OR ANY OTHER MEANS, WITHOUT WRITTEN PERMISSION PROM THE PUBLISHERS.

ACADEMIC PRESS, INC. 111 Fifth Avenue, New

York, New York 10003

United Kingdom Edition published b y ACADEMIC PRESS INC. (LONDON) LTD.

24/28 Oval Road, London)NWI IDD

LIBRARY OF CONGRESS CATALOG CARD

NUMBER: 61-17057

PRINTED IN THE UNITED STATES OF AMERICA

CONTENTS LIST OF COSTRIBUTORS. PREFACE

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

vii ix

CONTENTS OF PREVIOUS VOLUhlES

xi

Structure and Function of Human Immunoglobulin E

HANSBESNICHA N D S. GUSNAR0. JOHANSSON

I. Introduction . . . . . . . . . . . . 11. Isolation and Physicochemical Characteristics of Inimunoglobulin E 111. Properties of Antigenically and Biologically Active Structural Regions of Immunoglobulin E . . . . . . . . IV. Methods for Determination . . . . . . . . . V. Levels of Immunoglobulin E in Healthy Individuals . VI. Levels of Inimunoglobulin E in Disease . . . . . . VII. Detection of Antibody Activity in the Immunoglobulin E Class . VIII. Metabolism . . . . . . . . . . . . IX. Concluding Remarks . . . . . . . . . . References . . . . . . . . . . .

1 2

19 28 29 35 45 49 51 51

. .

.

Individual Antigenic Specificity of Immunoglobulins

JOHNE. HOPPERAND ALFRED NISONOFF

I. Introduction . . . . . . . . . . . . 11. Individual Antigenic Specificities in Monoclonal Proteins . . . 111. Individual Antigenic Specificities in Antibody Populations . . IV. Cross-Reactions of Antiidiotypic Sera and Evidence for Identical . . . . . Molecules in Different Individual Animals V. Evidence Based on Idiotypic Specificity for Limited Heterogeneity of Normal Antibody Populations . . . . VI. Persistence and Changes of Antibody Populations during Prolonged Immunization . . . . . . . . . VII. Shared Idiotypic Determinants in IgC and IgM Antibodies of the Same Specificity . . . . . . . . . . VIII. Localization of Individually Specific Antigenic Determinants . IX. Cross-Reactions of Anti-ind Antibodies with Nonspecific Immunoglobulins . . . . . . . . . . . X. Monoclonal Origin of Molecules with Individually Specific Antigenic Determinants . . . . . . . . . XI. Summary . . . . . . . . . . . . References . . . . . . . . . . . .

58 60 63 69

.

75 76 81 85 92 94 95 97

In Vitro Approaches to the Mechanism of Cell-Mediated Immune Reactions

BARRYR. BLOOM

I. Introduction . . . . 11. Lymphocyte Transformation

. .

.

. V

.

.

.

.

.

.

. .

. .

.

.

. .

102 104

vi

CONTENTS

.

111 Direct Cytotoxicity of Target Cells by Lymphocytes . . . IV . Mediators-Qualitive Basis of the Response . . . . . V. Enumeration of Specifically Sensitized Cells-Quantitative Basis of the Response . . . . . . . . . . VI . Reality Testing-Relationships between in Vitro Results and CellMediated Immunity in Vioo . . . . . . . . VII . Relationships between Cell-Mediated Immunity and Antibody Formation . . . . . . . . . . References . . . . . . . . . . . .

.

.

111 122

.

160

.

169

.

178 193

.

Immunological Phenomena in leprosy and Related Diseases

.

.

J . L TURKAND A . D M . BRYCESON

. . . . . . . . . . . I . Introduction . I1. Leprosy . . . . . . . . . . . . . 111. Leishmaniasis . . . . . . . . . . . . IV . Concept of a Host-Determined Spectrum of Clinical Manifestations in Other Chronic Infections in Man . . . . . . . References . . . . . . . . . . . .

.

. .

209 210 237

.

259 261

.

267 270 271 285 286 289 291 298 299 305 307 308

.

Nature and Classification of Immediate-Type Allergic Reactions

.

ELMERL BECKER

. .

I Introduction . . . . . . . . . I1 Sensitization . . . . . . . . . 111. Components of the Allergic Reaction . . . . IV . Sites of the Antigen-Antibody Reaction . . . V. Time Course of Allergic Reactions . . . . VI The Terrain . . . . . . . . . VII . Basis and General Description of the Classification VIII Direct Responses (Non-mediator Determined) . . IX . Indirect Responses (Mediator Determined) . . X . Mixing of Categories in Natural Reactions . . XI Pseudoallergic Reactions . . . . . . References . . . . . . . . .

. .

.

AUTHOR INDEX .

.

.

.

.

.

.

.

.

.

SUBJECTINDEX .

.

.

.

.

.

.

.

.

.

.

.

.

. . . . .

. . . . .

. . . . .

. . . . .

. .

. .

. .

. .

. .

. . . .

. .

. . . .

. .

. . . .

. .

. . . .

315 333

LIST OF CONTRIBUTORS Numbers in parentheses indicate the pages on which the authors’ contributions begin.

ELMERL. BECKER, Department of Pathology, University of Connecticut Schools of Medicine and Dentistry, Farmington, Connecticut (267) HANS BENNICH, The Wallenberg Laboratory, University of Uppsala, Uppsala, Sweden (1) BARRYR. BLOOM, Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York (101) A. D. M. BRYCESON, Department of Medicine, Ahmadu Bello University, Zaria, Nigeria (209)

E. HOPPER, Department of Medicine, Pritzker School of Medicine, University of Chicago, Chicago, Zllinois ( 5 7 )

JOHN

S. GUNNAR0. JOHANSSON, The University Blood Center, University of Uppsala, Uppsala, Sweden (1) ALFREDNISONOFF, Department of Biological Chemistry, University of Zllinois College of Medicine, Chicago, Zllinois (57)

J. L. TURK,Department of Pathology, Royal College of Surgeons of England, Lincoln’s Inn Fields, London, England ( 2 0 s )

vii

This Page Intentionally Left Blank

PREFACE It can safely be predicted that the seventies will be the decade of cellular immunology. Already the expansion is cvideiit on all sides and many immunologists, previously involved in the antibody field, are turning to cellular work. To somc cxtent this may be unfortunate because that most basic of problcms, thc mechanism of antibody variability, remains an enigma. Immunologists are still evenly divided on the issue of whether the germ line theory or a typc of somatic model holds the explanation. Possibly the work at thc cellular level might provide the answer. The Advances will participate actively in this timely movement and, as exemplificd by this volumc, will also continue to be involved in the currently lcss popular but no lcss important areas of immunology, The articlc by Drs. John E. Hopper and Alfred Nisonoff concerns that very uscful label of the immunoglobulins, their individual antigenic specificity or idiotypic specificity. The authors have utilized this property in superb fashion to tracc the devclopment of different antibody producing clones of cells in the primary and secondary response. It is abundantly clear that antigens related to thc V regions aiid antibody combining sites are followed in thesc studies. There are few areas where problcms of iiomcnclature are more varied and confusing than in thc field of allergic reactions. Dr. Elmer L. Becker treats this subject from all aspects, ranging from the initiating antigen, through the mediators produced, to the final cell involvement. A very reasonable classification of immediate-type allergic reactions has emerged that takes into account the many different phases of these reactions. One of the exciting chaptcrs in immunology has been the recent dclineatioii of the IgE class of iniiiiuiioglobulins and the demonstration of its significance for atopic allergic disorders. Just as in all other areas of immunoglobulin work, the discovery of a myeloma protein of the IgE typc contributed cnorinously to thc successful evolution of this work. Drs. Hans Bennich and S. G~iiiiiar0. Johansson wcre responsible for this important aspcct and they not only review this field but also present many observations that have not been published elsewhere. Because of thc low concentration of IgE in most scra, its measurement has prescnted a special challenge. Thc ingcnious procedures devcloped by the authors, as me11 as other methods, arc discussed in useful detail. Drs. J. L. Turk and A. D. hl. Bryceson review the various different immunological reactions to the specific organisms in leprosy and ix

X

PREFACE

leishnianiasis. These authors have played a primary role in interpreting these reactions in terms of modern concepts of immunology. Defects in cellular immunity clearly play a major role in special forms of these disorders and many of the principles derived from these studies hold implications for a number of other diseases. Dr. Barry R. Bloom, one of the leaders in the cellular immunity expansion, describes some of the forefronts of this field. The inany mediators involved in lymphocyte reactions are considered in special detail. None of these factors has been isolated in pure form, which will be essential for their eventual understanding. However, an overall picture of the intricacies of cellular iminunity is beginning to emerge which relates the various experimental models to in vivo events. The cooperation and valuable assistance of the publishers in the production of Volume 13 are gratefully acknowledged. H. G . KUNKEL F. J. DIXON July 1971

Contents of Previous Volumes Volume 1 Transplantation Immunity and Tolerance

M. HA~EK, A. LENGEROVA, AND T. HRABA Immunological Tolerance of Nonliving Antigens

RICHARDT. SMITH Functions of the Complement System

ABRAHAMG. OSLER In Vitro Studies of the Antibody Response

ABRAMB. STAVITSKY Duration of Immunity in Virus Diseases

J. H. HALE Fate and Biological Action of Antigen-Antibody Complexes

WILLIAM 0. WEIGLE Delayed Hypersensitivity to Simple Protein Antigens

P. G. H. GELLAND B. BENACERF~AF The Antigenic Structure of Tumors

P. A. GORER AUTHORINDEX-SUB JECX INDEX Volume 2 Immunologic Specificity and Molecular Structure

FREDKARUSH Heterogeneity of y-Globulins JOHN

L. FAHEY

The Immunological Significance of the Thymus

J. F. A. P. MILLER,A. H. E. MARSHALL, AND R. G. WHITE Cellular Genetics of Immune Responses

G. J. V. NOSSAL Antibody Production by Transferred Cells

CHARLESG. COCHRANE AND FRANK J. DIXON Phagocytosis

DERRICK ROWLEY xi

xii

CONTENTS OF PREVIOUS VOLUMES

Antigen-Antibody Reactions in Helminth Infections

E. J. L. SOUISBY Embryological Development o f Antigens

REED A. FLICKINCER AUTHOR INDEX-SUB JECT INDEX Volume 3 In Vitro Studies of the Mechanism of Anaphylaxis

K. FRANK AUSTENAND JOHN H. HUMPHREY The Role of Humoral Antibody in the Homograft Reaction

CHANDLER A. STETSON Immune Adherence

D. S. NELSON Reaginic Antibodies

D. R. STANWORTH Nature of Retained Antigen and Its Role in Immune Mechanisms

DANH. CAMPBELL AND JUSTINE S. GARVEY Blood Groups in Animals Other Than Man

W. H. STONEAND M. R. IRWIN Heterophile Antigens and Their Significance in the Host-Parasite Relationship

C. R.

JENKIN

AUTHOR INDEX-SUB j ~ c INDEX r Volume 4 Ontogeny and Phylogeny o f Adoptive Immunity

ROBERTA. GOODAND BENW. PAPERMASTER Cellular Reactions in Infection

EMANUEL SUTERAND HANSRUEDY RAMSEIER Ultrastructure of Immunologic Processes JOSEPH

D. FELDMAN

Cell W a l l Antigens of Gram-Positive Bacteria

MACLYNMCCARTY AND STEPHEN I. MORSE Structure and Biological Activity of Immunoglobulins SYDNEY COHEN AND

RODNEYR. PORTER

CONTENTS OF PREVIOUS VOLUXZES

Autoantibodies and Disease

H. G. KUKKELAND E. M. TAN Effect of Bacteria and Bacterial Products on Antibody Response

J. MUNOZ AUTHOR INDEX-SUBJECT INDEX Volume 5 Natural Antibodies and the Immune Response

STEPHENV. BOYDEN Immunological Studies with Synthetic Polypeptides

MICHAELSELA Experimental Allergic Encephalomyelitis and Autoimmune Disease

PHILIPY. PATERSON The Immunology of Insulin

C. G. POPE Tissue-Specific Antigens

D. C. DUNONDE AUTHOR INDEX-SUB j ~ c INDEX r Volume 6 Experimental Glomerulonephritis: Immunological Events and Pathogenetic Mechanisms

EMILR. UNAMJEAND FRANK J. DIXON Chemical Suppression of Adaptive Immunity

ANNE. GABRIELSON AND ROBERT A. GOOD Nucleic Acids as Antigens

OTTOJ. PLESCIA AND WERNER BRAWN In Vifro Studies of Immunological Responses of lymphoid Cells

RICHARDW. DUTTON Developmental Aspects of Immunity JAROSL.4V

STERZLAND ARTHURM.

SILVERSTEIN

Anti-antibodies

PHILIPG. H. GELLAND ANDREWS. KELUS Conglutinin and lmmunoconglutinins

P. J. LACHMANN AUTHORINDEX-SUB JECT IXDEX

...

Xlll

xiv

CONTENTS OF PREVIOUS VOLUMES

Volume 7 Structure and Biological Properties of Immunoglobulins

SYDNEY COHENAND CESAR MILSTEIN Genetics of Immunoglobulins in the Mouse

MICHAELPOTTER AND ROSELIEBERMAN Mimetic Relationships between Group A Streptococci and Mammalian Tissues JOHN

B. ZABRISKIE

Lymphocytes and Transplantation Immunity

DARCY B. WILSON AND R. E. BILLINGHAM Human Tissue Transplantation

JOHNP. MERRILL AUTHOR INDEX-SUB J E INDEX ~ Volume 8 Chemistry and Reaction Mechanisms of Complement

HANSJ. MULLER-EBERHARD Regulatory Effect of Antibody on the Immune Response

JONATHAN W. UHRAND GORANMOLLER The Mechanism of Immunological Paralysis

D. W. DRESSER AND N. A. MITCHISON In Vifro Studies of Human Reaginic Allergy

ABRAHAMG. OSLER, LAWRENCE M. LICHTENSTEIN, AND DAVID A. LEVY AUTHOR INDEX-SUB JECT INDEX Volume 9 Secretory Immunoglobulins

THOMAS B. TOMASI, JR.,AND JOHNBIENENSTOCK Immunologic Tissue Injury Mediated by Neutrophilic leukocytes

CHARLES G. COCHRANE The Structure and Function of Monocytes and Macrophages

ZANVIL A. COHN The Immunology and Pathology of NZB Mice

J. B. HOWIEAND B. J. HELYER AUTHOR INDEX-SUB J E INDEX ~

CONTENTS OF PREVIOUS VOLUMES

Volume 10 Cell Selection by Antigen in the Immune Response

GREGORY W. SISKINDAND BARUJBENACERRAF Phylogeny of Immunoglobulins

HOWARD M. GREY Slow Reacting Substance of Anaphylaxis

ROBERTP. ORANGEAND K. FRANK AUSTEN Some Relationships among Hemostasis, Fibrinolytic Phenomena, Immunity, and the Inflammatory Response

OSCARD. RATNOFF Antigens o f Virus-Induced Tumors

KARL HMEL Genetic and Antigenetic Aspects of Human Histocompatibility Systems

D. BERNARDAMOS AUTHORINDEX-SUB JECT INDEX Volume 1 1 Electron Microscopy of the Immunoglobulins

N. MICHAELGREEN Genetic Control of Specific Immune Responses

HUGH0. MCDEVITTAND BARUJBENACERRAF The lesions in Cell Membranes Caused by Complement JOHN

H. HUMPHREY AND ROBERTR. DOURMASHKIN

Cytotoxic Effects of lymphoid Cells In Vitro

PETERPERLMANN AND GOFIANHOLM Transfer Factor

H. S. LAWRENCE Immunological Aspects of Malaria Infection

IVORN. BROWN AUTHORINDEX-SUB JECT INDEX Volume 12 The Search for Antibodies with Molecular Uniformity

RICHARDM. KRAUSE Structure and Function of y M Macroglobulins

HENRYMETZGER

xv

XVi

CONTENTS OF PREVIOUS VOLUMES

Transplantation Antigens

R. A. REISFELDAND B. D. KAHAN The Role of Bone Marrow in the Immune Response

NABIHI. ABDOUAND MAXWELLRICHTER Cell Interaction in Antibody Synthesis

D. W. TALMAGE, J. RADOVICH,AND H. HEMMINGSEN The Role of Lysosomes in Immune Responses

GERALDWEISSMANN AND PETERDUKOR Molecular Size and Conformation of Immunoglobulins

KEITH J. DORRINGTON AND CHARLES TANFORD AUTHORINDEX-SUB JECT INDEX

Structure and Function of Human Immunoglobulin E HANS BENNICH AND S . GUNNAR 0 . JOHANSSON The Wallenberg laboratory and The University Blood Center. University o f Uppsala. Uppsala. Sweden

.

