A Historical Perspective on Evidence-Based Immunology

367 Pages·2015·27.356 MB·English

by  Moticka, Edward J

Dedication
This book is dedicated to

• Students, past, present, and future; and
• My wife, Jane Adrian, who provided encouragement, enthusiastic support, and confidence in this project.
Without her the book would never have been completed.
A HISTORICAL
PERSPECTIVE ON
EVIDENCE-BASED
IMMUNOLOGY
E J. M , P d
dward oticka h
Professor and Chair, Basic Medical Sciences
School of Osteopathic Medicine in Arizona
A.T. Still University
Mesa, AZ, USA
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Foreword
Students and others initiating the study of immunol- • The discovery by two hematology fellows,
ogy are confronted with numerous details about the William Harrington and James Hollingsworth,
immune system and immune responses that need to be that idiopathic thrombocytopenia purpura is an
assimilated into their knowledge base. These details are autoimmune disorder produced by antibodies
currently accepted by the community of immunologists; specific for the patient’s platelets.
however, upon initial publication, the experiments and • Georges Köhler was a postdoctoral fellow in César
supporting data often engendered controversy. Exam- Milstein’s laboratory when these two scientists
ples include the notion that the lymphocyte is the pri- developed the technique leading to the production
mary immunocompetent cell, the validity of the clonal of monoclonal antibodies.
selection theory and its displacement of instruction theo- • Immunology is a young discipline. While anecdotal
ries, the role of central lymphoid organs (thymus, bursa evidence existed for millennia that recovery from
of Fabricius, bone marrow) in maturation of immuno- an infectious disease protects an individual from
competent B and T lymphocytes, and the requirement subsequent development of the same disease, the
for cell interactions in the initiation of effective adaptive study of immunology as a scientific and clinical
immune responses. Without some knowledge of the discipline dates from the late eighteenth century.
background to these facts, the student misses out on the • The reach of immunology into medicine has evolved
rich history and compelling stories that bring immunol- from attempts to prevent infectious disease to a
ogy to life. It is to provide a sample of these stories that discipline that is intimately involved in virtually
A Historical Perspective on Evidence Based Immunology was every aspect of contemporary medicine.

written.
The idea for this book had a long gestation. As a
Several realities about immunology and immunologi-
graduate student, I enrolled in an immunochemistry
cal research emerged during the preparation of this book:
course taught by Alfred Nisonoff at the University of
• Immunology is an international endeavor. Scientists Illinois, Chicago. His approach to teaching included
and clinicians from six of the seven continents reading the primary literature, discussing the experi-
performed experiments and observations that are ments performed and the conclusions reached, and
included in this volume. determining what might be the next experiment to pur-
• Students and postdoctoral fellows produce a sue. This course took place in the late 1960s shortly after
significant number of findings including the the establishment of the basic structure of the immu-
following: noglobulin molecule. The journal articles read in this
• George Nuttall’s description of a serum substance course led eventually to division of the heavy and light
(antibody) induced in rabbits injected with Bacillus chains of immunoglobulin into constant and variable
anthracis that killed the bacteria. At the time Nut- regions. This, in turn, was critical for determining the
tall was a medical student in Germany. genetic makeup of the molecule and the mechanisms
• Jacques Miller’s discovery, shortly after receiving responsible for generation of diversity of both immu-
his PhD, that the thymus plays a critical role in the noglobulins and T cell receptors.
maturation of lymphocytes responsible for fight- In 2011, my wife and I visited the Walter and Eliza
ing infections. Hall Institute for Medical Research in Melbourne, Aus-
• Bruce Glick’s observation during his graduate tralia, where we spent a fabulous afternoon discuss-
training that the bursa of Fabricius in chickens is ing immunology with Jacques Miller. Following this
required for the maturation of antibody-forming experience, the desire to proceed with this volume was
lymphocytes. reenforced.
