INTRODUCTION TO BASIC IMMUNOLOGY

MLS 326
BY
MR. OYAKHIRE FIDELIS
JOSEPH AYO BABALOLA UNIVERSITY (JABU), IKEJI-ARAKEJI, OSUN STATE
DEPARTMENT OF MEDICAL LABORATORY SCIENCE,
COLLEGE OF HEALTH SCIENCES

HISTORICAL OVERVIEW OF IMMUNOLOGY

The fundamental observation that led to the development of immunology as a scientific discipline
was that an individual might become resistant for life to a certain disease after having contracted it
only once. The term immunity, derived from the Latin immunis (exempt), was adopted to designate
this naturally acquired protection against diseases such as measles or smallpox. The term immunity is
derived from the Latin word “immunis” (exempt), which was originally referred to the protection
from legal prosecution offered to the Roman senators during their tenures in office. This term was
adopted subsequently to designate the naturally acquired protection against diseases, such as measles
or smallpox. It indicated that an individual can develop lifelong resistance to a certain disease after
having contracted it only once. The cells and molecules responsible for immunity constitute the
immune system, and their collective and coordinated response to foreign substances is called the
immune response
The emergence of immunology as a discipline was closely tied to the development of microbiology.
The work of Pasteur, Koch, Metchnikoff, and many other pioneers of the golden age of microbiology
resulted in the rapid identification of new infectious agents. This was closely followed by the
discovery that infectious diseases could be prevented by exposure to killed or attenuated organisms or
to compounds extracted from the infectious agents. The impact of immunization against infectious
diseases such as tetanus, measles, mumps, poliomyelitis, and smallpox, to name just a few examples,
can be grasped when we reflect on the fact that these diseases, which were at one time significant
causes of mortality and morbidity, are now either extinct or very rarely seen. Indeed, it is fair to state
that the impact of vaccination and sanitation on the welfare and life expectancy of humans has had no
parallel in any other developments of medical science.
In the second part of this century immunology started to transcend its early boundaries and become a
more general biomedical discipline. Today, the study of immunological defense mechanisms is still
an important area of research, but immunologists are involved in a much wider array of problems,
such as self-nonself discrimination, control of cell and tissue differentiation, transplantation, cancer
immunotherapy, etc. The focus of interest has shifted toward the basic understanding of how the
immune system works in the hope that this insight will allow novel approaches to its manipulation.
The concept of immunity has existed since the ancient times. An example is the Chinese custom of
making children resistant to smallpox by making them inhale powders made from the skin lesions of
patients recovering from the disease. The first European mention of immunity is recorded by
Thucydides in Athens during the fifth century BC. In describing plague in Athens, he wrote in 430
BC that only those who had recovered from plague could nurse the sick, because they would not
contract the disease a second time. Once the concept of existence of immunity was established, it was
not long before manipulation of immunity under controlled conditions followed. First, it was Edward
Jenner who in a successful experiment injected the material from a cowpox pustule into the arm of an
8-year-old boy and demonstrated the lack of development of disease after subsequent exposure to
smallpox. This was based on his observations that milkmaids who had suffered from cowpox never
contracted the more serious smallpox. Jenner’s technique of inoculating with cowpox to protect
against smallpox spread quickly throughout Europe. However, for many reasons, including lack of
knowledge of obvious disease targets and their causes, it was after nearly hundred years that this
technique was applied to prevent smallpox. The experimental work of Emil von Behring and
Shibasaburo Kitasato in 1890 gave the first insights into the mechanism of immunity, earning von
Behring the Nobel Prize in Medicine in 1901. Von Behring and Kitasato demonstrated that serum
(the liquid, noncellular component of coagulated blood) from animals previously immunized to
diphtheria could transfer the immune state to unimmunized animals. Since then, immunology as a
field of study has come a long way. It has been and remains one of the hottest fields of research as
shown by the statistic that about 17 Nobel Prizes have been awarded to scientists involved in
immunological research
TYPES OF IMMUNITY
The main function of the immune system is to prevent or limit infections by pathogenic
microorganisms, such as bacteria, viruses, parasites, and fungi. The recognition of microorganisms
and foreign substances is the first event in immune responses of a host. The body’s defense
mechanisms can be divided into: (a) innate (natural) immunity and (b) acquired (adaptive) immunity.
INNATE IMMUNITY
Innate immunity is the resistance that an individual possesses by birth. Innate immunity may be
classified as (a) individual immunity, (b) racial immunity, and (c) species immunity.
Individual immunity: Individual immunity denotes resistance to infection, which varies within
different individuals in the same race and species and is genetically determined. For example, if one