I Introduction . . . . . . . . . . . I1. Isolation and Physicocheniical Characteristics of Immunoglobulin E . . . . . . . . . . A . Identification of Myeloma Protein N D . . . . . B Isolation . . . . . . . . . . . C . Chemical and Physical Characteristics . . . . . I11. Properties of Antigenically and Biologically Active Structural Regions of Immunoglobulin E . . . . . . . A . Properties of e Chains . . . . . . . . B . Properties of Enzymatic Fragments . . . . . IV . Methods for Determination . . . . . . . . V Levels of Immunoglobulin E in Healthy Individuals . . . A . Levels in Serum . . . . . . . . . B. Levels in Secretions . . . . . . . . . VI . Levels of Immunoglobulin E in Disease . . . . . A . Levels in Serum . . . . . . . . . B Levels in Secretions . . . . . . . . . C . Factors Influencing Immunoglobulin E Level . . . VII . Detection of Antibody Activity in the Immunoglobulin E Class A . Red Cell-Linked Antigen-Antiglobulin Reaction . . B. The Radioallergosorbent Test . . . . . . C. Allergen Antibodies in Serum . . . . . . . VIII . Metabolism . . . . . . . . . . . . . . . . . . . IX Concluding Remarks . References . . . . . . . . . . .

.

.

.

.

.

.

1

. . . .

. . . .

2 2

. .

. . .

.

.

. . . .

.

.

.

.

.

.

.

. . . . . . . .

.

. . . . . . .

. .

4 15 19 19 20 28 29 29 32 35 35 43 44 45 45 46 48 49 51 51

I. Introduction

Immunoglobulin E ( I @ ) represents a mincjr but distinct class of proteins present in serum of man and higher primates and possibly also in the serum of other species. In healthy individuals. the upper range of concentration is usually below 1 pg./ml. The detection and quantitation of IgE require very sensitive methods. Immunoglobulin E is elevated 4-30 times normal in various diseases. among which atopic disorders and parasitic infestations appear to be the most prominent . Pathological amounts of IgE have also been found in the serum of patients with yE myeloma. The association of certain reaginic antibodies to a new class of im1

2

HANS BENNICH AND S. GUNNAR 0. JOHANSSON

munoglobulin was postulated by K. Ishizaka and co-workers ( 1966a,b). The discovery that the myeloma protein ND and its normal counterpart share antigenic characteristics with reaginic antibodies and, in addition, carry skin-fixing structures, which appear similar to those of reagins, opened new possibilities to study the immunological and structural features of immediate hypersensitive reaction. The aim of this paper is to summarize our present knowledge of the biological and structural properties of IgE and its occurrence in various body fluids in health and disease, To this end, particular emphasis has been given to the methodology of identification and quantitation and also to the problem of isolation and characterization of IgE. References to the massive literature on reagins will be made only when found to be relevant for the understanding of a particular problem, and no attempts have been made to portray the long history of reagins, since this has been so masterly done in previous reviews by several authors (see K. Ishizaka and Ishizaka, 1968a; Sehon and Gyenes, 1965; Stanworth, 1963). II. Isolation and Physicochemical Characteristics of Immunoglobulin E

As a result of the obvious difficulties encountered in the isolation of reasonably homogeneous samples of a protein, which, like IgE, represents only a minor serum constituent, the physicochemical characteristics given in this paper will refer mainly to the first described E myeloma protein, ND. However, there is sufficient experimental evidence now available to support the belief that the ND protein has its major biological, immunological, and physicochemical characteristics in common with the IgE present in normal serum (Bennich et al., 1968). OF MYELOMA PROTEIN ND A. IDENTIFICATION

Our first attempt in 1965 to isolate the atypieal myeloma protein ND was done by zone electrophoresis ( Johansson and Bennich, 19674. The M component migrated in the fast y region. The isolated fraction, containing 93% of the M component contaminated mainly with IgG, was used for the first immunization experiments in rabbits and for carbohydrate analysis. The latter indicated that the M-component contained about 10%of total carbohydrate-a result suggesting a possible relationship of ND to IgA or IgM. Gel filtration experiments on serum ND on calibrated columns of Sephadex G-150 gave results in the same direction; indications that the M component distributed within the same elution volume as monomeric or 7 S IgA initiated a direct comparison of a monomeric A myeloma protein and protein ND. Both proteins were isolated from serum by precipitation with sodium sulfate and subsequently purified by recycling chromatography on Sephadex G-150 to

HUMAN IMhlUNOGLOBULIN E

3

eliminate contaminating IgG. The purified A and ND proteins were added to a solution of monomeric normal IgG and thc mixture was analyzed on a calibrated column of Sephadex G-200. The distribution of IgA and protein ND coincided completely as determined by quantitative immunological analysis, and the elution volume was significantly smaller than that of IgG. In contrast to these results, ultracentrifugal analyses indicated a significantly difference for IgA and protein ND, the sedimentation constant values ( so! ) were 6.5 and 7.9, respectively. Molecular weight determinations gave a value of about 139,000 for IgA and 196,000 for protein ND using a partial specific volume of 0.713 for both proteins. By reduction of protein ND with niercaptan followed by dissociation in acid, about 20% of the protein moiety could be rccovered as chains. The remaining 80%constituted a single carbohydratecontaining component with a characteristic electrophoretic mobility in starch gel electrophoresis in acid urea. This major constituent was regarded as representing the heavy chain of an atypical immunoglobulin. The problem of preparing class-specific antisera to IgE( N D ) was not solved until fragments of ND protein were isolated (see Section II1,B). Hereby it also became possible to develop the radioimniunosorbeiit test (RIST) described in Section IV and the radioallergosorbent test (RAST) described in Section VII. By using the RIST, a counterpart to ND was found in nornial serum. The concentration in healthy individuals are usually found to be extremely low as will be further discussed in Section V. However, by chance the serum from one of the apparently healthy blood donors included in the first series of experiments was found to contain a significantly highcr level of IgE( N D ) than the main level of the test group. The donor was subsequently found to have a previously clinically undiagnosed hypersensitivity to dog dander, a finding which initiated a study of the level of I g E ( N D ) in patients suffering from asthma and hay fever, as will be further discussed in Section VI. The significantly higher level of IgE( N D ) found in cases of extrinsic asthma strongly suggested a relation to reaginic antibodies as did the presence of allergen antibodies of IgE class in these patients. In 1966, K. Ishizaka et al. (1966a), from their studies on nntiragweed antibodies in reagin-containing fractions of atopic sera, suggestcd the prcsence of a unique immunoglobulin as a carrier of reaginic activity. The specific activity was found in the y, region by radioinimunoelectrophoresis and the protein was tentativcly designatecl YE-globulin. An exchange of antisera between Denver and Uppsala was made in March 1967 and, by comparatively antigenic analyses of myeloma protein ND and YE-globulin, direct imniunological evidence was obtained that ,lL.

4

HANS BENNICH AND S. GUNNAR 0. JOHANSSON

the two proteins are structurally related (Bennich et al., 1969b). Furthermore, these results gave direct support both for the hypothesis of a unique imniunogolobulin as a carrier of reaginic activity and for a relationship between reaginic antibodies and elevated levels of IgE ( ND ) in allergic serum. In addition, a biologicostructural relationship to reaginic antibodies was indicated by a specific inhibition of the Prausnitz-Kiistner reaction given by serum ND and purified protein ND at high dilutions ( Stanworth et aZ., 1967). B. ISOLATION 1. Salt Precipitation The recovery of IgE( ND) and IgG from serum precipitated with sodium and ammonium sulfate as followed by the single radial immunodiffusion technique of Mancini et al. (1965). A quantitative recovery of ND protein was obtained by precipitation with 40% saturated ammonium sulfate ( p H 7 ) or with anhydrous sodium sulfate (18 gm./100 ml. serum) at pH 7.5 and 25°C. These results are in agreement with those reported for reagins by Augustin and Hayward (1960) and by Sehon (1960). At 35% saturated ammonium sulfate, about 75%of the IgG was recovered as compared to 30%for IgE( ND). By adding 16 gm. of sodium sulfate to 100 ml. serum, IgG is quantitatively recovered but only 60%of I g E ( N D ) . The solubility properties of purified E myeloma protein were in close agreement to that of the M component in serum. Isolation of the latter by precipitation with salt was not found to alter its biological, immunological, and physicochemical properties ( Bennich, 1968). 2. Electrophoresis Protein ND migrates in the fast y to slow ,8 region in agar immunoelectrophoresis at p H 8.6. Reagins have been similarly characterized, and K. Ishizaka et a2. (1966a)' described the purified fraction of antiragweed antibodies, yE-globulin, to have yl mobility. Figure 1 illustrates the distribution of IgE relative to the other immunoglobulins in two sera from nonallergic individuals-one lacking IgA and IgD-after starch block electrophoresis for 24 hours at p H 8.6.

3. Chromatography Fractionation of serum ND by column chromatography on diethylaminoethyl ( DEAE )-Sephadex A-50 indicated that the M component distributed as a single peak, which by means of gradient elution (trisHC1 gradient from 0.1 to 1M , pH 8.0) was separated from other

5

H U M A N IMMUNOGLOBULIN E

1

I i I 1 I IDi

211011

1

I

'

I 15 -

5-

21-

3

M

-d E

21-

5

10

15

20

25

FIG.1. Electrophoretic distribution of immunoglobulins A, D, G, M, and E in two normal sera of which one (right-hand figure; sample G. S . ) was lacking IgA and IgU. Starch block electrophoresis ( 50 X 30 X 1 cni.), barbital buffer ( I = O . l ) , pH 8.6, 3 5 0 4 0 0 V., 100-110 mA., 20-24 hours, 4OC. Immunoglobulins A, D, G, and M were determined by single radial innnunodiffusion (Mancini et al., 1965) and IgE by radioimmunosorbent test. The concentrations of the immunoglobulins in the normal serum (left-hand figure) were A 1.1 mg./ml.; D 0.05 mg./ml.; G 10.5 mg./ml.; X I 0.49 mg./ml.; and E 510 ng./ml. In sample G. S.: A 0.01 mg./ml.; D 0.01 mg./mI.; G 12.8 mg./mI.; M 1.20 mg./mI.; and E 210 ng./rnI.


2

4 0.5

m a 0

v)

m

a 25

50

75

100

125

150

175

150

175

FRACTION NUMBER 0 . 6 ml vol.) 2.0

u E

I. 5

la

f 0

%

1.0

> 0 2

a

m

a

0

v)

0.5

m

a

25

50

75

100

125

FRACTION NUMBER 0 . 6 ml wol.)

FIG. 9. Gel filtration on Sephadex G-150 of serial samples of IgE(ND) undergoing papain digestion. Ultraviolet absorption; 280 nm. (line of black circles); hexose (line of open circles). The peaks were identified as IgE ( A ) , 7s fragment ( A ) , Fc (O), Fab, containing carbohydrate ( V ), Fab ( V), Fc" ( @ ) , and A-chain fragment, Ch ( 0).

fragment, designated Fc:, was found to be antigenically related to Fc, as will be discussed further below. Fragment Fc, was defined as the fragment in the digest that does not contain light chains but carries antigenic determinants, characteristic for the IgE class. In immunoelectrophoresis, Fc, occupies a position just anodal relative to intact IgE( ND), and by gel filtration on Sephadex G-150the fragment appears similar in size to that of F( ab') 2 y . Ultracentrifugation analysis indicated that Fc, has a molecular weight of about 98,000