• Don Mosier’s experiments while a medical student In addition to Drs Nisonoff and Miller, I am indebted to
demonstrating that optimal antibody production several other individuals who provided encouragement for
requires both plastic adherent cells (macrophages) the project and/or read various chapters prior to publica-
and plastic nonadherent cells (lymphocytes). tion. These include J. John Cohen, MD; David Scott, PhD;
xi
xii FOREWORD
Max Cooper, MD, PhD; Katherine Knight, PhD; Jay and commented on virtually every chapter. Our
Crutchfield, MD; Sharon Obadia, DO; Robin Pettit, PhD; discussions improved the accuracy of the information
Milton Pong, PhD; and Katherine Brown, PhD. I also contained although any errors of fact or omission are
thank the deans at A.T. Still University including Drs Doug the authors alone,
Wood, Thomas McWilliams, Kay Kalousek, and Jeffrey • the editors, Mary Preap, Julia Haynes, and
Morgan who provided me the time to pursue this activity. Linda Versteeg-Buschman for their patience and
Other individuals critical to the successful completion encouragement, and
of this project include the following: • my wife, Jane Adrian, EdM, MPH. Jane read the
entire manuscript several times and we discussed
• the librarians at Arizona State University and A.T. it extensively. During these discussions, she
Still University particularly Catherine Ryczek who advocated for students and encouraged clarity in the
tirelessly filled my numerous requests for copies of description of the experiments and the interpretation
journal articles from both the United States and the of their results. Without her scientific expertise as a
rest of the world, clinical laboratory scientist, her skill as an educator,
• David Gardner, PhD, geneticist/molecular biologist, and her experience as a published author, this book
a colleague and a good friend who patiently read would not have been possible.
Glossary of Historical Terms
Investigators often assigned unique names for identical Helper peak 1 (HP-1)—IL-1
structures or molecules. This dichotomy of terms is confusing
Hepatocyte-stimulating factor—IL-1
for students as they read some of the older literature. To assist
Horror autotoxicus—a hypothesis proposed by Paul Ehrlich
in understanding these older terms, this glossary provides a
that the immune system was incapable of producing patho-
list of several of these terms with contemporary equivalents.
logical reactions to self (autoimmune disease)
19S gamma globulin—IgM
Hybridoma growth factor—IL-6
7S gamma globulin—IgG
Immunokörper—immune body—German term used for
Alexin—an original term for complement antibody
Amboceptor—an original term for an antibody that bound to Interferon β-2—one of the original designations of IL-6
a pathogen and to complement (alexin) thereby destroying
I —immune response gene(s); genes to which immune response
the pathogen R
are linked; counterpart of class II genes
Arthus reaction—a skin reaction originally induced by repeated
I —immune suppressor genes; genes thought to code for sup-
injections of horse serum into rabbit skin. The skin reaction S
pressive factors synthesized and secreted by T suppressor
is due to formation of antigen–antibody complexes that
lymphocytes
activate the complement system and induce inflammation.
Killer cell helper factor—IL-2
B cell-activating factor (BAF)—name given to a culture super-
Ly antigens—antigens expressed on mouse lymphocytes used
natant that activated B lymphocytes in vitro: IL-1
to develop polyclonal antibodies allowing characterization
B cell-differentiating factor (BCDF)—a factor in culture super-
of subpopulations of T lymphocytes
natants that induces antibody synthesis but not mitosis in B
Lymphocyte-activating factor (LAF)—IL-1
lymphocytes: IL-6
Pfeiffer phenomenon—the killing of Vibrio cholerae in the guinea
B cell growth factor—a factor in culture supernatants that
pig peritoneal cavity when the microbe is injected along with
induces mitosis in B lymphocytes: IL-4
antibody specific for V. cholerae. An early demonstration of
B cell-stimulating factor 1—IL-4
complement activity.
B cell-stimulating factor 2—IL-6
Phylocytase—antibody
Cluster of differentiation (CD)—a system of nomenclature for
Prausnitz-Küstner (P-K) reaction—demonstration of type I
molecules expressed primarily on peripheral blood white
(IgE-mediated) hypersensitivity induced by passive transfer
blood cells originally devised by an international workshop
of serum from an allergic to a nonallergic individual.
on Human Leukocyte Differentiation Antigens. Initially it
Reagin—term used to describe the antibody responsible for
was used to classify monoclonal antibodies produced by
type I hypersensitivity; IgE
different laboratories. Over 300 different CD markers are
currently recognized. Schick test—a skin test devised to determine if a patient has
sufficient antibody to protect against infection with
Copula—something that connects; used to refer to the molecule
Corynebacterium diphtheriae
that connects a pathogen with complement—antibody
Schultz–Dale reaction—in vitro assay to study type I hypersen-
Costimulator—an early term for antibody
sitivity. Uterine smooth muscle removed from a sensitized
CTLA4—cytotoxic T lymphocyte antigen 4; CD152
guinea pig is exposed in vitro to the sensitizing antigen.