homozygous twin develops tuberculosis, there is a very high possibility that the other twin will also
develop tuberculosis. But in heterozygous twins, there is a very low possibility of the other twin
suffering from tuberculosis.
Racial immunity: Racial immunity denotes a difference in susceptibility or resistance to infection
among different races within a same species. For example, races with sickle cell anemia prevalent in
Mediterranean coast are immune to infection caused by malaria parasite, Plasmodium falciparum.
This is due to a genetic abnormality of erythrocytes, resulting in sickleshaped erythrocytes that
prevent parasitization by P. falciparum. Similarly, individuals with a hereditary deficiency of
glucose6-phosphatase dehydrogenase are also less susceptible to infection by P. falciparum.
Species immunity: Species immunity denotes a total or relative resistance to a pathogen shown by all
members of a particular species. For example, chickens are resistant to Bacillus anthracis, rats are
resistant to Corynebacterium diphtheriae, whereas humans are susceptible to these bacteria. The exact
reason for such type of immunity is not known.
Key Points
Innate immunity shows the following features:
■ It is due to the genetic and constitutional makeup of an individual. Prior contact with
microorganisms or their products is not essential.
■ It acts as the first line of defense of the host immune system.
■ The mechanisms involved in innate immunity are present in place even before exposure to the
foreign agent. They are not specific to any infectious agent and do not seem to improve response on
repeated exposures.
■ Phagocytic cells (e.g., macrophages and neutrophils), barriers (e.g., skin and mucous membrane),
and a variety of antimicrobial compounds synthesized by the host, all play important roles in innate
immunity.
FACTORS INFLUENCING INNATE IMMUNITY
The factors that may influence innate immunity of the host include age and nutritional status of the
host.
Age: Extremes of age make an individual highly susceptible to various infections. This is explained
in part by the immature immune system in very young children and waning immunity in older
individuals. The fetus-in-utero is usually protected from maternal infections by the placental barrier.
However, human immunodeficiency virus (HIV), rubella virus, cytomegalovirus, and Toxoplasma
gondii cross the placental barrier and cause congenital infections. Very old people are susceptible to
suffer more than young people from a disease (e.g., pneumonia) and have high mortality. Measles,
mumps, poliomyelitis, and chicken pox are few examples of the diseases that cause more severe
clinical illness in adults than in young children. This may be due to more active immune response in
an adult causing greater tissue damage.
Nutritional status: Nutritional status of the host plays an important role in innate immunity. Both
humoral and cell mediated immunities are lowered in malnutrition. Examples are:
■ Neutrophil activity is reduced, interferon response is decreased, and C3 and factor B of the
complement are decreased in protein–calorie malnutrition.
■ Deficiency of vitamin A, vitamin C, and folic acid makes an individual highly susceptible to
infection by many microbial pathogens.
Hormonal levels: Individuals with certain hormonal disorders become increasingly susceptible to
infection. For example, individuals suffering from diabetes mellitus, hypothyroidism, and adrenal
dysfunction are increasingly susceptible to staphylococcal infection, streptococcal infection,
candidiasis, aspergillosis, zygomycosis and many other microbial infections. Similarly, pregnant
women are more susceptible to many infections due to higher level of steroid during pregnancy.
MECHANISMS OF INNATE IMMUNITY
Innate immunity of the host performs two most important functions: it kills invading microbes and it
activates acquired (adaptive) immune processes. Innate immunity unlike adaptive immunity,
however, does not have any memory and does not improve after re-exposure to the same
microorganism. The innate immunity is primarily dependent on four types of defensive barriers:
(a) anatomic barriers, (b) physiologic barriers, (c) phagocytosis, and (d) inflammatory responses.
Anatomic barriers: Anatomic barriers include skin and mucous membrane. They are the most
important components of innate immunity. They act as mechanical barriers and prevent entry of
microorganisms into the body. The intact skin prevents entry of microorganisms. For example, breaks
in the skin due to scratches, wounds, or abrasion cause infection. Bites of insects harboring
pathogenic organisms (e.g., mosquitoes, mites, ticks, fleas, and sandflies), introduce the pathogens
into the body and transmit the infection. Skin secretes sebum, which prevents growth of many
microorganisms. The sebum consists of lactic acid and fatty acids that maintain the pH of skin
between 3 and 5, and this pH inhibits the growth of most microorganisms. Mucous membranes form
a large part of outer covering of gastrointestinal,respiratory, genitourinary, and many other tracts of
human host. A number of nonspecific defense mechanisms act to prevent entry of microorganisms
through mucous membrane. ■ Saliva, tears, and mucous secretions tend to wash away potential
invading microorganisms, thereby preventing their attachment to the initial site of infections. These
secretions also contain antibacterial or antiviral substances that kill these pathogens. ■ Mucus is a
viscous fluid secreted by the epithelial cells of mucous membranes that entraps invading