TABLE 111 IMMUNOLOGICAL AND PHYSICOCHEMICAL PROPERTIES OF ENZYMATIC FRAGMENTS OF IMMUNOGLOBULIN E (ND)a ~

~~~~~

Tryptic fragm. Properties Molecular weightb Sedimentation const! ( s : , , ~ Carbohydrate

Papain fragm.

Peptic fragm.

Native IgE (ND)

75

Fc

Fab

Fab

Fc"

CX

7 s

F(ab)2 (t)

190,000

n.d.

95,000

50,000

40,000

38,000

25,500

n.d.

103,000 40,000 140,000 30,000

8.2S

7S

5.1 S

n.d.

n.d.

n.d.

n.d.

7 s

n.d.

6.75

6.75

n.d.

11.7

+

18.5

+

-

20-23

-

+

8

12-14

8.5

2.4

~

Fc"(t)

F(ab')2

pFc

(%)

Antigenic determinants" Epsilon, D.0 DJ D2 Light chain A Inhibitory P-K P-Kk activityd on (loo) (>loo) (10-20) (>40)

From Bennich and Johansson (1971); n.d. = not determined. Calculated on basis of r' = 0.713. D.0-idiotypic determinants; D.1 and D.2-determinants in the Fc portion of From Stanworth et al. (1968, 1970).

e

chains.

+-

+ +-

-

-

-

(+)

-

P-K n.d.

PCA(>28)

2

i 9

L(

PCA(>40)

F

HUMAN IMMUNOGLOBULIN E

23

(B = 0.72) and a sedimentation constant (s:,) of 5.1 S. Maximum yields of Fc, (about 30%)were obtained by 3 hours of digestion at p H 7 and 37OC using an enzyme to substrate ratio of 1:120 in the presence of cysteine (10 mM). Since the fragment represents an intermediary product, which gradually is cleaved to produce Fc: and smaller fragments including characteristic glycopeptides, isolated Fc, shows a considerable degree of microheterogeneity. Carbohydrate analysis indicates the presence of 14 to 17%carbohydrate, and starch gel analysis of reduced samples indicates the presence of one major and two minor components, which all three have similar mobilities. According to the definition, Fc, is antigenically indistinguishable from IgE ( ND ) using an antiserum specific to the latter. Furthermore, Fc, is the only fragment produced by papain that inhibits the P-K reaction in human skin (Stanworth et al., 1968) and PCA reaction in monkeys (Stanworth et al., 1970). Fragment Fc” represents an enzymatically resistant portion of the amino terminal part of Fc. It gives strong precipitin reactions with antisera specific for Fe, but is antigenically deficient as compared to the latter fragment; Fc” is a potent immunogen in rabbits. Antiserum to Fc” gives strong precipitin reactions with Fc, and F ( ab’),, distinct but slowly developing reactions with IgE( ND), but no visible reactions with partially reduced, alkylated E chains. The carbohydrate content of Fee" has been estimated to be about 20%and sedimentation equilibrium studies indicate that the fragment has a molecular weight of about 38,000 (7= 0.713; protein concentration = 0.5 mg./ml. ) . In agar immunoelectrophoresis (pH 8.6), Fc; is easily identified by its very fast anodal migration. In addition to Fc and Fc” fragments, two Fab fragments have been identified which differ in carbohydrate content. However, the yields of these fragments are very low due to their rapid breakdown into smaller fragments. The A chain of E myeloma ND appears to be very susceptible to attack by papain. The material in the CA peak (Fig. 9) reacted immunologically like A chain. However, both gel filtration data and starch gel analysis indicated that it constituted the constant portion of A chain; these results are in agreement with those obtained by enzymatic studies on Bence-Jones protein ND ( BerggBrd, 1970). 2. Tryptic Fragments

Trypsin splits IgE( ND) into a 7 s fragment, which by prolonged digestion yields F( ab),( t ) and Fc”(t ) and smaller peptides, but no Fc( t ). The chemical and imniunological properties of tryptic fragments are summarized in Fig. 10 and Table 111. By short-time digestion (less

24

> 0

C

m

n L

0

In

n

a

E l u t i o n v o l u m e , Ve : Vt

B

5

z

0

3

C

a 9 L

0 y1

n

a

1

50

150

250

3 50

Fraction ( 4 m l )

FIG. 10. Top: gel filtration on Sephadex G-150 of a tryptic digest (1 6 hours) of I g E ( N D ) . The peaks were identified as IgE ( A ) , 7 s fragment ( B ) , and F( ab)?(t ) ( C ) . Bottom: chromatography (diethylaminoethyl-Sephadex A-50) of a tryptic digest of IgE( ND). Peaks identified as shown in top figure. Peak D contained in addition to trypsin inhibitor, also Fc”( t).

HUMAN IMMUNOGLOBULIN E

25

than 60 minutes), a 7 S fragment is the main product, which is essentially indistinguishable from the F( ab’), fragment produced by pepsin, However, analysis of reduced samples of 7 s fragment indicates a difference as regards constituent heavy-chain fragments. The tryptic 7 s fragment was found to inhibit PCA reactions in monkey skin. However, these findings must be considered with caution since the inhibitory activity appeared weak on a molar basis and contamination with intact IgE( ND)-less than 5%contamination in purified 7 S preparations would be sufficient-could not be excluded. By further digestion, a smaller but immunologically related fragment is produced. The molecular weight of this fragment, designated F ( ab) ( t ), is about 103,000 ( = 0.713; protein concentration 0.2 mg./ml. ) . The F( ab),( t ) fragment is antigenically deficient as compared to Fc, produced by papain and lacks PCA inhibitory activity. By prolonged digestion a resistant fragment of about 40,000 molecular weight is produced in increasing yields which was designated Fc”(t) since it is similar to papain Fc”. 3. Peptic Fragments

The physiochemical and immunological properties of peptic fragments are shown in Fig. 11 and Table 111. Digestion of IgE(ND) by pepsin at pH 4.5 produces two fragments, both rich in carbohydrate. The inajor fragment, carrying both h antigenic sites and some of the antigenic determinants characteristic for c chains, was assumed to represent F( ab’ ) fragment ( Bennich and Johansson, 1967). However, the yield was surprisingly high-about 75% as calculated on UV basisand the carbohydrate content represented about two-thirds of that of IgE ( ND). The remaining one-third of carbohydrate was recovered together with a minor fragment, designated pFc, which constituted about 15%of the native protein. About 10%of the digest was recovered as small immunologically inactive peptide fragments completely devoid of any carbohydrate. Ultracentrifugal analysis of F( ab’ ) fragment indicated a molecular weight of about 140,000 + 4000 and a sedimentation constant ( ) of 6.7 S . After reduction and alkylation the fragment dissociates in two components, which were isolated by gel filtration in acid buffer. The minor component was identified as X chains and, accordingly, the major, carbohydrate-containing, component was assumed to constitute F d fragment. The latter has a molecular weight of about 45,000 as found by sedimentation equilibrium studies in 6 M guanidine of completely reduced and alkylated fragments ( Dorrington and Bennich, 1971). Partially reduced alkylated F( ab‘)? fragments dissociate at neutral pH to give Fab’ which

I

5

3 > 0 C

m

n L

0 U

n

U

1

E I u t i o n v o l urn 8 , V, : Vt

5

>

0

3

C

m 0 L

0 U

n 4 -I

1

Fraction ( 4 m l )

FIG.11. Top: gel filtration on Sephadex G-150of a peptic digest (20 minutes) of IgE(ND). The peaks were identified as F(ab'): ( A ) and pFc ( B ) . Immunoglobulin E ( N D ) is eluted by a volume of 0.45 V t . Bottom: chromatography (diethylaminoethyl-Sephadex A-50) of a peptic digest ( 8 hours) of IgE(ND). Peaks identified as in top figure.

HUMAN IMMUNOGLOBULIN E

27

indicates that inter-heavy-chain disulfides have cleaved. Fragment F ( ab’) comes off Sephadex G-150 columns in a volume fraction similar to that characteristic for human IgG. By chromatography on DEAESephadex A-50, the fragment is recovered in fractions of 0.2 to 0.3 M tris-HC1, pH S.0 (see Fig. 11).In immunoelectroplioresis (agar pH &6 ), F(ab’), is located on the cathodal side of IgE( N D ) similar to Fab or Fab’ fragments produced by other proteolytic enzymes and similar also to the behavior of 11 S IgE fragments isolated from serum (see Section I1,B). Fragment F( ab’) is precipitated by antiserum specific for Fc ~, it lacks, however, some of the characteristic antigenic sites common to Fc and IgE( ND), but will give a reaction of identity with Fc” [and F c ” ( t ) ] using antiserum specific either for Fc” or for Fc. The cleavage of IgE( ND) by pepsin at pH 4.5 and 37°C is a very rapid process; within less than 15 minutes more than 90%of the substrate is cleaved to yield F( ab’) and pFc fragments and small peptides. The immunological and physicochemical properties of F( ab’ ) do not change by prolonged digestion up to about 24 hours. The pFc fragment, which is enzymatically labile, represents a portion of the carboxy terminal part of the E chains that carries some of the antigenic determinants characteristic of Fc. The molecular weight of pFc fragment was determined to be about 30,000 ( V = 0.713; protein concentration 0.5 mg./ml.) by sedimentation equilibrium experiments. The carbohydrate content is high, about 24%,which corresponds to about one-third of the total carbohydrate of native IgE( ND), By adding the carbohydrate data for F(ab’), calculated on the basis of a molecular weight of 135,000, the sum of carbohydrate residues will be in close agreement with the carbohydrate data for IgE( N D ) (Bennich and Clamp, 1971). Attempts to inhibit PCA reactions with pFc fragments have so far been unsuccessful. In summary, fragmentation studies of IgE ( N D ) have shown that antigenic determinants ( D ) , specific to the c chain and located within the Fc fragment, can be separated in two groups, in the following referred to as D,1 and D,2. Fragments F (ab’)? and Fc share the antigenic determinants, D J , which arc localized within the amino terminal portion of the Fc fragment. This part of thc E chain, which represents a glycopeptide consisting of about 120 amino acid residues, is characterized by a relative resistance to papain, pepsin, or trypsin, and, even by prolongc~ldigestion, a covalently linked dimcr of about 40,000 molecular weight [Fc” or Fc”(t ) J having D,1 specificity can be recovered. Amino acid data and reduction experiments indicate that Fc” represents a “domain” containing four half-cystinyl residues of which two are engaged in interchain bonds. In contrast, structures carrying

28

HANS BENNICH AND S. GUNNAR 0. JOHANSSON

D,2 specificity are readily cleaved to produce inactive fragments. No fragments have been identified to carry only D,2 specificity with the exception of pFc which represents a noncovalently linked glycopeptide from the carboxy terminal part of Fc. Separate regions carrying D,1 and D,2 specificity can be shown to be present also on IgE from normal and atopic sera and from secretions (Bennich and Johansson, 1971). The inhibitory activity of IgE( ND) on reagin-induced PCA in baboons (or P-K in humans) has so far been recovered only in the Fc fragment, which carries both D,1 and D,2 specificity. However, the absence of inhibitory activity in other fragments does not justify speculations on the further localization or nature of the skin-fixing sites of c chains, but rather directs the attention to a weakness of indirect methods such as should be complemented by techniques for a direct study of the nature of binding of IgE and its fragments to cell surfaces. IV. Methods for Determination

The best way to determine IgE is to use immunochemical methods based upon the reaction between IgE and antibodies specific for the TABLE I V COMPARISON OF RADIOIMMUNOCHEMICAL METHODSFOR ESTIMATION OF IMMUNOGLOBULIN E Criteria

RISTa

Sensitivity (IgE estimation) lteproducibili ty Capacity; samples per day and technician Stabilky of reagents Reagents needed, antibody

1 ng./ml. 5-1Oa/, 100

antigen

Results obtained within Special equipment required

0

Catt “sandwich”b 1 ng./ml. 5-1070

100

Very good Very good Very small Very small quantity quantity specific specific Very little, pure Rather much, pure or crude 8-24 hours Scintillation detector; centrifuge for washing

8-24 houm Scintillation detector

RIST-radioimmunosorbent test. From Salmon et al. (1969); Johansson and Bennich (1970). RSRD-radioimmune single radial ditrusion. From Rowe (1969).

RSRD 40 ng./ml. 10-200/, 50

Very good Very small quantity specific Serum with known IgE concentration 5-10 days None

HUMAN IMMUNOGLOBULIN E

29