Desmon—an early term for antibody The amount of muscle contraction is proportional to the
Dick test—a skin test used to determine if an individual is degree of sensitization.
immune to scarlet fever. Toxin from a culture of Streptococcus Secondary T cell-inducing factor—IL-2
pyogenes is injected intradermally. A positive test, character-
Substance sensibilisatrice—antibody
ized by an erythematous reaction within 24 h, indicates the
T4—antigen expressed by helper lymphocytes; now CD4
individual is not immune to the pathogen.
T8—antigen expressed by cytotoxic lymphocytes; now CD8.
Fixateur—a substance (antibody) that connects a pathogen with
complement T cell growth factor (TCGF)—IL-2
xiii
xiv GLOSSARY OF HISTORICAL TERMS
T cell-replacing factor—IL-1 Zwischenkörper—“between body”; antibody
T cell-replacing factor 3 (TRF-III)—IL-1 β2A—original definition of IgA antibody based on electrophoretic
T cell-replacing factor-μ—IL-1 mobility
T lymphocyte mitogenic factor—IL-2 γ-globulin—IgG
Thymocyte-stimulating factor (TSF)—IL-2 γ-M—IgM
C H A P T E R
1
Innate Host Defense Mechanisms and Adaptive
Immune Responses
O U T L I N E
Introduction 1 Adaptive Immune Responses 5
Anatomy 5
Innate Defense Mechanisms 1
Lymphocytes of the Adaptive Immune
Anatomy 2
Response 6
Cells of the Innate Host Defenses 2
Effector Mechanisms 6
Antimicrobial Molecules 3
Recognition of Pathogens 7
Effector Mechanisms 3
Inflammation 3 Conclusion 7
Phagocytosis 4
References 7
Complement 4
NK Lymphocyte-mediated Cytotoxicity 4 Time Line 8
Recognition of Pathogens 4
INTRODUCTION system to arise comprises innate or naturally occurring
mechanisms. The components of this system are found in
All multicellular life forms, including plants, inverte- plants, invertebrates, and vertebrates. The second system,
brates, and vertebrates, have devised defense strategies the adaptive immune response, evolved in vertebrates
that permit individuals to lead a healthy, relatively dis- after divergence from the invertebrate lineage, about
ease-free life. Knowledge about the mechanisms that have 500 million years ago. Interactions between the innate
evolved to protect humans derives initially from anec- host defenses and the adaptive immune responses are
dotal evidence that recovery from diseases such as small- generally successful in eliminating potential pathogens.
pox or the plague protects the individual from developing This chapter compares innate host defenses with
the same disease a second time. The acceptance of Louis adaptive immune responses as they function indepen-
Pasteur’s germ theory of disease in the mid-nineteenth dently and interdependently to eliminate potential
century resulted in the concept of an immune response pathogens. The chapter reviews the historical evidence
whose function is to provide this protection. Over the that provides the foundation for understanding the
ensuing 150 years, many studies have addressed how immune system and how the defense mechanisms at
our bodies deal with both pathogenic and nonpathogenic times defend us and at other times harm us.
microbes in our environment. Analysis of these mecha-
nisms, and the ability to manipulate them to our advan-
tage, constitutes the discipline of immunology. INNATE DEFENSE MECHANISMS
Two separate but interrelated host defense systems
have evolved to defend the individual from attack by Most potential pathogens are defeated by innate host
potential pathogens. In this text, pathogen is used in its defense mechanisms. Innate host defenses include phys-
broadest sense to refer to any external agent that can ical barriers such as the skin and the mucous membranes
cause disease (pathology). Evolutionarily the first defense along the gastrointestinal, respiratory, and genitourinary
1
A Historical Perspective on Evidence-Based Immunology
http://dx.doi.org/10.1016/B978-0-12-398381-7.00001-0 © 2016 Elsevier Inc. All rights reserved.