microorganisms. ■ In lower respiratory tract, mucous membrane is covered by cilia, the hair-like
protrusions of the epithelial cell membranes. The synchronous movement of cilia propels
mucusentrapped microorganisms from these tracts. ■ In addition, nonpathogenic organisms tend to
colonize the epithelial cells of mucosal surfaces. These normal flora generally compete with
pathogens for attachment sites on the epithelial cell surface and for necessary nutrients.
Physiologic barriers: The physiologic barriers that contribute to innate immunity include the
following:
■ Gastric acidity is an innate physiologic barrier to infection because very few ingested
microorganisms can survive the low pH of stomach contents.
■ Lysozyme, interferon, and complement are some of the soluble mediators of innate immunity.
Lysozyme has antibacterial effect due to its action on the bacterial cell wall. Interferons are secreted
by cells in response to products of viral infected cells. These substances have a general antiviral effect
by preventing the synthesis of viral structural proteins. Complement is a group of serum-soluble
substances that when activated damage the cell membrane.
■ There are certain types of molecules that are unique to microbes and are never found in
multicellular organisms. The ability of the host to immediately recognize and combat invaders
displaying such molecules is a strong feature of innate immunity.
Phagocytosis: Phagocytosis is another important defense mechanism of the innate immunity.
Phagocytosis is a process of ingestion of extracellular particulate material by certain specialized cells,
such as blood monocytes, neutrophils, and tissue macrophages. It is a type of endocytosis in which
invading microorganisms present in the environment are ingested by the phagocytic cells. In this
process, plasma membrane of the cell expands around the particulate material, which may include
whole pathogenic microorganisms to form large vesicles called phagosomes.
Inflammatory responses: Tissue damage caused by a wound or by an invading pathogenic
microorganism induces a complex sequence of events, collectively known as the inflammatory
responses. The end result of inflammation may be the activation of a specific immune response to the
invasion or clearance of the invader by components of the innate immune system. The four cardinal
features of inflammatory responses are rubor (redness), calor (rise in temperature), dolor (pain), and
tumor (swelling). Mediators of inflammatory reactions: Histamine, kinins, acutephase proteins, and
defensin are the important mediators of inflammatory reactions.
■ Histamine: It is a chemical substance produced by a variety of cells in response to tissue injury. It is
one of the principal mediators of the inflammatory response. It binds to receptors on nearby
capillaries and venules, causing vasodilatation and increased permeability.
■ Kinins: These are other important mediators of inflammatory response. They are normally present
in blood plasma in an inactive form. Tissue injury activates these small peptides, which then cause
vasodilatation and increased permeability of capillaries. Bradykinin also stimulates pain receptors in
the skin. This effect probably serves a protective role because pain normally causes an individual to
protect the injured area.
■ Acute-phase proteins: These include C-reactive proteins and mannose-binding proteins that form
part of the innate immunity. These proteins are produced at an increased concentration in plasma
during acute-phase reaction, as a nonspecific response to microorganisms and other forms of tissue
injury. They are synthesized in the liver in response to cytokines called proinflammatory cytokines,
namely, interleukin-1 (IL-1), interleukin-6 (IL6), and tissue necrosis factor (TNF). They are called
proinflammatory cytokines because they enhance the inflammatory responses.
■ Defensins: They are another important component of the innate immunity. They are cationic
peptides that produce pores in membrane of the bacteria and thereby kill them. These are present
mainly in the lower respiratory tract and gastrointestinal tract. The respiratory tract contains
defensins, whereas the gastrointestinal tract contains defensins. The defensins also exhibit antiviral
activity. They bind to the CXCR4 receptors and block entry of HIV virus into the cell. How these
defensins differentiate microbes from some cells is not known.
ADAPTIVE (ACQUIRED) IMMUNITY
Adaptive immunity is also called acquired immunity, since the potency of immune response is
acquired by experience only.
KEY POINTS
Adaptive immunity shows the following features:
■ It is the resistance acquired by an individual during life.
■ It occurs after exposure to an agent and is mediated by antibodies as well as T lymphocytes (helper
T cells and cytotoxic T cells).
■ It has immunologic memory and a remarkable capability of discriminating between self and nonself
antigens.
■ Once an antigen has been recognized by the cells of acquired immune system, the response to it is
specific and can be repeated. In most cases, the acquired immune response improves with repeated
exposure.
■ The immune response to the second challenge occurs more quickly than the first, is stronger, and is
often more effective in neutralizing and clearing the pathogen.
TYPES OF ACQUIRED IMMUNITY
Acquired immunity against a microbe may be induced by the host’s response to the microbe or by
transfer of antibodies or lymphocytes specific for the microbes. It is of two types: active immunity
and passive immunity.