class-related antigenic determinants of IgE. Gel diffusion methods, such as single radial diffusion in gel (Mancini et al., 1965) are handicapped by low sensitivity; radioimmune assays have proven to be superior. Only a few radioimmune methods have been used for IgE estimation. RIST (Wide and Porath, 1966; Johansson et al., 1968a) and a ''sandwich" modification (Salmon et at., 1969) of the solid phase method described by Catt et al. (Catt and Tregear, 1967; Catt et al., 1967) and the double antibody technique (Gleich et al., 1971) seem to be useful. A two-step modification of the Mancini method was described by Rowe (1969) where the introduction of isotope-labeled antiglobulin antiserum enabled the detection of IgE levels as low as 50-100 ng./ml. In Table IV some criteria, such as sensitivity and precision of the methods, are compared. V. levels of Immunoglobulin E in Healthy Individuals

A.

LEVELSIN SERUM

The dominating immunoglobulin in cord serum is IgG. Only very low concentrations of IgM, about 10%of adult mean level (Franklin and Kunkel, 1958; Johansson and Berg, 1967) can be found. Immunoglobulin A and D are usually below the limit of detection for gel precipitation methods (Johansson and Berg, 1967); IgE can be detected in cord serum with the use of RIST. In a study of samples from healthy newborns, a mean IgE concentration of 36 to 38 ng./ml. was found with a range of 16.0 to 97.5 ng./ml. (Johansson and Bennich, 196%; Johansson, 1968a). This level corresponds to about 15%of the adult level. Since the IgE values did not distribute in a Gaussian manner, before calculations all values were changed to logarithms to the base 10. The confidence limits (mean value +2 S.D.) for the IgE in the cord serum study was 12.9-102 ng./ml. Sera were also collected from 14 anamnestically allergic mothers and from their respective newborns (cord serum). No correlation was found ( T = 0.002) between the IgE concentration of the mothers and their respective newborn (Johansson, 1968a). This seems to indicate that the IgE present in cord serum consists mostly of material synthesized de novo by the fetus. The quantitation of such low IgE concentrations is open to some question due to the technical problems involved. Recent studies seem to indicate that the IgE concentration in cord serum is even lower than that reported. In Table V the IgE concentration in cord serum is compared with the IgE levels in children and adults. The development of the immunoglobulin levels during childhood have been studied by many authors (Claman and Merrill, 1964; Stiehm and Fudenberg, 1966; Collins-Williams et al., 1967; Johansson and Berg,

30

HANS BENNICH AND S. GUNNAR 0 . JOHANSSON

TABLE V SERUMIMMUNOQUJBULIN E LEVELSIN HEALTHY CHILDREN A N D ADULTS. IgE levels (ng./ml.)

Serum Cord serum Children If-44 months 4+-9 months 9 months-3 years 3-5 years 6-10 years 11-15 years Adults (I

Confidence limits (96% interval)

Geometric mean

Range

36. 3

12.9-102

16.0-97.5

60. 6 75.7 114 158 190 246 248

43.5-84.5 24.7-233 29.0-450 45.3-528 55.6-648 71.9-838 61.4-1000

50.0-86.0 24.0-223 49.5-540 62.0-308 63.0-535 54.0-840 66.0-1830

From Johansson (1968a) and Berg and Johansson (1969b).

1967). In Fig. 12 the development patterns for IgG, IgA, IgM, IgD, and IgE are summarized. The striking difference between the slow rise of IgA and the fast increase of IgM is well known. The pattern of IgD in the first year is not yet known but will be easy to investigate by the new highly sensitive methods. In a first study of the IgE development in 50 healthy children, 6 weeks to 5 years of age, a rather slow increase with age was found (Johansson, 1968a). Since the regression

/

mg /ml

mg I00 ml Partus

I

............. gM9' (000 ..................l g M d

4

I

Week of gestation

Months

500

Years

FIG. 12. The development during childhood of the serum concentrations of the five immunoglobulins. Only the mean levels are given. (Data from Johansson and Berg, 1967; Berg, 1969; Berg and Johansson, 1969b; by permission of Kabi, Stockholm. )

HUMAN IMMUNOGLOBULIN E

31

line satisfying all samples gave a level at 6 weeks of age that was somewhat higher than the mean of cord sera, a pattern for IgE similar to that of IgM was proposed. However, further studies (Berg and Johansson, 1969b) of 138 randomly sampled healthy children gave a distribution pattern more like that of IgA. When 10 children were followed with repeated samples during their first year of life (Berg and Johansson, 1969b), a very slow increase of IgE concentrations with age was found. This is in good agreement with IgA but is in sharp contrast to what has been found for IgM (Johansson and Berg, 1967; Berg, 1968). Even in premature newborns, a very fast increase in IgM has been reported (Berg and Johansson, 1967b; Berg, 1969). The difference in IgG and IgM concentrations which was found in boys and girls could not be shown for IgE (Berg and Johansson, 1969b). This is of particular interest since it is well known that of children suffering from asthma, there are twice as many boys as girls. With aging in adults, only small changes in the immunoglobulin concentrations (Johansson et al., 1968b) are seen, and this seems also to be true for IgE (Johansson, 1968a; Johansson et al., 1968b). The difference between men and women which has been reported for IgM and perhaps also for IgD (Johansson et al., 1968b) could not be found with IgE. The decrease of allergic disorders which occurs with increasing age does not seem to be accompanied by a decrease in IgE concentrations. One complication in the studies of the normal distribution of IgE is in the selection of healthy, nonallergic individuals. For practical reasons, we had to select our cases from interviews by a clinical allergologist. No allergological investigation such as skin or provocation tests were made. By this procedure some individuals with subclinical allergy might have been included. Supporting this idea are the findings of a tendency for subnormal IgE levels in patients with asthmatic symptoms but with negative findings on allergological investigation (Johansson, 1967; Berg and Johansson, 1969a). In a study (Berg and Johansson, 1969a) of children with asthma (from Dr. Aas, Olso), it was found that the 28 children with negative findings on clinical investigation had a mean IgE level of 113%of mean for the age. Twenty of these had a positive histamine provocation test, indicating hypersensitive bronchial mucosa which is found in high frequency among allergic individuals. Their mean IgE level was 134%.The other 8 had normal bronchial histamine sensitivity and were, therefore, so far as was possible to evaluate at the time, nonallergic. Their mean IgE level was only 62%. The influence of genetic factors on the IgE Ievels has been studied (Rowe et al., 1971) in a twin population. One hundred and twenty-eight twin pairs were studied, of which fifty-nine were monozygotic (Rowe

32

HANS BENNICH AND S. GUNNAR 0. JOHANSSON

et al., 1968a). For monozygotic men below 20 years of age, a significant intrapair variance was found, This was the only result that indicated a genetic influence on the IgE levels. Obviously the environmental factors are of importance for the development of the IgE concentrations. These results fit very well with the meaning of atopy: an increased tendency, influenced by hereditary factors, to develop disorders based upon immediate-type hypersensitivity reactions. The relation between the concentration of IgE and IgA in an individual was investigated in the twin study. The absence of correlation between IgE and IgA would seem to indicate that there is no firm relationship for the production of these two immunoglobulins.

B. LEVELS IN SECRETIONS The reagin-mediated immunological reactions of asthma and hay fever take place in or in the immediate neighborhood of mucose membranes. It could very well be that the circulating IgE represents an excess of reagin from the affected organ, the lung, or the nose. The information about IgE in secretions is limited (Johansson et d., 1971a). The IgE concentrations in saliva from healthy individuals have been estimated by RIST in a preliminary study (Turner and Johansson, 1971). Very low concentrations were found ranging between 1 to 10 ng./ml. No significant difference in absolute concentrations could be found between whole saliva and parotid secretion. The IgE/IgA (serum comIgA was used as a standard) ratio for whole saliva was 5.7 X pared to 13 x for parotid secretion. The same ratio for serum was 0.1 x The tendency for relatively lower IgE concentrations in the whole saliva might be attributed to addition of gingival pocket secretion which is known to have an immunoglobulin composition very similar to that of serum (Brandtzaeg, 1965). The absolute IgE levels in gingival pocket fluid in healthy adults and patients with parodontiasis seem to be the same as in the serum (Killander et al., 1971). A few samples of nasal washings from healthy individuals have been studied. Very low levels of IgE were found, the absolute concentrations ranging from 3 to 30 ng./ml. The IgE levels have also been studied in bronchial washings from 20 patients. Immunoglobulin E could not be detected in 12 of the unconcentrated secretions. The mean IgE concentrations in the other 8 samples was 5.4 ng./ml. with a range of 1.8 to 12.0 ng./ml. (Deuschl and Johansson, 1970). It is obvious that it is possible to detect IgE in nasal and bronchial secretions of healthy individuals by RIST. Further studies are now in progress to estimate to what proportion the amount of IgE can be attributed to local production. Nasal polyp fluid from allergic patients is known to contain very high

33

HUMAN IMMUNOGLOBULIN E

concentrations of reagins ( Berdal, 1954). Relatively high levels of IgE were also found in individuals without any signs or symptoms of atopic allergy (Donovan et al., 1970). However, since nasal polyps cannot be regarded as a normal status, this condition will be discussed in detail in Section VI,B. The secretion containing the highest immunoglobulin concentrations, and predominately IgA, is colostrum. The IgA concentration can reach a few grams per 100 ml. The IgE levels in colostrum have been studied in the first week of lactation. Very high concentrations were found, in some mothers more than 20 times higher than that of serum. In Fig. 13 is shown the development of the IgE levels in colostrum during the first week of lactation. Two different patterns are shown. In one case the highest levels were found about day 1-2 after delivery and, thereafter, they decreased; in the other case, a steady increase was seen up to day 5-6. When the IgE/IgG ratio in the colostrum was calculated, a very fast rise was found in both patients; the slopes of the two curves were almost parallel. These findings can be explained on the basis of 1oooc

1000

P

LOO0

LOO

2000

200

1000

100

-2 LOO

10

200

10

100

10

* X

0

C

2 -m 0

-

e'

0 PARTUS

1

2

3

4 5 Days

6

7

8

FIG. 13. The development of IgE levels in Iiunian colostrum from two women, ( 0) and ,).( respectively, during the first days of lactation compared with serum IgE levels, and ( @ ) , respectively. Imniunoglobulin E levels are given in absolute concentrations (-) and as a IgE/IgG ratio in colostrum ( - - - - ) .

(a)

34

HANS BENNICH AND S. GUNNAR 0. JOHANSSON

selective transport of serum IgE or by local synthesis. Since there seems to be no correlation between the concentration of IgE in serum and colostrum the hypothesis of local production is likely (Johansson et al., 1971a). Preliminary studies on the molecular size have shown that the major part of clostrum IgE comes off a calibrated Sephadex G-150 column in the same volume as serum IgE (Fig. 14). The immunoglobulin concentration, with special reference to IgE, has been studied in urine from 16 healthy individuals (Turner et al., 1970). The immunoglobulins were simultaneously quantitated in serum and urine, and clearance data were used to estimate the amount of protein locally produced. The results indicate a slight local production of IgG and IgA. A very high clearance of IgE was found, which is thought to represent local production. As much as 100 times more IgE was found than would have been expected from the clearance data. The amounts of IgG and IgA produced locally in percentage of total urinary IgG and IgA varied from 9 to 92%. The same percentage for IgE showed less variation ranging from 62 to 100%.The high urine IgE concentrations can also be explained on the basis of degradation of IgE in the urinary tract. However, so far the only in vitro produced frag-

L

0 u)

n

a

0.20

0.32

0.44

0.56

0.68

E l u t i o n v o t u m e , v,:

0.80

0.92

1.04

vt