2 1. INNATE AND ADAPTIVE IMMUNITY
tracts, nonspecific cells such as macrophages and granu- performed by these structures remained unappreciated
locytes, molecules including mediators of inflammation until general acceptance of the germ theory of disease
and proteins of the complement system, and effector in the second half of the nineteenth century. The devel-
mechanisms such as phagocytosis and inflammation. opment of the germ theory is generally credited to John
Recognition of a pathogen by the cells of this innate Snow (1813–1858) who in 1849 studied an outbreak of
defense system results in the release of an array of anti- cholera in London and traced it to a water well on Broad
microbial molecules, such as lysozyme and defensins Street. Experimental proof of the germ theory was pro-
into the local environment. These molecules kill a vari- vided by Louis Pasteur (1822–1895). He demonstrated
ety of pathogenic microorganisms and are involved in that microbes were responsible for fermentation of beer
enhancing ongoing inflammatory responses, a major and wine as well as spoilage of beverages such as milk.
effector mechanism of the innate system. He extended these observations to reveal that human
Innate host defense mechanisms and adaptive and animal diseases could also be caused by microbes
immune responses differ in three important characteris- (Pasteur, 1880). Once the ubiquity of microorganisms
tics in their response to pathogens: was recognized, the interaction between the skin and

mucous membranes with the environment became an
• Cells of the innate host defenses are poised to
area of biological research.
respond immediately while the cells of the adaptive
The presence of cilia on mucous membranes provides
immune response require activation.
an additional barrier to the breaching of these surfaces
• Innate host defense mechanisms are not specific
by pathogens. Cilia and the presence of mucous enhance
while adaptive immune responses produce cells and
the protective function of these barriers by increasing the
molecules that are highly specific for and target the
challenge for microbes attaching to and penetrating these
pathogen.
membranes. Several antimicrobial substances, including
• Innate host defense mechanisms lack memory of
lysozyme, phospholipase-A, and defensins, are found
past responses should the host be invaded a second
in secretions on these physical barriers. Lysozyme and
time by the same pathogen while adaptive immune
phospholipase-A are present in tears, saliva, and nasal
responses display memory by mounting a more
secretions while defensins and lysozyme are present
rapid response, resulting in an increased number of
along the mucous membranes lining the respiratory and
specific lymphocytes and a higher titer of antibodies
gastrointestinal tracts.
to a second exposure.

Cells of the Innate Host Defenses
Inflammation and phagocytosis are the two primary
effector mechanisms by which the innate host defense Three cell types provide protection against potential
system eliminates pathogens. Macrophages, a major pathogens in the innate host defense system:

phagocytic cell, migrate throughout the body, recog-
• granulocytes, including neutrophils, basophils, and
nizing and engulfing foreign material. Phagocytosis,
eosinophils;
the ingestion of solid particles such as microorganisms,
• phagocytic cells, including monocytes, macrophages,
induces gene transcription in the phagocytes, resulting in
and dendritic cells; and
the synthesis and secretion of mediators of the inflamma-
• a subset of lymphocytes with natural cytotoxicity
tory response such as cytokines and chemokines. Inflam-
potential.
mation recruits other cells into the local environment to
play a role in eliminating the pathogen. These cells, classified as leukocytes, are found in the
Innate host defense mechanisms depend on the pres- peripheral blood and distributed throughout the organs
ence of certain anatomical structures and cells, effector of the body. The initial morphological descriptions of
mechanisms, and recognition structures. In the follow- leukocytes appeared in the 1840s when Gabriel Andral
ing sections the history of each of these components is in France and William Addison in England reported
reviewed. It is noted when the historical background of the presence of white cells in peripheral blood (Hajdu,
a particular subject is covered in subsequent chapters of 2003). These observations were followed by reports of
this book. increased numbers of peripheral blood leukocytes that
could be correlated with various diseases, including
tuberculosis and sexually transmitted infections. In 1845
Anatomy
Rudolph Virchow (1821–1902) in Germany and John
The main anatomical components of the innate host Hughes Bennett (1812–1875) in Scotland simultaneously
defense mechanisms include the skin and the mucous described the peripheral blood cells of patients with leu-
membranes lining the respiratory, gastrointestinal, and kemia (Chapter 35).
genitourinary tracts. These structures provide a barrier The functions of the cells of the innate host defense
to invasion of the body by pathogens. The protective role system became the focus of studies for the remainder of
INNATE DEfENsE MECHANIsMs 3
the nineteenth century. Two cell types, macrophages and with coryza (the common cold), he tried to isolate and
granulocytes, are primarily involved in the removal of culture a causative agent from the individual’s nasal
invading pathogens by the innate defense mechanisms. secretions. He was unsuccessful until day 4 when he
In 1879, Paul Ehrlich (1854–1915) initially described noted growth of small colonies of large, gram-positive
granulocytes based on staining characteristics using diplococcus that he termed Micrococcus lysodeikticus.