DIFFERENCES BETWEEN INNATE AND ADAPTIVE IMMUNITY

Features INNATE IMMUNITY ADAPTIVE IMMUNITY
Definition The resistance to infection that an The resistance that an individual
individual possesses by virtue of acquires during life
genetic and constitutional makeup
Types Nonspecific and specific Active and Passive
Time taken to develop Hours Days
Specificity For structures shared by groups of For antigens of microbes and for
related microbes nonmicrobial antigens
Memory None; repeated exposure brings Yes; secondary response much
response like primary response faster than primary response
Components
Physical and chemical Skin, mucosal epithelia, and Lymphocytes in epithelia and
barriers antimicrobial chemicals antibodies secreted at epithelial
cells
Blood and tissue Complement; leukins from Antibodies
antimicrobial substance leukocytes, plakins from platelets,
lactic acid found in muscle tissue,
lactoperoxidase in milk, and
interferons (antiviral
Cells Phagocytes (macrophages and Lymphocytes
neutrophils) and natural killer cell

◗ TYPES OF ACQUIRED IMMUNITY

Acquired immunity against a microbe may be induced by the host’s response to the microbe or by
transfer of antibodies or lymphocytes specific for the microbes. It is of two types: active immunity
and passive immunity.
ACTIVE IMMUNITY
The immunity induced by exposure to a foreign antigen is called active immunity. Active immunity is
the resistance developed by an individual after contact with foreign antigens, e.g., microorganisms.
This contact may be in the form of:
■ clinical or subclinical infection,
■ immunization with live or killed infectious agents or their antigens, or
■ exposure to microbial products, such as toxins and toxoids. In all these circumstances, the immune
system of the host is stimulated to elicit an immune response consisting of antibodies and activated
helper T (TH) cells and cytotoxic T lymphocytes/cells (CTLs). Active immunity develops after a
latent period, during which immunity of the host is geared up to act against the microorganism.
Hence it is slow in onset, especially during this primary response. However, once the active immunity
develops, it is long-lasting and this is the major advantage of the active immunity.
The active immunity is of two types: natural active immunity and artificial active immunity.
■ NATURAL ACTIVE IMMUNITY: It is acquired by natural clinical or subclinical infections.
Such natural immunity is longlasting. For example, individuals suffering from smallpox become
immune to second attack of the disease.
■ ARTIFICIAL ACTIVE IMMUNITY: It is induced in individuals by vaccines. There is a wide
range of vaccines available against many microbial pathogens. These may be live vaccines, killed
vaccines, or vaccines containing bacterial products.
Mediators of active immunity: Active immunity is mediated by humoral immunity and cell-mediated
immunity. These two types of immunities are mediated by different components of the immune
system and function in different ways to kill different types of pathogens

DIFFERENCES BETWEEN CELL-IMMEDIATED AND HUMORAL IMMUNITY

CELL-IMMEDIATED HUMORAL IMMUNITY
IMMUNITY
Immune response mediated by cells Immune response mediated by antibodies
Protects against fungi, viruses, and Protects against extracellular bacterial pathogens
facultative intracellular bacterial and viruses infecting respiratory or intestinal tract;
pathogens and prevents recurrence of viral infections
Mediates delayed (type IV) Mediates immediate (types I, II, and III)
hypersensitivity hypersensitivity
Only T-cell-dependent antigens lead to B cells directly bind soluble antigens resulting in
cellmediated immunity production of antibodies
Both CD4+ and CD8+ T cells are involved Only TH cells are involved
Provides immunological surveillance and No major role in immunological surveillance
immunity against cancer
Participates in rejection of homografts and May be involved in early graft rejection due to
graftversus-host reaction preformed antibodies