FIG. 14. Gel filtration of colostrum ( 6 ml.) from a nonallergic woman on a calibrated column (3.2 X 92 cm.) of Sephadex G-150. Absorbancy, 280 nm. (0); colostral IgE, as determined by radioimmunosorbent distribution of a purified sample of IgE(ND) (17).

(a);

HUMAN IMMUNOGLOBULIN E

35

ments containing E determinants are the F( ab’)? and the Fc fragments, both having a molecular weight larger than that of albumin (Section 111). The lack of correlation with serum lcvels on patients with atopic diseases and various renal disorders also is in favor of a local synthesis. It, therefore, seems very likely that the IgE in urine is produced locally somewhere in the urinary tract. The function of such IgE is yet unknown although some beneficial function is perhaps more probable than a local urinary reagin-mediated disorder. The findings of IgE in several secretions and the data indicating that this is the result of a local production are very interesting. Fluorescent studies using specific antisera to IgE have shown a localization of IgE-staining plasma cells to mucosal membranes of the respiratory and gastrointestinal tracts ( Tada and Ishizaka, 1970). Further studies are of interest to evaluate the importance of local IgE in relation to serum IgE and its role in disease. VI. levels of Immunoglobulin E in Disease

A. LEVELSIN SERUM 1. Atopic Diseases In a preliminary study of patients suffering from asthma and hay fever (Johansson and Bennich, 1967b), some cases with highly increased serum levels were found. The mean IgE value in a group consisting of 31 cases with proven allergy was 1191 ng./ml. with a range of 133 to 5850 ng./ml. These values should be compared with mean IgE of 2,48 ng./ml. in healthy individuals. The highest concentration observed in this study, 5850 ng./ml., is about 20 times higher than the normal mean. In order to investigate the I g E distribution in asthma, a group of adult patients with asthma was analyzed (Johansson, 1967). The study consisted of 38 cases which, according to the allergological investigation, were divided into two groups. The patients were tested by intradermal tests using fifteen commercial allergens including four pollen mixtures and nine animal dandruffs, mold mixture, and house dust. Positive skin reactions were verified by a bronchial provocation test. All patients with positive skin and provocation tests to at least one allergen were brought together in a group called “allergic asthma.” The other group of patients was called “nonallergic asthma.” No difference was found for the concentrations of IgG, IgA, IgM, and IgD between the allergic and nonallergic group. This is in contrast to the findings of Kohler and Farr (1967) who reported high concentrations of IgD in allergic diseases.

36

HANS BENNICH AND S . GUNNAR 0. JOHANSSON

The mean IgE level for the whole study was 828 ng./ml. which is about 3 times the normal mean level. However, the allergic group had a mean of 1589 ng./ml. compared to 275 ng/ml. in the nonallergic group. In the allergic group, 8 of the 16 patients had pathologically high levels (more than 1000 ng./ml.). Immunoglobulin E levels were also studied in children with asthma and hay fever (Berg and Johansson, 1969a). Samples were analyzed from 94 untreated children (70 boys and 24 girls) which all had a positive allergological investigation including provocation tests. As can be seen in Table VI, 13 children had only symptoms of asthma. Their IgE mean level was 563%of normal mean for their age. The 22 children with rhinoconjunctivitis as the predominant symptom had a mean IgE level of 297%. One explanation for this difference could be the more severe and prolonged symptoms that usually occur in asthma. Thus it could be shown that children with perennial symptoms of asthma had a higher mean IgE level (711%)compared to the children with symptoms mainly during pollination season (230%).Of 22 children with hay fever, only 8 had elevated levels, and the mean IgE value for the group was 297%.The influence of allergenic stimulation on the IgE concentrations was clearly shown when the IgE levels were followed in untreated children during a pollination season. The mean level before season was 12652, during season 286%,and after season 242%. TABLE VI IMMUNOGLOBULIN E LEVELSI N CHILDREN WITH ATOPIC DISEASES~

Diagnosis Bronchial asthma Hay fever Bronchial asthma and hay fever Bronchial asthma and atopic eczema Hay fever and atopic eczema Negative allergologic investigation

No. of children

No. of children with high levels*

Mean IgE concentrationc

13 22 2s

10 8 15

563 297 350

S

7

744

4

2

599

7

1

111

Data from Berg and Johansson (1969a). Immuiioglobulin E concentration higher than mean $ 2 S.D. for the age. 111 percent of the predicted arithmetic mean value for healthy children of the same age.

HUMAN IMhfUNOGLOBULIN E

37

Different types of allergens might be more-or-less potent in their ability to stimulate the IgE production. Thus, dust and mold allergens have been regarded as weak allergens by the allergologists, whereas common animal dandruff and pollen allergen have been regarded as strong allergens. In a study (Berg and Johansson, 1969a) of children with asthma (from Dr. Aas, Oslo), it was found that 11 children with positive skin and provocation tests for dust and/or mold had a mean IgE level of only 104%of mean for their age compared to 338% for 42 children allergic to other common allergens. Atopic eczema (prurigo Besnier) is a disease the relationship of which to reagin-mediated disorders has been discussed. It is, therefore, of interest that patients with atopic dermatitis have raised IgE levels. In the study of allergic children (Berg and Johansson, 1969a) it was found that when the children had asthma and eczema or hay fever and eczema, the mean IgE level was 744 and 599%of mean for their age, respectively, compared to 563 and 297%in pure asthma and hay fever, respectively. Clear-cut changes were found when adult patients with pronounced atopic dermatitis were investigated (Juhlin et al., 1969). Studies of 28 patients showed a mean IgE value of 2733 ng./ml. which corresponds to about 11 times the normal mean. As high a value as 31,000 ng./ml. was found in 1 patient who in addition to his eczema also suffered from severe asthma due to animal and pollen hypersensitivity. Five patients (18%)had normal IgE levels, but this number increased with the number of less severe cases included (Ohman and Johansson, 1971). 2. lmmunobgical and lnfectious Diseases The Wiskott-Aldrich syndrome is characterized by increased susceptibility to infections, thrombocytopenia, and eczema ( Wiskott, 1937; Aldrich et al., 1954). Immunoglobulin E concentrations were measured in a study of 6 cases of Wiskott-Aldrich syndrome (Berglund et al., 1968). High levels were found in all 6 patients studied, but there was a remarkable change in the levels with time. Further studies of the children seem to indicate that the IgE level varies with the state of the eczema. When the eczema was pronounced the IgE value was high. Concentrations of IgE have been studied in immunological disorders other than the Wiskott-Aldrich syndrome. No significant changes were found in diseases such as rheumatoid arthritis, systemic lupus erythematosus, and ulcerative colitis. In the latter group, 49 patients were studied (Johansson et al., 1 9 7 1 ~ ) The . mean IgE value for the group was 334 ng./ml., but 3 of the patients had raised levels (1063, 2500, and 4300 ng./ ml. respectively) and 1 patient had a hypogammaglobulinemia

HANS BENNICH AND S. GUNNAR 0. JOHANSSON

38

TABLE VII SERUMIMMUNOGLOBULIN LEVELSIN PATIENTS WITH M COMPONENTS, HYPOQAMMAGLOBULINEMIA, A N D SELECTIVE IMMUNOGLOBULIN A DEFICIENCY Age (years)

Sex

Diagnosis

IgEa

IgGb

IgAb

IgMb

IgDb

65 64 69 74 59 65 63

m f f m m f m

B.H. G.E. S.L.e A.K.1 L.S. B.K. A.K. G.S. J.R. B.H.

6120 5000 280 1410 525 5000 1420 19 232y 262 Denham, S., 114, 120, 197 de Saussure, V. A., 13, 52 Desmyter, J.> 151*Ig7 de Sousa, M. A. B., 260, 264 Des Pres, R. M., 302(129), 312 Destombes, P., 238, 239, 242, 261, 262 Deuschl, H., 32, 52 de Vassal, F., 176, 202 Deverill, J., 62, 63, 68, 98, 99 De Weck, A. L., 272(9), 300(9), 308 Dharmendra, 214, 262 Diakitk, S., 248, 263 Dias da Silva, W., 278(53), 282(53), 283(53), 310 Dickenson, J. B., 165, 202 Dienes, L., 170, 185, 197

;

~

~

319

AUTHOR INDEX

Diengdoh, J. V., 136, 170, 197, 206 Dierks, R. E., 230, 235, 262, 265 Dixon, F. J., 138, 203, 272(12, 13), 273 (13), 274( 13), 288( 13), 303( 13, 131, 132), 305( 137), 308, 312 Dodd, R. Y.,177, 197 Doenhoff, M. J., 105, 118, 161, 187, 197 Dolovich, J., 301( 121), 312 Donaldson, V. H., 279( 63), 307( 143), 310, 313 Donovan, R., 33, 44, 52 Donsky, G., 291(105), 311 Dorreter, J. B., 146, 202 Dorrington, K. J., 16, 19, 25, 53 Dostrovsky, A., 239, 262 Douglas, A. P., 39, 52 Douglas, S. D., 105, 195 Draper, P., 215, 262 Dray, S., 68, 97, 127, 129, 136, 201, 206 Dreyer, W. J., 84, 97 Dubert, J. M., 94, 98 Dulbecco, R., 269( 4 ) , 308 Dunionde, D. C., 127, 136, 141, 144, 157, 160, 197, 201, 207, 242, 2.17, 248, 250, 251, 252, 254, 255, 256, 257, 258, 259, 262, 264 Dunan, S., 246, 249, 264, 265 Duplan, J. F., 94, 98 Dupuy, J. M., 154,197,203 Duskenski, L. A,, 301(121), 312 Dutton, R. W., 107, 160, 162, 164, 187, 188, 189, 191, 193, 197, 200, 202 Duveen, G. W., 44, 54 Duxbury, R. E., 248, 262 Dwyer, J. M., 161, 197 E

East, J., 260, 264 Edelhoch, H., 16, 17, 53 Edelnian, R., 166, 197 Ehrcnberg, A., 15, 53 Eichmann, K., 71, 79, 80, 97 Eilber, F. R., 176, 202 Ein, D., 84, 98 Einbinder, J. M., 278(58), 310 Eisen, H. N., 186, 200, 269( 4 ) , 308 Elkins, W.L., 174, 197 Elliot, E. V., 187, 196 Elliot, J. H., 176, 198

El-Shawi, N. N., 236, 265 Elves, M. W., 104,197 Engle, R. L., Jr., 62, 91, 98 Epstein, W., 91, 99 Eriksen, J., 63, 98 Ernback, S., 10, 51, 54 Escobar-Gutienes, A,, 232, 265 Evans, R., 175, 197 Evans, R. R., 307( 143), 313 Evans-Paz, Z., 237, 242, 262

F Fabris, N., 290(98), 311 Fahey, J. L., 39, 54, 274( 19), 309 Falk, R. E., 143, 145, 147, 197, 207 Fantes, K. H., 148, 149, 150, 197, Farah, F. S., 241, 263 Farr, R. S., 35, 54, 276(37), 309 Fasal, P., 230, 262 Fauve, R. M., 135, 197 Federlin, K., 165, 197 Feinman, L., 305( 135),312 Feinstein, D. I., 298( 112), 312 Feinstone, S. M., 127, 129, 197 Feizi, T., 71, 79, 80, 97 Feldman, J. D., 171, 184, 195, 197, Feldman, M., 117, 196 Fellner, M. J., 107, 198 Fermresi, R. W., 126, 129, 198 Ferri, R. G., 246, 262 Festenstein, H., 105, 118, 197, 260, Finnstrom, O., 37, 52 Finter, W. B., 149, 200 Firschein, I. L., 164, 199 Fischer, H., 114, 119, 178, 193 Fischer, O., 2.37, 263 Fish, A. J., 107, 198 Fishel, C. W., 290( 101), 311 Fisher, D. B., 105, 119, 198 Fishkin, B. G., 50, 54 Fishman, M., 191, 198 Fikgerald, P. H., 106, 203 Fitz-Herbert, M., 239, 242, 264 Flas, M. H., 176, 198 Foley, H. T., 229,261 Foner, A,, 244, 257, 261, 263, 266 Forbes, I. J., 105, 206 Ford, C. E., 104, 105,198 Forni, L., 165, 204 Foster, W. A., 239, 241, 244, 248,

200

203

263

263

320

AUTHOR INDEX

Foucard, T., 32, 34, 44, 48, 52, 53 Frame, M., 274( ZO), 309 Frangione, B., 274( 17), 309 Frank, H. A., 174, 201 Franklin, E. C . , 29, 53, 274( 17, 19), 309 Freedman, S. O., 107, 198 Friedman, R. M., 149,198 Fudenberg, H. H., 28, 29, 39, 53, 54, 55, 63, 83, 84, 85, 90, 93, 98, 99, 108, 124, 126, 144, 153, 183, 200, 205, 206, 236, 263, 299( 117), 312 Fulginiti, V. A., 259, 263

Gleich, G. J., 29, 53 Glynn, L. E., 129, 204, 284(86), 311 Godal, T., 213, 263 Godfrey, H. P., 108, 126, 129, 196, 198 Godwin, H. A., 229, 261 Goidl, E. A., 188, 189, 191, 200, 203 Gold, E. E., 129, 204 Goldberg, A. F., 199 Goldberg, B., 166, 170, 198 Goldie, J. H., 189, 198 Golub, E. S., 198 Good, R. A., 39, 40, 51, 52, 102, 103, 154, 179, 180, 196, 197, 202, 203, 204, 259, 262 Gordon, B., 239, 262 Gordon, F., 148, 198 Gordon, J., 110, 143, 198 Gotoff, S. P., 136, 201 Govaerts, A., 112, 198 Gowans, J. L., 104, 105, 171, 190, 198, 206, 207 Gowland, E., 154, 198 Graf, M., 304( 134), 312 Grangaud, J. P., 246, 263 Granger, G. A., 119, 137, 138, 139, 140, 141, 142, 143, 153, 173, 175, 198, 199, 201, 207 Granner, D., 104, 206 Grant, C . K.,’114, 120, 197 Graupner, K., 163, 193 Gray, D. F., 177, 199 Gray, H. K., 129,204 Greaves, M. F., 107, 119, 146, 161, 186, 189, 190, 195, 199,205 Green, J. A., 141, 150,199 Green, N. M., 67, 91 Gresser, I., 142, 151, 199 Grey, H. M., 50, 54, 63, 84, 88, 91, 93,