dyes he developed in his laboratory. Ilya Metchnikov This bacterium is now classified as Micrococcus luteus
(also Elie Metchnikoff) (1845–1916) provided descrip- and is recognized as part of the normal flora. Applica-
tions of macrophages and developed his “phagocytic tion of a saline extract of nasal mucosa to cultures of
theory of immunity” in 1884 (Chapter 15). M. luteus produced lysis of the bacteria. Lysozyme, as
In addition to macrophages and granulocytes, a third this extract is called, is present in many bodily fluids and
cell type, the NK (natural killer) lymphocyte, is consid- tissues. Lysozyme is now known to provide protection
ered a component of the innate host defenses. NK cells against several gram-positive bacteria, especially on the
are a heterogenous population of lymphocytes charac- conjunctiva of the eye and along mucous membranes.
terized by their ability to lyse various cellular targets, Fleming received his early schooling in Scotland. In
particularly malignant cells and cells infected with a 1906 he was awarded the MBBS (MD) degree from St
variety of intracellular pathogens. They were discovered Mary’s Hospital Medical School in London. He served
in the early 1970s based on the destruction of tumor cells. as an assistant to Sir Almroth Wright (discoverer of com-
Morphologically, many of these cells are large granular plement—Chapter 12) at St Mary’s and as an instructor
lymphocytes. NK lymphocytes exist in mice, humans, in the medical school. Following service in World War
and other vertebrates. The experiments that character- I (1914–1918) Fleming returned to London to assume a
ized these cells are presented in Chapter 28. professorship at the University of London.
Fleming is best known for his discovery of penicillin
Antimicrobial Molecules in 1929 when a fungus contaminated a culture of Staph-
In 1894, A.A. Kanthak and W.B. Hardy, working at ylococcus while he was away from his laboratory. He
Bartholomew Hospital in London and at Cambridge, returned from his summer holiday to find that the fun-
injected rats and guinea pigs intraperitoneally with gus had secreted a substance that inhibited the growth
Bacillus anthracis, Pseudomonas aeruginosa, or Vibrio chol- of Staphylococcus as well as other gram-positive bacteria.
erae. At intervals they killed the animals, removed cells Fleming was unsuccessful in purifying this inhibitory
from their peritoneal cavities, and examined with a substance; however, Howard Florey (1898–1968) and
microscope. Kanthak and Hardy observed that granulo- Ernst Boris Chain (1906–1979) succeeded and developed
cytes surrounded the bacteria and extruded their gran- the fungal metabolite into the important antimicrobial
ules upon contact while macrophages phagocytized drug, penicillin. Fleming, Florey, and Chain shared the
the microbes. Those bacteria that were contacted by the Nobel Prize in Physiology or Medicine in 1945 “for the
granulocytes were destroyed. One conclusion from this discovery of penicillin and its curative effect in various
study was that the released granules must contain anti- infectious diseases.”
microbial substances.
Numerous investigators attempted to characterize this
Effector Mechanisms
antimicrobial material but were unsuccessful for more
than 70 years. In 1966, H.I. Zeya and John Spitznagel
In immunological terms, effector mechanisms refer
at the University of North Carolina (1966a,b) isolated
to the cells and/or molecules that are activated through
the contents of the granules using electrophoresis. They
interaction with a pathogen and subsequently inhibit the
demonstrated that the antibacterial activity was found
pathogen from causing disease. The innate host defenses
in at least three separate molecules. In 1984, Mark
employ four effector mechanisms:
Selsted and colleagues at the University of California,
Los Angeles purified the active material from rabbit • inflammation,
granulocytes and demonstrated that it consisted of a • phagocytosis,
group of molecules they termed defensins. Defensins are • complement activation, and
low molecular weight peptides that have antimicrobial • cell-mediated cytotoxicity.
activity. They are produced and stored in granulocytes
Inflammation
of the peripheral blood and the Paneth cells of the intes-
tine. Defensins are also found on the skin and along the More than 2400 years ago, Hippocrates developed a
mucous membranes of the respiratory, genitourinary, theory of the four cardinal humors to explain disease.
and gastrointestinal tracts. These four humors, blood, phlegm, choler (yellow bile),
In 1922, Alexander Fleming (1881–1955) described and melancholy (black bile), needed to be in balance
lysozyme (muramidase). While studying an individual for a person to be healthy. Many disease treatments