Table Of Contentii
PROCEEDINGS OF THE FIRST INTERNATIONAL PHARMACOLOGICAL MEETING,
STOCKHOLM, 22-25 AUGUST, 1961
Vol. 1 Part 1: Plenary Session
Part 2: Pharmacological Control of Release of Hormones Including Antidiabetic
Drugs
Vol. 2 Effects of Drugs on Synthesis and Mobilization of Lipids
Vol. 3 New Aspects of Cardiac Glycosides
Vol. 4 Drugs and Membranes
Vol. 5 Methods for the Study of Pharmacological Effects at Cellular and Subcellular
Levels
Vol. 6 Metabolic Factors Controlling Duration of Drug Action
Vol. 7 Modern Concepts in the Relationship between Structure and Pharmacological
Activity
Vol. 8 Pharmacological Analysis of Central Nervous Action
Vol. 9 Part 1: Bradykinin and Vaso-dilating Polypeptides
Part 2: Pharmacology of the Lung
Vol. 10 Abstracts
PROCEEDINGS OF THE SECOND INTERNATIONAL PHARMACOLOGICAL MEETING
PRAGUE, 20-23 AUGUST, 1963
Vol. 1 Pharmacology of Conditioning, Learning and Retention
Vol. 2 Biochemical and Neurophysiological Correlation of Centrally Acting Drugs
Vol. 3 Pharmacology of Cholinergic and Adrenergic Transmission
Vol. 4 Drugs and Enzymes
Vol. 5 Pharmacology of Cardiac Function
Vol. 6 Pharmacology of Smooth Muscle
Vol. 7 Pharmacology of Oriental Plants
Vol. 8 Evaluation of New Drugs in Man
Vol. 9 Recent Advances in the Pharmacology of Toxins
Vol. 10 Oxytocin, Vasopressin and their Structural Analogues
Vol. 11 Drugs and Respiration
PROCEEDINGS OF THE THIRD INTERNATIONAL PHARMACOLOGICAL MEETING
SAO PAULO, 24-30 JULY, 1966
Vol. 1 Mode of Action of Anti-Parasitic Drugs
Vol. 2 Pharmacology of Reproduction
Vol. 3 Clinical Pharmacology
Vol. 4 Mechanisms of Drug Toxicity
Vol. 5 77?^ Control of Growth Processes by Chemical Agents
Vol. 6 Drugs in Relation to Blood Coagulation, Haemostasis and Thrombosis
Vol. 7 Physico-Chemical Aspects of Drug Action
Vol. 8 Salt and Water Balance
Vol. 9 Pharmacology and Pain
Vol. 10 Rapporst Entre les Actions Pharmacologiques des LM.A.O. et Leurs Effets
chez V Homme
Vol. 11 Immunopharmacology
Immunopharmacology
Edited by
H. O. SCHILD
University College
London
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PERGAMON PRESS
O X F O RD L O N D ON • E D I N B U R G H • N EW Y O RK
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Copyright © 1968
Pergamon Press Ltd.
First edition 1968
Library of Congress Catalog Card No 67-19416
08 003269 9
LISTOF AUTHORS
BENACERRAF, B. Department of Pathology,
New York University, School of Medicine
New York, N.Y., U.S.A.
BROCKLEHURST, W. E. Edinburgh University, Medical School,
Edinburgh, U.K.
BRODER, I. Department of Medicine and Pharmacology,
University of Toronto,
Toronto, Canada
COCHRANE, C. G. Scripps Clinic and Research Foundation Division
of Experimental Pathology,
La Jolla, California, U.S.A.
DAVID, J. R. Department of Medicine, New York University School
of Medicine,
New York, N.Y., U.S.A.
FRAY, ANTOINETTE Chaire de Medecine Experimentale,
College de France,
Paris, France
GOTH, A. Department of Pharmacology, The University of
Texas, Southwestern Medical School,
Dallas, Texas, U.S.A.