G Gaafar, S., 217, 218, 219, 263 Galindo, B., 127, 129, 198 Garbe, G., 282(82), 283(82), 311 Garcia-Giralt, E., 143, 198 Garnham, P. C . C., 246, 247, 249, 259, 263, 264 Garrido-Pinson, G. C., 298( 114), 312 Gaugas, J. M., 226, 227, 229, 263, 264 Gelehrter, T. D., 104, 206 Gelfand, E. W., 299(44b), 310 Cell, P. G. H., 58, 59, 63, 65, 71, 75, 87, 94, 97, 98, 106, 154, 182, 183, 186, 188, 194, 198, 203, 205, 268 ( 2 ) , 308 Gemetchew, T., 239, 241, 244, 248, 263 George, M., 123, 129, 198 Gerety, R. J., 126, 129, 198 Gery, I., 107, 194 Gettner, S., 258, 265 Gibson, T., 170, 198 Gigli, I., 278( 52), 310 Gilchrist, C., 227, 263 Gillette, R. W., 171, 198 Gillissen, G., 165, 198 97, 99 Gilman, A., 68,97 Gribik, M., 126, 129, 136, 205 Ginsberg, H. S., 269(4), 308 Griffith, D., 259, 264 Ginsburg, H., 117, 164, 198 Grynberg, N. F., 264 Girard, J.-P., 8, 53, 107, 198 CuiffrB, L., 246, 264 Githens, J. H., 259, 263 Guinto, R. S . , 227, 263 Gitlin, D., 259, 263 Gunders, A. E., 257, 263 Glade, P. R., 103, 111, 138, 143, 146, Gurner, B. W., 126, 153, 193 149, 194, 196, 198, 199, 200 Guthe, T., 272(9), 300(9), 308 Glasgow, L. A., 150, 198 Gutman, R, A., 43,55 Glasscock, R. J., 303( 131), 312 Guttman, R. D., 174, 197 Glazunova, Z. I., 250, 263 Gyenes, L., 2, 5, 9, 53, 54

AUTHOR INDEX

H Haase, A. T., 149, 199, 200 Haber, E., 79, 81, 98, 132, 133, 134, 155, 204 Habich, von H., 98 Habichi, G. S., 187, 190, 196 Hall, J. G., 104, 114, 120, 197, 199 Hall, P., 42, 54 Halliday, W. J., 127, 129, 199 Halpern, B., 107, 199 Hamilton, L. D., 130, 171, 184, 194, 199 Han, S., 126, 153, 199, 229, 230, 263 Hanks, J. H., 232, 263 Hannestad, K., 63, 98 Hansson, L. A., 39, 54 Harboe, M., 62, 63, 68, 98, 99 Harding, B., 120, 121, 185, 201 Hardy, D. A., 113,199,206 Harris, J. E., 110, 169, 199 Hart, P. D., 215, 229, 263 Harvey, 3. J., 227, 263 Hasek, M., 120, 195 Haskill, J. S., 187, 199 Hassig, A., 99 Hathaway, W. E., 259, 263 Hayashi, H., 278( 61), 310 Hayat, M., 176, 202 Hayes, C., 142, 145, 202 Haynes, H. A., 229,261 Hayry, P., 117, 164, 173, 199 Hayward, B. J., 4, 51 Hazard, J., 107, 199 Heather, C. J., 170, 206 Hedrick, S. R., 108, 129, 196 Heilman, D. H., 102, 115, 123, 199, 201 Heisch, R. B., 247, 264 Heise, E. R., 126, 139, 143, 153, 157, 199 Hellstrom, I., 113, 116, 121, 170, 199 Hellstrom, K. E., 113, 116, 121, 170, 199 Helmstein, K., 113, 195 Hemmingsen, H., 186, 206 Henney, C. S., 113, 199 Henry, C., 160, 200 Henson, P. hl., 280(72), 285(72), 302 (72), 305( 138), 310, 312 Hepner, G. W., 39, 53 Heremans, J. F., 4, 5, 14, 19, 29, 39, 54

321

Hersh, E. M., 110, 169, 199 Heyneman, D., 247, 263 Hildreth, E. A., 170, 208 Hill, W. C., 132, 160, 171, 199 Hiller, M. C., 272( lo), 299( l o ) , 308 Hilschmann, N., 84, 98 Hirsch, M. S., 227, 263 Hirschhorn, K., 165, 107, 144, 146, 164, 194. 198. 199 Hirschhorn, R., 105, 195, 199 Ho, M., 149, 199, 200 Ho, M-F., 234, 235, 262 Hoare, C. A., 238,263 Hobbs, J. R., 39, 53, 108, 206, 246, 259, 263, 265 Hogarth-Scott, R. S., 42, 43, 49, 53 Hogman, C . F., 31, 37, 39, 40, 53, 54 Holborrow, E. J., 284(86), 311 Holm, G., 112, 113, 115, 117, 118, 119, 120, 121, 138, 141, 175, 200, 203, 204 Holmberg, K., 32, 54 Holmes, E. C., 129, 202 Holt, L., 108, 206, 259, 266 Holtzer, J. D., 152, 154, 200 Hong, R., 39, 40, 51, 52, 108, 196, 259, 262 Hood, L. E., 59, 74,84,98 Hoogstraal, H., 247, 263 Hook, E. W., 149, 200, 207 Hopper, J. E., 63, 66, 67, 68, 69, 71, 75, 76, 83, 84, 85, 90, 93, 97, 98, 99 Hornbrook, hl. M., 2, 3, 4, 16, 53 Housley, J., 20, 22, 23, 55 Howard, A. W., 45,52 Ho\vard, W. L., 259, 263 Howson, W. T., 127, 141, 144, 157, 160, 197 Hoyer, L. W., 275( 2 3 ) , 309 Hnber, H., 126, 153, 200, 205, 299 (117), 312 Huldt, G., 49, 53 Humphrey, J. H., 4, 22, 23, 54, 55, 176, 200, 246, 259, 263, 280( 69), 310 Hunt, W. B., 173, 200 Hunter, A., 45, 52 Hurez, D., 38, 54, 63, 89, 93, 98, 99

322

AUTHOR INDEX

I

K

ldse, O., 272(9), 300(9), 308 linperato, P. J., 248, 263 Irunberry, J., 246, 263 Isaacs, A., 148, 200 Ishizaka, K., 2, 3, 4, 8, 16, 35, 39, 40, 45, 51, 52, 53, 54, 55, 147, 200, 275(25, 29), 280(66), 301(120), 308( 122, 124, 127), 305( 127), 309,

Kaliss, N., 233, 263 Kalpaktosogloa, P., 154, 197 Kaltreider, H. B., 146, 200 Kantor, F. S., 166, 170, 183, 198, 204 Kaplan, A. P., 278(55a), 310 Kaplan, H., 108, 124, 183, 206 Kaplan, J. M., 199 Karush, F., 186, 200 Kasakura, S., 143, 200 Kasel, J. A., 149, 199, 200 Katz, A. B., 132, 133, 134, 155, 204 Katz, D. H., 188, 189, 191, 200, 203 Kau, S. T., 229, 263 Kay, A. B., 281(78a), 284(78a), 311 Kay, W., 165, 171, 200 Keller, R., 302( 123), 312 Kellina, O., 249, 263 Kelly, R., 129, 194 Kelus, A. S., 58, 59, 63, 65, 71, 75, 87. 94, 97, 98 Kempe, C. H., 259, 263 Kennedy, J. C., 190, 191, 193, 200 Kerdel-Vegas, F., 239, 243, 262 Kersey, J., 179, 202 Kesztyus, L., 291( 110), 312 Kettman, J., 188, 200 Keystone, E., 302( 125), 312 Khanolkar, V. R., 210, 213, 216, 263 Khati, B., 246, 263 Khedair, M., 246, 263 Kibrick, S., 105, 150, 199, 204 Killander, D., 105, 200 Killander, J., 31, 32, 39, 53, 54, 65, 91,

310, 312

Ishizaka, T., 2, 3, 4, 8, 16, 39, 45, 52, 53, 147, ZOO, 275( 25, 29), 280( 68), 301( E O ) , 302( 122, 124), 309, 310, 312

Ito, K., 10, 53 Iverson, G. M., 182, 200

J Jamison, D. G., 214, 227, 263, 266 Jancovic, B. D., 245, 263 Jandl, J. H., 154, 201, 276(44), 285 ( 4 4 ) , 289(94), 299(44, 44a, 44b), 306, 309, 310, 311, 312 Janis, M., 145, 194, 200 Jankovic, B. D., 179, 193 Janoff, A., 283(82), 311 Jaton, J. C., 79, 81, 98 Jerne, N. K., 103, 160, 161, 163, 167, 188, 200, 204 Jimenez, L., 133, 163, 166, 167, 168, 169, 185, 194, 195, 200 Johanovsky, J., 123, 127, 129, 131, 135, 136, 157, 194, 201, 203, 206 Johansson, S. A., 278(59), 310 Johansson, S. G. O., 2, 4, 5, 8, 9, 10, 15, 16, 20, 22, 23, 25, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 42, 43, 44, 45, 46, 48, 49, 50, 51, 52, 53, 54, 55, 275( 29), 280( 66), 309, 310 Johnson, J. S., 149, 167, 199, 203 Johnson, R. A,, 280(75), 310 Jonas, W. E., 45, 52 Jonasson, O., 278( 57), 310 Jones, M. A. S., 162, 201 Jopling, W. H., 211, 239, 265 Juhlin, L., 37, 45, 49, 53, 54 Jureziz, R. S., 127, 129, 206

98, 99

Killian, M., 165, 207 Kind, L. S., 167, 200 Klein, E., 113, 172, 174, 175, 200, 204, 206, 207

Klein, G., 139, 172, 200, 201 Klopfenstein, M. D., 259, 263 Klostermann, H., 163, 193 Klun, M. J., 274, 309 Knapp, P. H., 291( 106), 311 Kniker, W. T., 290( 95), 311 Kobayashi, S., 107, 198 Kochwa, S., 40, 54 Koffler, D., 304( l a ) , 312 Kohler, H., 84, 98

AUTHOR INDEX

Kohler, P. F., 35, 54 Kolb, W. P., 137, 139, 141, 175, 198, 201 Kolin, A , , 201 Koller, P. C., 105, 179, 189, 196 Korngold, L., 61, 62, 98 Koss, L. G., 135, 193 Kourilsky, R., 165, 203 Kranier, J., 138, 199 Kraner, K. L., 180, 205 Krause, R. M., 70, 71, 79, 80, 97, 98 Krejci, J., 157, 201, 203, 204 Kronnian, B. S., 129, 201 Kuhns, W. J., 51, 54 Kunkel, H. G., 16, 29, 39, 53, 54, 58, 61, 62, 63, 64, 65, 70, 74, 84, 88, 91, 92, 93, 97, 98, 99, 236, 263, 266, 272, 274( 19), 304( 134), 308, 309, 312

Kunz, M. L., 107, 198 Kuper, S. W. A., 216, 263 Kurban, A. K., 241, 263 Ky, N. T., 107, 199

249,

108, 141, 202,

205, 207

Laymon, C. W., 260,263 Lebowitz, A., 141, 201 Lecks, H. I., 291(105), 311 Leddy, J. P., 272(7), 299(115), 308, 312

Leduc, E. H., 300( 118), 312 Lee, J. C., 115, 193 Lee, L., 307( 145), 313 Lee, S., 171, 197 Lefford, M. J., 177, 194 Lehnian-Fascius, H., 237, 263 Leitoner, A., 272( lo), 299( l o ) , 308 Lelievre, L., 94, 98 Lemma, A,, 239, 241, 244, 248, 263 Lennox, E. S., 190, 191, 193, 200 Leon, M. A., 73, 99, 118, 157, 204, 205 Lepow, I. H., 278(53), 282(53), 283 ( 5 3 ) , 310 Lerner, R. A., 303( 1311, 312 Leskowitz, S., 176, 183, 198, 201 Leuchars, E., 105, 118, 161, 179, 187, 189, 196, 197 Levene, G. M., 260, 264 Levey, D., 275(32), 309 Levin, A. S . , 85, 98 Levine, B. B., 273(14, 15), 300(14, 15), 308

I Lachmann, P., 131, 132, 173, 193 Lackovic, V., 149, 195, 202 Lafferty, K. J., 162, 201 Lage, H. A., 264 Lagrue, G., 107, 199 Lahiri, S. C., 278(56), 310 Lainson, R., 243, 244, 246, 247, 257, 258, 259, 261, 263 Lambert, P. B., 174, 201 Lamvik, J. O., 213, 263 Lamy, L., 245, 261 Lance, E. M., 171, 198 Landsteiner, K., 181, 188, 201 Landy, M., 102, 201 LaPorte, R., 170, 201 Lassus, A., 236, 264 Lawrence, H. S., 102, 103, 106, 124, 125, 126, 127, 129, 130, 144, 167, 191, 193, 196, 201,

323

Levis, W. R., 110, 169, 201 Levy, D. A., 280( 65), 310 Levy, H. B., 149, 199 Lewis, M. R., 102, 122, 204 Liacopoulos-Briot, M., 161, 203 Lichtenstein, L. M., 280(65, 67), 310 Lid&, S., 171, 184, 201 Lieberman, R., 59, 67, 72, 94, 99 Lind, A., 235, 236, 264 Lindeberg, L., 38, 55 Lindenmann, J., 148, 200 Ling, N. R., 104, 105, 113, 199, 201 Linscott, W. D., 299(117), 312 Lipari, R., 61, 98 Litt, M., 284( 89), 311 Livergood, D., 167, 203 Lockshin, M. D., 146, 201 Lo Buglio, A. F., 154, 201, 276(44), 285 ( 4 4 ) , 299(44, 44a), 309 Loeschke, H., 237, 263 Loewi, C., 107, 184, 193, 201 Lolekha, S., 136, 201 Lowenstein, L., 143, 200 Lubaroff, D. M., 171, 201 Lundgren, G., 118, 143, 201, 202 Lundkvist, U., 48, 53 Luparello, T., 291( log), 312 Lycette, €3. R., 106, 203

324

AUTHOR INDEX

Lyon, G., 39, 55 Lyons, H. A,, 291(109), 312

M Mabalay, M. C., 227, 263 McBride, R. A., 104, 162, 181, 191, 201, 205

McCarthy, M. M., 187, 191, 193, 197 McCarthy, J. S., 176, 198 McCloskey, J. W., 171, 201 McClure, D., 107, 208 McCluskey, R. T., 171, 196, 201, 207, 278( 6U),305( 139), 310, 312 McCormick, J. N., 83, 85, 99 MacDonald, A. B., 66, 67, 68, 69, 71, 75, 76, 77, 78, 97, 98 McFadden, E. R., Jr., 291( log), 312 McFarland, W., 110, 114, 115, 201, 205 McGregor, D. D., 104, 105, 155, 190, 196

McGregor, I. A., 42, 54 McGuigan, J. E., 2.84, 311 Macieira-Coelho, A., 142, 143, 151, 198, 199

McIntyre, 0. R., 40, 54 Mackaness, G. B., 135, 177, 194, 201 Mackay, I. R., 161, 197, 237, 265 McKean, D. J., 59, 74, 98 Mackey, G., 28, 29, 54 McLaughlin, C. L., 91, 99 Maclaurin, B. P., 137, 201 McLean, E. P., 129, 195 MacLean, L. D., 110, 143, 198 MacLennan, I. C. M., 120, 121, 185, 201 McRae, D. H., 212,265 Maillard, J. L., 307( 141, 142), 312, 313 Ma'ni, R. N., 144, 201, 247, 262 M..jundar, T. D., 247, 264 Makela, O., 161, 201 Makela, V., 38, 55 Makinodan, T., 76, 98, 160, 162, 189, 201, 203

Malak, J. A., 241, 263 Malchow, H., 114, 119, 178, 293 Mallory, T. B., 170, 197 Malmgren, R. A., 129, 176, 202 Mancini, G., 4, 5, 14, 19, 29, 54 Mandel, T., 161, 202 Mandema, E., 62, 97 Manni, J. A., 121, 204

Mannik, M., 58, 63, 64, 65, 70, 88, 93, 97, 98

Manson, L. A., 164, 205 Manson-Bahr, P. E. C., 237, 244, 246, 247, 248, 249, 264, 265 March, J. T., 291( 107), 312 Marchant, R., 187, 196 Marcus, P. I., 166, 167, 195, 200 Marcus, Z., 123, 126, 202 Margolies, M. N., 79, 81, 98 Margolis, S., 149, 199 Marrone, J., 129, 202 Marsh, B., 111, 144, 193 Marshak, L. C., 260, 264 Marshall, A. H. E., 103, 179, 202 Marshall, W. H., 106, 108, 167, 202 Martin, D., 104, 206 Martinez, C., 179, 202 Martins, A. B., 165, 202 Martins, J. M., 246, 264 Math&, A. A,, 291( 106), 311 MatliC, G., 176, 202 Matisova, E., 202 Matoltsy, M., 135, 207 Matsuoka, Y., 94, 98 Matthew, M., 141, 157, 160, 197 Matthews, L. J., 236, 264 hlatzinger, P., 138, 199 Mauel, J., 113, 115, 119, 120, 173, 195, 202

Mayrink, W., 249, 261 Mazonni, M., 246, 263 Mazzei, D., 231, 264 Meacock, S. C. R., 160, 202 Medawar, P. B., 111, 170, 198, 202, 229, 261

Medina, R., 244, 264 Mehrotra, T. N., 62, 98 Mellbin, T., 42, 53 Melli, G., 231, 264 Melnick, J. L., 151, 163, 167, 197, 207 Merigan, T. C., 149, 150, 202 Merrill, D., 29, 52 Meshaka, G., 63, 89, 93, 98, 99 Metzger, H., 19, 54, 103, 202 Meuwissen, H. J., 108, 179, 180, 196, 202, 259, 262 Meyers, 0. L., 127, 129, 205 Michel, M., 58, 59, 63, 64, 69, 70, 71, 75, 78, 81, 82, 83, 84, 98

325

AUTHOR INDEX

hlichie, D., 167, 196 hliliaesco, C., 63, 89, 93, 98, 99 hliliaesco, E., 38, 54 Miller, E., 127, 129, 206 Miller, F., 19, 54 hliller, H. C., 190, 202, 255, 264 hliller, J. F. A. P., 103, 104, 179, 186, 187, 202, 203 Miller, L. L., 298( 114), 312 Mills, J. A., 106, 202 Milstein, C., 84, 98 Milton, J. D., 231, 264 hlinden, P., 276( 3 7 ) , 309 hlintz, B., 173, 202 hlirsky, A. E., 105, 204 Mishell, R. I., 164, 187, 189. 191, 193, 196, 197, 202 Mitchell, C. F., 186, 187, 196, 202, 203 Mitchison, N. A,, 154, 188, 190, 202 Moberger, G., 113, 195 hloclnbber, F., 162, 202 hlogford, H. E., 260, 261 hloller, E., 115, 120, 121, 143, 145, 161, 189, 190, 197, 199, 202 Moller, G . , 118, 143, 145, 147, 161, 189, 190, 197, 199, 201, 202 hfoller, K. L., 37, 52 Mollison, P. L., 272( I l ) , 274( 2 0 ) , 2885 ( 9 1 ) , 288(91), 297(91), 308, 309, 311 hlontes-Montes, J., 2332, 265 hlontilio, B., 257, 261, 263, 266 hlookerjee, B., 146, 202 Moon, H. D., 112,205 hlooney, J. J., 135, 202 hloore, G. E., 94, 98, 138, 199 hloore, V., 84, 98 Moore, W. D., 165, 202 Moorhead, J. F., 115, 142, 145, 201, 202 hloran, 1%'. C., 280(75), 310 Morgan, H. R., 136, 205 hlorikawa, S., lG2, 202 hforley, J., 141, 157, 160, 197 hlorrell, A., 50, 54 hlorric, B., 104, 203 Morse, H. C., 276(40), 309 Morton, D. L., 113, 116, 129, 176, 202, 205 hfoscarello, hl., 29, 52 Moser, H. W., 39, 55