Key Points
Adaptive immunity shows the following features:
■ It is the resistance acquired by an individual during life.
■ It occurs after exposure to an agent and is mediated by antibodies as well as T lymphocytes (helper
T cells and cytotoxic T cells).
■ It has immunologic memory and a remarkable capability of discriminating between self and nonself
antigens.
■ Once an antigen has been recognized by the cells of acquired immune system, the response to it is
specific and can be repeated. In most cases, the acquired immune response improves with repeated
exposure.
■ The immune response to the second challenge occurs more quickly than the first, is stronger, and is
often more effective in neutralizing and clearing the pathogen
DIFFERENCES BETWEEN PASSIVE AND ACTIVE IMMUNITY
PASSIVE IMMUNITY ACTIVE IMMUNITY
No active host participation; received Produced actively by host’s immune system
passively
Antibodies transferred directly Antibodies induced by infection or by immunogens
Passive immunity is due to readymade Active immunity often involves both the cell-
antibodies mediated and humoral immunity
Types: Natural—transfer of maternal Types: Natural—clinical or inapparent infection;
antibodies through placenta; Artificial— Artificial— induced by vaccines
injection of immunoglobulins
Immediate immunity; no lag period Immunity effective only after lag period
Transient; less effective Durable; effective protection
No immunological memory Immunological memory present
Subsequent dose less effective due to Booster effect on subsequent dose
immune elimination
No negative phase Negative phase may occur
Applicable even in immunodeficient Not applicable in immunodeficient

■ HUMORAL IMMUNITY: It is mediated by molecules in the blood and mucosal secretions

called antibodies. The antibodies are secreted by a subset of lymphocytes known as B cells. The
antibodies recognize microbial antigens, combine specifically with the antigens, neutralize the
infectivity of microbes, and target microbes for elimination by various effector mechanisms. Humoral
immunity is the principal defense mechanism against extracellular microbes.
■ CELL-MEDIATED IMMUNITY: It is mediated by both activated TH cells and CTLs. Cytokines
secreted by TH cells activate various phagocytic cells, enabling them to phagocytose and kill
microorganisms. This type of cell-mediated immune response is especially important against a host of
bacterial and protozoal pathogens. CTLs play an important role in killing virus-infected cells and
tumor cells. They act by killing altered self-cells.
ANTIGEN RECOGNITION: Antigens, which are generally very large and complex, are not
recognized in their entirety by lymphocytes. Instead, both B and T lymphocytes recognize discrete
sites on the antigens called antigenic determinants, or epitopes. Epitopes are the immunologically
active regions on a complex antigen, the regions that actually bind to B-cell or T-cell receptors.
B cells and T cells differ in their mechanisms of antigen recognition. While B cells recognize the
antigen by interacting with the epitope on their own, T cells recognize the antigen only when the
epitope is “presented” by one of the specialized antigen-presenting cells. Once the antigen has been
recognized, these cells then go on to diversify by several intricate mechanisms. This diversification
helps in conferring the specificity, one of the cardinal characteristics of the immune system.
MAJOR HISTOCOMPATIBILITY COMPLEX (MHC): It is a large genetic complex with
multiple loci. The MHC loci encode two major classes of membrane-bound glycoproteins: class I and
class II MHC molecules. Class II molecules present antigens to the TH cells, while class I molecules
do the same for CTLs. In order for a foreign protein antigen to be recognized by a T cell, it must be
degraded into small antigenic peptides that form complexes with class I or class II MHC molecules.
This conversion of proteins into MHC-associated peptide fragments is called antigen processing and
presentation.
PASSIVE IMMUNITY
When immunity is conferred by transfer of serum or lymphocytes from a specifically immunized
individual, it is known as passive immunity. This is a useful method for conferring resistance rapidly,
i.e., without waiting for the development of an active immune response. Passive immunity may be
natural or artificial.
NATURAL PASSIVE IMMUNITY: It is observed when IgG is passed from mother to fetus during
pregnancy. This forms the basis of prevention of neonatal tetanus in neonates by active immunization
of pregnant mothers. It is achieved by administering tetanus toxoid to pregnant mothers during the
last trimester of pregnancy. This induces production of high level of antibodies in mother against
tetanus toxin, which are subsequently transmitted from mother to fetus through placenta. The
antibodies subsequently protect neonates after birth against the risk of tetanus. Natural passive
immunity is also observed by passage of IgA from mother to newborn during breast feeding.
ARTIFICIAL PASSIVE IMMUNITY: It is induced in an individual by administration of
preformed antibodies, generally in the form of antiserum, raised against an infecting agent.
Administration of these antisera makes large amounts of antibodies available in the recipient host to
neutralize the action of toxins. The preformed antibodies against rabies and hepatitis A and B viruses,
etc. given during incubation period prevent replication of virus, and hence alter the course of
infection. Immediate availability of large amount of antibodies is the main advantage of passive
immunity. However, short life span of these antibodies and the possibility of hypersensitivity
reaction, if antibodies prepared in other animal species are given to individuals who are
hypersensitive to these animal globulins (e.g., serum sickness), are the two noted disadvantages of
passive immunity
Combined passive–active immunity is carried out by giving both preformed antibodies (antiserum)
and a vaccine to provide immediate protection and long-term protection, respectively, against a
disease. This approach is followed for prevention of certain infectious conditions, namely, tetanus,
rabies, and hepatitis B.
LOCAL IMMUNITY The immunity at a particular site, generally at the site of invasion and
multiplication of a pathogen, is referred to as local immunity. Local immunity is conferred by
secretory IgA antibodies in various body secretions. These antibodies are produced locally by plasma
cells present on mucosal surfaces or in secretory glands. Natural infection or attenuated live viral
vaccines given orally or intranasally induces local immunity at gut mucosa and nasal mucosa,
respectively.
HERD IMMUNITY Herd immunity refers to an overall level of immunity in a community.
Eradication of an infectious disease depends on the development of a high level of herd immunity
against the pathogen. Epidemic of a disease is likely to occur when herd immunity against that
disease is very low indicating the presence of a larger number of susceptible people in the
community.