GREAVES, M. W. Department of Pharmacology, University College,
London, England
HALPERN, B. Chaire de Medecine Experimentale,
College de France,
Paris, France
HAYASHI, H. Department of Pathology, Kumamoto University
Medical School,
Kumamoto, Japan
ISHIZAKA, K. Children's Asthma Research Institute and Hospital,
Denver, Colorado, U.S.A.
LEVINE, B. B. Department of Medicine, New York University School
of Medicine,
New York, N.Y., U.S.A.
MONGAR, J. L. Department of Pharmacology, University College,
London, England
M 0 LLER-EBERHARD, J. Scripps Clinic and Research Foundation, Division of
Experimental Pathology,
La Jolla, California, U.S.A.
vii
viii LIST OF AUTHORS
OVARY, Z. Department of Pathology, New York University
School of Medicine,
New York, N.Y., U.S.A.
ROTSCHILD, A. M. Department of Pharmacology,
Faculty of Medicine,
Ribeirao Preto, Brazil
SCHILD, H. O. Department of Pharmacology,
University College,
London, England
STORB, URSULA Chaire de Medecine Experimental,
College de France,
Paris, France
UNGAR, G. Baylor University, College of Medicine,
Houston, Texas, U.S.A.
UVNAS, B. Department of Pharmacology,
Karolinska Institutet,
Stockholm, Sweden
WARD, P. A. Scripps Clinic and Research Foundation,
Division of Experimental Pathology,
La Jolla, California, U.S.A.
WlLLOUGHBY, D. A. Department of Pathology,
St. Bartholomew's Hospital,
London, England
PREFACE
THIS volume contains the proceedings of a symposium on Immunophar-
macology held at Sao Paulo, Brazil, on 26 July 1966, in conjunction with
the Illrd International Pharmacological Congress.
The symposium expresses the close relations between pharmacology
and immunology, relations which are likely to become even closer as the
nature of the substances mediating hypersensitivity reactions gets further
elucidated.
Part I of this volume deals with immunoglobulins responsible for hyper-
sensitivity reactions and with the mechanisms of these reactions; part
II with pharmacological mediators of immediate and delayed hypersensi-
tivity and Arthus reactions and with soluble factors released by the action
of antigen on sensitized lymphocytes. Part III contains a single paper
on the subject of penicillin allergy.
I have been entrusted with the organization of the symposium together
with Dr. Benacerraf and am most grateful to the participants for their lucid
contributions and for providing the manuscripts in good time. I also wish
to thank Dr. Radan Capek, Prague, for helping with the editing and print-
ing of the volume.
H. O. SCHILD
ix
PROPERTIES OF IMMUNOGLOBULINS
WHICH MEDIATE THE RELEASE
OF VASOACTIVE AMINES IN
EXPERIMENTAL ANIMALS*
BARUJ BENACERRAF
Department of Pathology
New York University School of Medicine
New York, N.Y.
RECENT studies on the heterogeneity of immunoglobulins have revealed
that several mammalian species including man produce a special immuno-
globulin capable of sensitizing host tissues for systemic, local, and under
certain experimental circumstances, in vitro anaphylactic reactions. This
immunoglobulin type will be referred to as "anaphylactic antibody". Its
biological properties are a consequence of its capacity to combine with
certain target cells (probably tissue mast cells) so that subsequent contact
with a specific antigen initiates a series of events which cause the release
of vasoactive agents. The anaphylactic antibodies are characteristic of the
species in which they have been produced since they can only mediate
and transfer anaphylactic reaction within this species or to closely related
species. It is a general observation that anaphylactic antibodies cannot
sensitize unrelated species. However, mammalian anaphylactic antibodies
can nevertheless be classified into main types according to their physico-
chemical properties and some of thei(r 1b)io2logical pro perti()es.3 T,he 4anaphy-
lactic antibodies of the guinea pig ' and mouse are very similar
and have been called y\ immunoglobulin(s. 5)Th,ey6 diff(er) 7app rec)ia(bl8y from () 9
the anaphylactic antibodies of the rat, rabbit, dog and man
which will be referred to as the "reaginic type" of anaphylactic antibody.