Mosier, D. E., 168, 187, 202 hloss, R. H., 291( 108), 312 Alota, I., 275(31), 305( 136), 309, 312 hlouton, D., 161, 189, 194 hlovat, 11. Z., 287(93), 302( 130), 311, 312 Slonbray, J. F., 231, 264 hloyne, hf. A., 94, 98 hludd, S., 127, 206 hlueller, G. C., 105, 119, 198 Xfukherjee, A. C., 246, 264 hlukherjce, A. hl., 247, 265 hluller, J. Y., 110, 189, 194 hfiiller-Eberhard, H. J., 121, 204, 277 ( 4 7 ) , 278(47, 5 5 ), 282(55), 299 ( 1 1 7 ) , 310, 312 hlunoz, J., 290( 102), 311 Sluschel, L. H., 274, 309 Alustard, J. F., 302( 130), 312 hlustakallio, K. K., 236, 264 Slyers, hl. LV., 129, 202 hlynors, L. S., 45, 52 hlyrvik, Q. W., 127, 129, 173, 198, 200

N Nachnlan, R. L., 62, 91, 99 Naff, C.. B., 279( G2),310 Nahmias, A. J., 259, 264 Najarian, J. S., 171, 197, 203 Saor, D., 161, 203 "ase, S., 188, 204 Naspitz, C. K., 104, 105, 203 Navalkar, R. G., 235, 236, 264 Navarro, E., 216, 265 Neal, R. A., 246, 249, 264 Nelken, D., 244, 261 Nelson, C. T., 278(58), 310 Selson, D. S., 134, 152, 155, 171, 203, 231, 264, 276(42), 278(51), 285 (42, 51), 309, 310 Nelson, hl., 231, 264 Nelson, R. A., Jr., 278(52), 310 Seogy, K. N., 246,264 Ncry-Guimnrez, F., 249, 264 Neveu, T., 245, 264 Newell, K. W., 227, 233, 264 Niall, H. D., 29, 52 Nilsson, H., 118, 204 Nilsson, K., 42, 54 Nishio, J., 126, 129, 136, 205

326

AUTHOR INDEX

Nisonofl, A., 66, G7, 68, 69, 71, 75, 76, 77, 78, 85, 86, 88, 90, 93, 97, 98, 99 Nissen, B., 132, 199 Noguchi, H., 260, 264 Norberg, R., 50, 51, 54 Nordbring, F., 40, 54 Nordin, A. A,, 114, 115, 116, 117, 121, 160, 173, 179, 180, 195, 196, 200 Nordqvist, B., 129, 136, 203 Norlin, M., 235, 236, 264 Notth, R. J., 171, 184, 203 Nossal, G . J. V., 82, 98, 161, 187, 201 203 Nota, N. R., 161, 203 Notani, G., 72, 94, 97 Novnles, J., 216, 265 Nowell, P. C., 104, 162, 163, 203, 207 Nussenzweig, V., 110, 194, 299( 116), 312

0 Oakley, C. L., 237, 261 Oettgen, H. F., 129, 167, 168, 195, 200 Ogawa, hl., 40, 54 Ogston, D., 279( 6 2 ) , 310 bhnian, S., 37, 54 Ohms, J., 84, 99 Old, L. J., 129, 153, 173, 176, 194, 195, 203 Olds, R. S. J., 123, 153, 193 Oldstone, hl. B., 138, 203 Olhngen, B., 50, 51 Oliveirn, B., 274( 18), 300( 18), 309 Oort, J., 104, 203, 218, 266 Oppcnheim, J. J., 106, 110, 203, 205 Orange, R. P., 275( 34), 276( 38, 40a), 282( 40a), 284( 8 8 ) , 287(88), 305 ( 3 4 ) , 309, 311 0 1 tolani, C., 231, 264 Osler, A. G., 274( 1 8 ) , 275( 32), 280( 65, 73), 300( 18), 302(73), 309, 310 Osobn, D., 103, 104, 179, 189, 198, 202, 203 Osserman, E. F., 274( 1 9 ) , 309 Oochterlony, O., 235, 236, 264 Oudin, J., 58, 59, 63, 64, 69, 70, 71, 75, 78, 81, 82, 83, 84, 97, 98 Ovary, Z., 107, 188, 203, 271(5), 275 ( 3 6 ) , 280(36), 308, 309

Oviedo, B. V. E., 248, 265

P PackalBn, T., 113, 120, 147, 179, 204, 207 Palmer, C. G., 167, 203 Palmer, E., 214, 215, 226, 263, 265, 266 Panayi, G. S., 141, 157, 160, 197 Pang, 11. K., 137, 204 Papermaster, B. W., 179, 202 Pappenheinier, A. M., Jr., 123, 186, 203, 206 Parnense, L. W., 250, 264 Paraf, A., 94, 98 Parnskova-Tchernozet~ska, E., 142, 145, 202 Parish, W. E., 304(132a), 312 Parrott, D. M. V., 260, 264 Parrow, A., 39, 52 Passaleva, A., 123, 129, 204 Patrick, R. A., 110, 194, 278(53), 282 ( 5 3 ) , 283( 53), 310 Patterson, R. J., 136, 177, 203 Paul, C., 84, 98 Paul, W. E., 188, 189, 191, 200, 203 Penrce, J. D., 107, 197 Pearlman, D. S., 259, 263, 280, 310 Peartiinin, G., 106, 203 Pearson, J. M., 226, 231, 264, 265 Pearson, hl. N., 176, 203 Peary, D., 111, 144, 193 Pederson, N. C., 104, 203 Pekarek, J., 127, 129, 131, 135, 136, 157, 194, 201, 203, 206 Pelley, R. P., 157, 205 Peltier, A. P., 165, 203 Penn, G. M., 84, 99 Pensky, J., 279(62), 310 Pepys, J., 44, 54, 218, 224, 235, 236, 265 Perey, D. Y. E., 154, 180, 197, 203 Perhnm, R. N.,59, 73, 97 Perkins, E. H., 162, 203 Periman, P. O., 166, 196 Perlniann, H., 113, 120, 121, 204 Perlmnnn, P., 37, 53, 112, 113, 115, 117, 118, 119, 120, 121, 138, 139, 141, 175, 195, 200, 201, 203, 204 Pernis, B., 165, 204 Peterson, R. D. A,, 102, 103, 179, 180, 193, 196, 204

327

AUTHOR INDEX

Peterson, W. A., 157, 200 Pettit, J. H. S., 241, 264 Pfeiffer, E. F., 165, 197 Phair, J. P., 183, 204 Phillips, hl., 276, 310 Phillips, S. hl., 170, 208 Pick, E., 157, 204 Pierpaoh, W., 290(98), 311 Pierce, G. E., 113, 116, 121, 199 Pilchard, E. I., 129, 204 Pinardi, h l . E., 243, 248, 249, 262 Pincus, W. B., 137, 138, 139, 143, 157, 204 Pink, J. R. L., 84, 99 Piper, P. J., 281( 78), 310 Pirrie, A., 142, 145, 202 Pitombeira, M. da S., 246, 264 Plantin, L. O., 50, 51 Playfair, J. H. L., 186, 205 Plotz, P. H., 275(24), 309 Pochyly, D. F., 130, 204 Pogo, B. G . T., 105, 204 Polak, L., 234, 264 Polgar, P. R., 105, 204 Polley, M. J., 299( 117), 312 Pollock, T. M., 260, 261 PontCn, J., 42, 54 Porath, J., 10, 29, 51, 54, 55 Porter, R. R., 20, 54 Potter, M . , 59, G7, 72, 73, 74, 94, 98, 99 Prendergast, R. A., 134, 171, 180, 204, 205 Pressman, D., 76, 94, 98, 99 Preston, P. M., 239, 241, 244, 248, 251, 252, 254, 2.57, 259, 262, 263, 264 Preudhomnie, J.-L., 38, 54 Price, E. W., 239, 242, 264 Priolisi, A., 246, 264 Privat, Y., 260, 265 Prouvost-Danon, A., 275( 35, 35a), 280 ( 6 8 ) , 302(68), 309, 310 229, 264 Ptak, W., Putnam, F., 84, 98 Puttrell, C. N., 149, 207

Q Quie, P. G., 108, 196, 2.59, 262 Quilici, M., 246, 248, 249, 259, 260, 264, 265

R Radovich, J., 186, 206 Radwanski, Z., 248, 255, 256, 257, 258, 262, 264 Radzimski, G., 76, 99 Raff, M. C., 114, 179, 204 Raffel, S., 126, 129, 149, 165, 176, 198, 202, 203, 204 Rahim, G. A. F., 239, 244, 261, 264 Raidt, D. J., 187, 191, 193, 197 Rajapakse, D. A., 129, 204 Rajewsky, K . V., 188, 204 Ramseier, H., 157, 159, 204 Ranadive, N. S., 283(85), 311 Rahim, G. A. F., 239, 244, 261, 264 265 Rapoport, S. I., 298(112), 312 Rapp, H. J., 129, 175, 176, 201, 207, 275(23, 25), 280(69), 309, 310 Rasanen, ." J. A., 38, 55 Rassmussen, A. F., Jr., 290( loo), 291 (107), 311, 312 Ratnoff, 0. D., 279(50, 62), 310 Rauch, H . C., 165, 204 Rawls, W. E., 127, 129, 151, 163, 167, 177, 197, 206, 207 Rebonato, C., 248, 249, 262 Reed, C. E., 291( 104), 311 Reerink-Broncers, E. E., 43, 54 Rees, R. J. W.,212, 213, 214, 215, 218, 224, 225, 226, 227, 229, 232, 235, 236, 242, 262, 263, 264, 265, 266 Reid, R. T., 276(37), 309 Reif, A. E., 114, 179, 204 Reiqnam, C. W., 259, 263 Reisfeld, R. A., 133, 204, 207 Reizenstein, P., 50, 51 Reisman, R. E., 273(16), 300(16), 301 ( E l ) , 309, 312 Remold, H . G., 132, 133, 134, 148, 155, 204, 207 Renolcl, A. E., 165, 197 Report of the Panel on Bacteriology and Immunology, 215, 265 Revesz, L., 172, 175, 200, 204 Rezai, H . R., 257,258,265 Ricci, M., 123, 129, 204 Rice, S. A., 72, 94, 97 Rich, A. R., 102, 122, 204 '

328

AUTHOR INDEX

Riches, H. R. C., 108, 206, 259, 266 Richter, M., 104, 105, 186, 203, 204 Ridley, D. S., 211, 212, 222, 223, 233, 239, 242, 265, 266 Rieke, W. O., 165, 171, 200 Rigler, R., 105, 200 Ringertz, N. W., 175, 200, 204 Ripps, C.,107, 198 Robbins, J. H., 110, 169, 201 Roberts, K. B., 106, 202 Robertson, C. L., 72, 99 Robinson, J. O., 275(30), 309 Rockey, J. H., 16, 39, 54 Rocklin, R. E., 108, 127, 129, 141, 185, 204, 205 Rodriguez, O., 216, 265 Roholt, 0.A., 76, 99 Rosenau, W., 112, 113, 115, 116, 193, 199, 205 Romagnani, S., 123, 129, 204 Romero, J., 244, 264 Rorsman, H., 129, 136, 203 Rosak, M., 185, 206 Rose, B., 290( 99), 311 Rose, N. R., 107, 198, 273(16), 300 (16), 309 Roseman, J. M., 189, 205 Rosen, F. S., 299(44b), 310 Rosenau, W., 112, 113, 115, 116, 193 205 Rosenberg, E. B., 43, 54, 55 Rosse, W. F., 274, 309 Rother, K., 305( 139), 312 Rother, U., 305( 139), 312 Rothman, S., 260, 264 Rouze, P., 94, 98 Rowe, D. S., 2, 28, 29, 31, 32, 39, 42, 52, 54 Rowley, M. J., 237, 265 Rowsell, H. C., 302( 130), 312 Ruddle, N. H., 118, 119, 137, 142, 175, 205 Rudolf, B., 115, 202 Rudolf, H., 113, 115, 119, 120, 173, 195 Rugarli, C., 231, 264 Russell, P. S., 170, 207 Ryder, G., 255, 258, 262 Rytel, M. W., 127, 129, 197

s Sadun, E. H., 248, 262 Sagher, F., 237, 239, 242, 262 Saitz, E. W., 215, 235, 265 Salazar-Mallen, M., 232, 265 Salmon, S. E., 28, 29, 54 Salsbury, A. J., 176, 196, 259, 264 Salvin, S. B., 123, 126, 129, 136, 182, 184, 185, 186, 188, 189, 190, 205, 206 Samloff, I. M., 298( 114), 312 Samuels, H. H., 104, 206 Sandberg, A. L., 274( 18), 300( 18), 309 Sanderson, A. R., 139,205 Sato, T., 162, 203 Saul, A,, 216, 265 Schaller, K. F., 238, 239, 261 ScharE, M. D., 138, 184, 186, 189, 194, 203, 206 Schechter, G. P., 110, 114, 201, 205 Scherrer, R., 94, 98 Schick, B., 269, 272, 308 Schierman, L. W . , 181, 191, 201, 205 Schild, H . O., 159, 207 Schimpl, A,, 187, 205 Schindler, R., 115, 195 Schirrmacher, V., 188, 204 Schlossman, S. F., 124, 129, 132, 183, 196, 205 Schumberger, J. R., 176, 202 Schneider, M., 176, 202 Schoenbechler, M. J., 285( 92), 311 Schoenwetter, W. F., 170, 208 Schooley, J. C., 76, 99 Schrek, R., 106, 205 Schubert, D., 91, 99 Schur, P. H., 304( 134), 312 Schwartz, H. J., 157, 205 Schwartz, R. S., 228,265 Schwartzenberg, L., 176, 202 Seeger, R. D., 110, 205 Segal, H. L., 298( 114), 312 Sehon, A. H., 2, 4, 5, 9, 53, 54, 107, 198, 205

Sela, M., 183, 195 Seligmann, M., 38, 54, 63, 89, 93, 98, 99 Sell, S., 154, 205 Sen Gupta, P. C., 246, 247, 264, 265 SBriB, C., 238, 239, 242, 261, 262

AUTHOR INDEX

Shacks, S. J., 137, 141, 175, 198 Shaninia, A. H., 236, 265 Shaw, J. J., 251, 265 Shea, J. D., 136, 205 Sheagren, J. N., 228, 230, 234, 235, 243, 265, 266 Sheffer, A., 108, 185, 204, 205 Shepard, C. C., 212, 215, 225, 226, 227, 230, 2.35, 262, 265 Sher, R., 258, 265 Shimizu, A., 84, 98 Shin, H. S., 2~74(18), 300( 18), 309 Shulnian, N. R., 272( lo), 277(46), 299 (10, 46), 308, 310 Shupe, S., 138, 199 Sidorova, E. V., 115, 195 Silvers, W. K.. 162. 173, 194, 202, 207 Silverstein, A. M., 180, 182, 186, 198, 205 Simmons, T., 164, 205 Simon, N. J., 236, 263 Simonsen, hl., 162, 205 Simpson, P. J., 167, 203 S Iskind, '. ' G. W., 188, 203 Sjoberg, H. E., 278(59), 310 Slater, R. J., 58, 61, 99 Slavina, E. G., 150, 196 Smith, C., 40, 54 Smith, R. F., 182, 184, 185, 186, 188, 189, 190, 205 Smith, R. T., 111, 142, 144, 145, 193, 205 Smith, S. J., 42, 54 Smith, T. J., 149, 150, 205 Smyly, H. J., 210, 262 Smyth, A. C., 136, 205 Snyder, R. hl., 149, 207 Sober, H. A., 124, 183, 205 S@borg, hl., 146, 147, 194, 205 Sodrt., A,, 243, 248, 249, 261 Soghor, D., 146, 200 Solheim, B. G., 63, 68,98, 99 Solliday, S., 145, 164, 194, 205 Solomon, A., 91, 99 Solomon, G. F., 2 9 l ( 108), 312 Sonozaki, H., 171, 205 Soothill, J, F., 33, 44, 52 Sorkin, E., 151, 152, 153, 194, 195, 276 ( 4 3 ) , 290( 98), 309, 311 South, M.A,, 179, 196

329

Southgate, B. A,, 248, 249, 264, 265 Spector, W.G., 159, 170, 195, 207 Spencer, E. S., 291(107), 312 Spiegelberg, H. L., 50, 54 Spink, W. W., 307( 144), 313 Spiro, D., 170, 171, 207 Spitler, L. E., 108, 124, 126, 144, 183, 205, 206 Spitznagel, J. K., 198 Spring, S. B., 78, 99 Stainislawski, M., 94, 98 Stanworth, D. R., 2, 4, 8, 20, 22, 23, 51, 52, 54, 55 Stastny, P., 126, 137, 143, 206 Stauber, L., 237, 258, 259, 265 Stechschulte, D. J., 276( 38, 39), 281 (78a), 284(78a), 309, 311 Steininger, W.J., 259, 263 Steel, T., 113, 206 Steffen, C., 165, 206 Stein, S., 62, 91, 99 Steinberg, A. G., 37, 51 Stenius, B., 48, 49, 55 Stern, K., 186, 196, 206 Stetson, C. A., Jr., 307( 145), 313 Stewart, L. C., 207 Stiehm, E. R., 29, 55 Stiffel, C., 161, 189, 194, 203 Stinclxing, \V. R., 149, 207 Stockert, B., 176, 203 Stone, hi. hl., 215, 261 Stone, S. II., 233, 261 Strejan, C., 300( 119), 312 Strickland, G. T., 43, 55 Strober, S., 104, 206 Stupp, Y., 183, 195 Stutnian, O., 179, 180, 202 Sulitzeanu, D., 147, 161, 203, 206 Sundvno, J. S., 138, 199 Sutherland, I., 213, 266 Suzuki, hl., 134, 204 Svejcar, J., 123, 127, 129, 131, 136, 157, 201, 203, 206 Svet-Moldovsky, G. J., 150, 196 Swanson, M. A., 228, 265 Swedluncl, H. A., 29, 53 Sn.ett, V. C., 110, I96 Szenlierg, A., 82, 98, 207 Szentivanyi, A,, 290( 101), 291, 311

330

AUTHOR INDEX

T Tada, T., 35, 55 Tai, C., 284, 311 Taichmnn, N. S., 287(93), 302( 130), 311, 312

Takahashi, T., 187, 206 Takasugi, M., 113, 206 Takiguchi, T., 111, 144, 193 Talal, N., 275( 24), 309 Taliaferro, W. H., 76, 99 Talniage, D. W., 76, 99, 186, 206 Talwar, G. P., 230, 265 Tan, E. M., 272, 308 Tan, hi., 91, 99 Tanaka, N., 300( 118), 312 Tannahill, A. J., 260, 261 Tartar, I. H., 239, 264 Tauaka, T., 176,207 Taubler, J. H., 127, 206 Taylor, J. B., 146, 200 Taylor, R. B., 186, 187, 188, 206 Tee, R. R., 218, 224, 235, 236, 265 Temine, P., 260, 265 Temple, A., 107, 201 Ternynck, T., 10, 12, 51 Terry, W. D., 2, 50, 52, 54, 274( 19, 2 0 ) , 276(41), 283(111), 309, 312 Tliieffry, S., 39, 55 Thomas, L., 102, 124, 125, 126, 127, 129, 130, 193, 196, 206 Thomas, M.-T., 142, 151, 199 Thompson, E. B., 104, 206 Thor, D. E., 108, 127, 129, 196, 206 Thorbecke, G. J., 187, 206 Thurston, J. M., 231, 264 Thyresson, N., 37, 54 Tkachyk, S. J., 29, 52 Toft, B., 29, 52 Tomasi, T. B., Jr., 301( 121), 312 Tomioka, H., 147, 200 Tomlinson, A. S., 306( 140), 312 Tompkins, G. M., 104, 206 Tompkins, W. A. F., 104, 127, 129, 177, 206

Torrealba, J. F., 243, 262 Torrealba, J. W., 243, 247, 262, 265 Torrigiani, G., 119, 199, 186, 205 Touillet, F., 160, 207 Townes, A. S., 235, 265 Tramier, G., 260, 265

Trautman, J. R., 228, 230, 234, 235, 236, 243, 264, 265, 266 Treadwell, P. E., 190, 191, 193, ZOO, 284( go), 290( l o o ) , 306( go), 311 Tregear, G. W., 29, 52 Trernonti, L., 247, 266 Triplett, R. F., 187, 196 Trujillo, 0. D., 258, 266 Tseng, J. J., 239, 263 Tsoi, hl., 173, 175, 206 Turcotte, R., 107, 198 Turk, J. L., 104, 111, 136, 152, 157, 165, 170, 171, 176, 184, 194, 196, 197, ZOO, 203, 204, 206, 216, 217, 218, 219, 220, 221, 223, 224, 228, 229, 231, 232, 234, 235, 242, 243, 251, 252, 253, 255, 260, 262, 263, 264, 266

Turner, Turner, Turner, Tnrohy,

K. J., 105, 206 M. D., 298( 114), 312 M. W., 32, 34, 42, 55 D. \\’., 255, 264, 266

U Uhr, J. W., 104, 107, 123, 144, 146, 184, 186, 189, 194, 198, 203, 206, 276, 310

Ulrich, M., 258, 266 Unanue, E. R., 176, 206, 272(13), 273 ( 1 3 ) , 274( 13), 288( 13), 303(13), 308

Ungar, G., 278(61), 309 Urihara, T., 302( 130), 312

V Vahlqvist, B., 42, 53 Valdiniarsson, H., 108, 206, 259, 266 Valentine, F. T., 106, 108, 144, 167, 202, 207

Valentine, M. D., 280(74), 284( 8 8 ) , 287( 881, 302( 74), 310, 311 Valentine, R. C., 67, 99, 212, 265 Van Arsdel, P. P., Jr., 107, 208 Vane, J. R., 281(78), 310 Van Furth, R., 171, 207 Van Leeuwen, G., 61, 62, 98 Vannier, W. E., 16, 51, 275( 23), 309 Vassali, P., 171, 207, 278(60), 305 (139), 310, 312 Vaughn, J. H., 123, 129, 198, 272, 308

AUTHOR INDEX

Vaz, N. M., 275(36), BO(36, 68), 302 (68), 309, 310 Veach, S. R., 127, 129, 206 Venancio, I. A., 264 Vischer, T. L., 107, 201 Vogt, W., 282(81), 283(82), 311 Voisin, G. A., 160, 207, 307( 141, 142), 312, 313

Voisin, J., 160, 207 Volknian, A., 155, 171, 196, 207 Voller, A,, 42, 43, 49, 53, 55, 251, 265 Volluni, R. L., 227, 263 van Kretschmar, W., 250, 258, 266 von Piquet, C., 268, 269, 308 Voynow, hl. K., 146, 184, 193

W Wade, H. W., 210, 214, 266 Wager, O., 38, 55, 236, 264, 266 Wagner, R. R., 149, 150, 200, 205, 207 Wahren, B., 40, 54 Waksnian, B. H., 118, 119, 135, 137, 142, 170, 171, 175, 179, 193, 201, 202, 205, 207

Waldman, T., 50, 54 Waldmann, T. A., 50, 55 Waldorf, D. S., 228, 243, 266 Wallace, F. G., 238, 263 Wallin, J., 113, 199 Wallis, V. J., 105, 118, 161, 179, 187, 189, 196, 197 Walton, B. C., 247, 249, 266 Waltraut, H. L., 299(116), 312 Wang, A. C., 63, 83, 84, 85, 90, 93, 99 Wang, I. Y. F., 83, 85, 99 Ward, P. A., 148, 207, 278(54), 280(71, 7 7 ) , 282(54), 284(87c), 310, 311 Ward, S. M., 58, 61, 99 Warner, N. L., 207 Warren, A. K., 167, 203 Wasserman, J., 113, 120, 147, 179, 204, 207

Waterfield, M. D., 79, 81, 98 Waters, M. F. R., 211, 212, 213, 215, 216, 217, 220, 221, 223, 224, 225, 228, 229, 231, 233, 234, 235, 242, 243, 262, 264, 265, 266 Watten, R. H., 43, 55 Webb, M., 127, 129, 199 Webster, M., 277( 48), 282(48), 310

331

Wecker, E., 187, 205 Weddell, A. C. M., 214, 225, 226, 263, 265, 266

Weigert, hl. C., 59, 73, 99 Weigle, W. O., 76, 99, 187, 190, 196, 305( 137), 312 Weil, hf. H., 307( 144), 313 Weiser, R. S., 126, 139, 143, 153, 157, 173, 175, 198, 199, 206, 229, 263 Weiss, L., 136, 205 Weissman, C., 105, 195, 199 Weksler, M., 231, 264 Welzer, A,, 278( 58), 310 \Vemambu, S. N. C., 224, 234, 235, 242, 266

Wepsic, H. T., 129, 175, 201, 207 Werner, B., 117, 200 Wertheim, G., 257, 266 \Vessl&i, T., 104, 156, 207 \Vhalen, C. E., 43, 54, 55 Wheelock, E. F., 149, 166, 197, 207 White, J. G., 138, 199 \Vhite, R. G., 105, 179, 202 Wicher, K., 10, 53, 301(121), 312 Wide, L., 29, 39, 45, 48, 49, 53, 54, 55 Wiener, J., 170, 171, 207 Wigzell, H., 190, 207 Wilcox, R. R., 272(9), 300(9), 308 Willenis, F. T. C., 163, 167, 207 Williams, A. C., 174, 207 Williams, K., 42, 54 Williams, R. C., 58, 62, 63, 64, 65, 70, 98, 99

Williams, R. C., Jr., 236, 266 Williams, R. M., 179, 207 Williams, T. W., 137, 138, 140, 141, 175, 198, 207

Willms-Kretschmer, K., 176, 198 Willoughby, D. A., 159, 160, 176, 196, 202, 207

\Vilson, D. B., 112, 115, 162, 163, 207 Wilson, S. K., 63, 83, 84, 85, 90, 93, 98, 99

Winkler, K. C., 152, 154, 200 Wiskott, A., 37, 55 Witebsky, E., 273( 16), 300( 16), 309 Witten, T. A., 165, 207 \Volff, S. M., 230, 234, 235, 265 Wollheim, F. A., 40, 55

332

AUTHOR INDEX

Wolstencroft, R. A., 106, 127, 141, 157, 160, 197, 201, 203, 207, 250, 251, 254, 255, 256, 257, 262 Wong, W. L., 165,207 Wood, D. W., 291( 105), 311 Wood, W. B., Jr., 269(4), 308 Woods, W. W., 137,204 Woodruff, C. E., 259, 263 Wright, D. J. M., 260, 264 Wuepper, K. D., 278( 55b), 310 Wuhrmann, F. H., 60,99 Wunderlich, J. R., 117, 164, 173, 207 Wunderly, C. H., 99

Y Yagi, Y., 94, 98 Yalow, R. S., 298(113), 312 Yaron, A., 124, 183, 205 Yee, C. L., 129, 202 Yonkovich, S. J., 59, 73, 99 Yoshida, K., 259, 264 Yoshida, T., 133, 171, 204, 207

144, 242, 258,

Youmans, G., 13'6, 177, 203 Young, J. D., 108, 124, 183, 206 Young, R. R., 39,55 Youngner, J. S., 149, 207 Yphantis, D. A., 8, 16, 19, 55

Z

195,

Zabriskie, J. B., 147, 207 Zak, S. J., 183, 201 Zarling, J. M., 177, 206 Zbar, B., 129, 175, 176, 201, 207 Zeiss, I., 114, 119, 178, 193 Zeitz, S. J., 107, 208 Zemen, N., 282(82), 283(82), 311 Ziff, M., 126, 137, 143, 206 Zitrin, C. M., 134, 155, 193 Zolov, D. M., 273( 15), 300( 15), 308 Zoschke, D. C., 106, 145, 169, 194, 208 Zuckerman, A., 239, 262 Zukosk, C. F., 143, 201 Zunker, H. O., 171,207 Zvaifler, N. J., 275(30), 309 Zweiman, B., 170, 208

SUBJECT INDEX A

minants

Adjuvants effects, cell-mediated immune reactions and, 176 Allergic reactions, see also Hypersensitivity, and Immediate-type leprosy and, 222-225 Allografts rejection, cell-mediated itiiniune reactions and, 172-175 Amino acid ( s ) sequence antibody persistance and change during prolonged immunization,

Antigen, immediate-type allergic reactions and, 271-273 Antigen-antiglobulin reaction, red celllinked, 4 5 1 6 Antiidiotypic sera cross-reactions different donors, 69-70 identical immunoglobulin molecules in different individuals, 73-75 myeloma proteins, 72-73 same individuals, 70-72

6

79-81 identical immunoglobulins in different individuals, 73-75 Antibody anti-ind, cross-reactions with nonspecific immunoglobulins, 92-93 anti-Salmonello, shared determinants, 81-83 cell-mediated immune reactions and,

151-156 combining site, individually specific antigenic determinants and, 86-87 formation cells involved, 185-193 enumeration of specifically sensitized cells and, 160-162 humoral, production in leprosy, 234-

from same individual,

70-72

Blastogenic

factors, lymphocytes

and,

143-147

C Cell-mediated immune reactions, 102-105 antibody formation, 178 cells involved, 185-193 effector cells in delayed-type hypersensitivity, 179-185 depression in leprosy, 227-234 direct cytotosicity of target cells by lymphocytes, 111-112, 121-122 correlation of, 119-121 niernbrane-associated antigens, 113-

237 idiotypic cross-reaction in different donors, 69-70 immediate-type allergic reactions and,

273-277

117 soluble antigens, 117-119 enumeration of specifically sensitized cells, antibody formation, 160-162 delayed-type hypersensitivity, 165-

169

normal populations, idiotypic specificity for limited heterogeneity, 75 persistence and change during prolonged immunization amino acid sequence evidence, 79-81 idiotypic specificities and, 7G-79 populations, individual antigenic specificities in, 6.3-69 idiotypic deterunrelatedness of

333

mixed Iyniphocyte interaction, 162-

164 lymphocyte transformation, 105-110 cell cooperation and lymphocyte activation, 110-111 mediators antibodies, 151-156 blastogenic factors, 143-147 chemotactic factor, 147-148

334

SUBJECT INDEX

interferon, 148-151 lymphotoxin, 137-143 migration inhibitory factor, 122-137 skin-reactive factor, 156-160 relationships between in vitro and in vivo results adjuvant effects, 176 cellular resistance, 177 histology, 170-172 models, 169-170 rejection of allografts and tumors, 172-175 unified model, 177-178 Chemotaxis, cell-mediated immune reactions and, 147-148 Complement, leprosy and, 234-237

E Enzymes, immediate-type actions and, 277-280

allergic re-

G Guinea pig, experimental leishmaniasis, in, 250-259

H Hypersensitivity delayed-type effector cells, 179-185 enumeration of specifically sensitized cells and, 165-169 histology and, 170-172

I Immediate-type allergic reactions, classification, basis and general description, 291-298 components antibodies, 273-277 antigens, 271-273 cells, 28S285 effector enzyme systems, 277-280 mediators, 281-283 direct responses, 298-299 general, 267-270 indirect responses anaphylactic-type, 299-302 macromolecular mediator-determined, 302-305 unknown mediator-determined, 305

mixing of categories in natural reactions, 305-307 pseudoallergic reactions and, 307-308 sensitization and, 270-271 sites of antigen-antibody reaction, 285286 the terrain, 289-291 time course, 286-289 Immunoglobulin( s ) amino acid sequence, identical molecules in different individuals, 7375 leprosy and, 234-237 nonspecific, cross-reaction with antiincl antibodies, 94 Immunoglobulin E chemical and phy.sica1 characteristics, 15-19 detection of antibody activity allergen antibodies in serum, 48-49 radioallergosorbent test, 46-48 red-cell-linked antigen-antiglobulin reaction, 4 5 4 6 general, 1-2 identification of myeloma protein ND,

2-4 isolation of, 4-15 levels in disease factors influencing, 44-45 secretions, 43-44 serum, 35-43 levels in health secretions, 32-35 serum, 29-32 metabolism, 49-51 methods for determination, 28-29 properties of active regions e chain, 19-20 enzymatic fragments, 20-28 Immunoglobulin( s ) G and M shared idiopathic determinants anti-Salmonella antibodies, 81-83 structural relationships of clonally produced, 83-85 Individual antigenic specificity antibody populations qualitative analysis, 63-66 quantitation of idiotypic specificities, 66-69 general, 58-60

335

SUBJECT INDEX

localization of determinants, 85-86 combining site and, 86-87 isolated heavy and light chains, 87-

91 light

chain variable

region and,

91-92 monoclonal origin, 94 monotypic proteins and, 60-63 Infections chronic, host-determined spectrum of clinical manifestations, 209-210,

259-260 Interferon, cellmediated actions and, 148-151

iminune

re-

L Leishmania enriettii, guinea pig infection,

250-259 Leishmaniasis artificial immunization, 249-250 disease spectrum, 238-247 experimental in guinea pig, 250-259 general, 237-238 imniunodiagnosis, 248-249 leishmanin test, 247-248 Leprosy allergic reactions in, 222-225 clinical spectrum, 210-212 experimental production in animals,

225-227 histological features, 212-214 lymphoid tissue, 216-222 human depression of cellmediated immunity and, 227-234 immunoglobulins, complement and humoral antibody production,

234-237 lepromin test, 214-21 6 Lymphocytes blastogenic factors and, 143-147 direct cytotoxicity of target cells correlation with cell-mediated immunity, 119-121 membrane-associated antigens, 113-

117 soluble antigens, 117-119

mixed interaction, enumeration of specifically sensitized cells and,

162, 164 transformation, cell-mediated immune reactions and, 105-1 11 Lymphoid tissue, histological changes in leprosy, 216-222 Lymphotosin, growth inhibitory factors and, 137-143

M Macrophages, factors affecting, 122-137 Models cell-mediated immune reactions, 169-

170 unified, 177-178 Myeloma proteins, cross-reactivity in different individuals, 72-73 Myeloma protein ND, identification of,

2 4

P Protein( s ) monotypic, individual antigenic specificity and, 60-63 Pseudoallergic reactions, immediate-type allergic reactions and, 307-308

R Radioallergosorbent test, immunoglobulin E class and, 4 6 4 8

S Serum allergen antibodies in, 4-9 immunoglobulin E levels disease, 35-43 health, 29-32 Skin-reactivity, cell-mediated reactions and, 15&160

immune

T Tumors, cell-mediated immune reactions and, 172-175

This Page Intentionally Left Blank