* This work was supported by United Public Health Service Grants AI 2094 and
AI 04983 and by the Health Research Council of the City of New York under contract
no. 1-138.
3
4 B. BENACERRAF
I. ANAPHYLACTIC ANTIBODIES OF THE GUINEA PIG AND
MOUSE, L ITMMUNOGLOBULINS
A. The guinea pig.—Although the guinea pig has been an animal of
choice for the study of local and systemic anaphylactic reactions ever
since the discovery of these phenomena, characterization of the specific
guinea pig immunoglobulin which mediates these reactions has only been
accomplished in the past few years. This has been achieved independently
by two groups of workers in the United States and in England. Benacerraf,
Ovary, and Bloch observed that antisera of guinea pigs immunized with
hapten-protein conjugates contain two populations of prec1i, pit)at1ing0 anti-
bodies directed against the same antigenic determinant/ These two
antibodies differ in their electrophoretic mobility on agar gel and starch
block electrophoresis. They were identified as yi and y 2 antibodies. White,
Jenkins and Wilkinson made similar observations with antisera of g(u)i2nea
pigs immunized with egg albumin in complete Freund's adjuvant. Both
groups reported that the faster migrating y x but not y2 antibodies are
able to sensitize guinea pigs for passive cutaneous anaphylaxis (PCA)
and for systemic anaphylaxis. y 2 antibodies are able to block specifically
PCA reactions provoked by y± antibodies with the same immunological
specificity provided a sufficient excess is used. Contrasting with their ability
to mediate anaphylactic reactions, y± guinea-pig antibodies are not able
to fix complement in the presence of antigen, nor to sensitize antigen coated
erythrocytes for lysis by complement. Complement fixation was shown
to be a property of the slower migrating y 2 immunoglobulins.These
two antibo(d)y1 types were shown to have approximately 7S sedimentation
constants. Immunological analysis of y x and y2 guinea-pig antibodies
with rabbit antisera against these immunoglobulins and their papain-
fragments revealed that y x and y2 antibodies are identical with respect
to their L chains and their Fd fragments, which c(on1 st2i tu1t)e 31the4 Fab frag-
ment containing the antibody combining site. ' ' However, these
two immunoglobulins differ completely in the portion of their H chains
contributing to the Fc fragments. These structural differences are not sur-
prising and contribute to explain the different biological properties of
these two antibody classes, since it has been clearly demonstrated that
the chemical structures respon(1s5ib)le for biological activities of antibodies, ( )1 6
such as complement fixation, and passive anaphylactic sensitization,
are indeed located on the Fc fragment of 7S immunoglobulins.
Although the level of yi immunoglobulins is usually much lower in the
serum of unimmunized guinea pigs than the level of y 2 immunoglobulins,
PROPERTIES OF IMMUNOGLOBULINS 5
specific immu(n)liz7ation will cause a very high level of y± antibodies to be
synthesized. Immunization with protein antigens, in complete adju-
vants containing mycobacteria, st(im 1u)la2tes the synthesis of comparable
quantities of y2 and y\ antibodies ' although the respective amount of
these two imm(un)1og8lobulins formed differs somewhat with the type of
antigens used. Immunization without adjuv(a)1nts stimulates the forma-
tion of yi immunoglobulin almost exclusively. It can be stated, therefore,
that anaphylactic antibody of the guinea pig is one of its two most preva-
lent immunoglobulins and for this reason it has been possible to obtain
it in sufficient amounts, relatively pure, to allow its characterization and
the study of its physical properties.
The anaphylactic properties of guinea pig y± antibodies whether in th(e 12)9 0
serum or purified are unaffected by heating to 56°C from 30 min to 4 fir, '
a treatment which is known to (d)e7stroy the activity of human, rat and
rabbit anaphylactic antibodies. Alkylation and reduction under con(- )2 1
ditions known to completely inactivate human anaphylactic antibodies
reduced the skin sensitizing activity of guinea pig purified y± antihapten
antibodies only slightly. Because of their important differences in electric
charge, guinea pig y± and y 2 immunoglobulins can be easily separated
either by preparat(iv)2e 2zone electrophoresis^ or by chromatography on
DEAE cellulose.
In order to differentiate guinea pig y± immunoglobulins from the IgA ( )2 3
antibodies, which are characterized by a high content of carbohydrates,
hexose analysis of both y x and y2 (g)u2in2ea pig antibodies were made and
these were found to be identical. Since the placental membranes of
some animals select certain maternal antibodies for transmission to the
fetus and exclude others, passage of y\ and y 2 immunoglobulins from
mother to fetus was investigated. Both immunoglobulin types are trans-
mitted to the young by active(ly )2or0 passively immunized pregnant moth-
ers in comparable amounts. Thus the site on the Fc fragment of these
two immunoglobulin types which is involved in placental membrane
transport in this species is present in both types of guinea pig antibodies.
B. The mouse.—The immunoglobulins of the mouse have been well
investigated and the following types have bee(n )i2de4ntified: IgM, IgA, and
two IgG's also referred to as y 2A and y2B. Besides (th)2ese5 immunoglo-
bulins, the mouse produces also a y± immunoglobulin which appears
to b e (2id3 e4n)t2ica5l in many respects with the guinea pig y± immunoglobu-
lin. ' ' The y1 mouse antibodies have a faster electrophoretic mobility
than mouse y2 antibodies, although this difference is not as marked as in
the guinea pig, which renders the separation of yi from y 2 mouse immu-
6 B. BENACERRAF
noglobulins more difficult. Mouse y± antibodies have been shown to be
the only immunoglobulin class capable of transferring passive cutaneous
anaphylaxis in the mouse and can be consi3d, e)re4d therefore to be the ana-
phylactic immunoglobulin of the species/ Mouse y± is also a 7S immu-
noglobulin. It is found in the serum in lesser amounts than y 2 immuno-
globulins but it is synthesized in large quantities following specific immu-
nization. Similarly to guinea pig y± immunoglobulins, mouse anaphylactic
antibodies appea(r) 3not to lyse antigen coated erythrocytes in the presence
of complement. Mouse y x immunoglobulins are synthesized in sufficient
amounts to be characterized and to allow their physicochemical properties
to be investigated. The anaphylactic antibody of the mouse is somewhat
more sensitive than that of the guinea p(ig) 3 to reduction and alkylation
but equally resistant to heat at 56°C.
In summary both mouse and guinea pig produce in large amounts a
7S immunoglobulin type with somewhat faster mobility than their re-
spective 72 antibodies. These y± globulins possess distinct antigenic deter-
minants on the Fc fragment of their respective H chains. They mediate
anaphylactic reactions in their respective species. They are also similar
in some of their biological characteristics. Both require a relatively short
latent period for passive cuta(n)2eo6us anaphylactic sensitization(, )2v7arying
from one hour in the mouse, to 3 to 5 hr in the guinea pig and they
bind to the skin for a very short period of time. Greatly decreased reac-
tions( a2 re 6)o2bs7erved after 24 hr in the mouse and after 48 hr in the guinea
pig. ' These sensitizing characteristics are quite different from those
of the anaphylactic antibodies of the rat, the rabbit, the dog and man
which bind much more strongly to the skin of their respective species.
They are the result of a comparatively lesser affinity of the mouse and
guinea pig 71, anaphylactic antibodies for their respective target cells.
In spite of their numerous similarities, the y x immunoglobulins of the
guinea pig and the mouse are characteristic of their respective species
as neither species can be sensitized by the yi antibodies of the other
species.
II. ANAPHYLACTIC ANTIBODIES OF THE RAT, THE RABBIT AND
THE DOG, "REAGENIC TYPE"
The anaphylactic antibodies of these three species are very similar and
resemble closely in their physicochemical and biological properties the
human reagin. They can be considered therefore to belong to a distinct
immunoglobulin class.