GENERAL CONCEPTS

A. Specific and Nonspecific Defenses

The protection of our organism against infectious agents involves many different mechanisms—some
nonspecific (i.e., generically applicable to many different pathogenic organisms) and others specific
(i.e., their protective effect is directed to one single organism).
Nonspecific defenses, which as a rule are innate (i.e., all normal individuals are born with them),
include:
              Mechanical barriers such as the integrity of the epidermis and mucosal membranes
hysicochemical barriers, such as the acidity of the stomach fluid
              The antibacterial substances (e.g., lysozyme, defensins) present in external secretions
Normal intestinal transit and normal flow of bronchial secretions and urine, which eliminate
infectious agents from the respective systems
              Ingestion and elimination of bacteria and particulate matter by granulocytes, which is
independent of the immune response
Specific defenses, as a rule, are induced during the life of the individual as part of the complex
sequence of events designated as the immune response. The immune response has two unique
characteristics:
a.           Specificity for the eliciting antigen; for example, immunization with inactivated poliovirus
only protects against poliomyelitis, not against viral influenza. The specificity of the immune
response is due to the existence of exquisitely discriminative antigen receptors on lymphocytes. Only
a single or a very limited number of similar structures can be accomodated by the receptors of any
given lymphocyte. When those receptors are occupied, an activating signal is delivered to the
lymphocytes. Therefore, only those lymphocytes with specific receptors for the antigen in question
will be activated. 
b.           Memory, meaning that repeated exposure to a given antigen elicits progressively more
intense specific responses. Most immunizations involve repeated administration of the immunizing
compound, with the goal of establishing a long-lasting, protective response. The increase in the
magnitude and duration of the immune response with repeated exposure to the same antigen is due to
the proliferation of antigen-specific lymphocytes after each exposure. The numbers of responding
cells will remain increased even after the immune response sub-sides. Therefore, whenever the
organism is exposed again to that particular anti-gen, there is an expanded population of specific
lymphocytes available for activation, and, as a consequence, the time needed to mount a response is
shorter and the magnitude of the response is higher.

B. Stages of the Immune Response

To better understand how the immune response is generated, it is useful to consider it as divided into
separate sequential stages .The first stage, induction, involves a small lymphocyte population with
specific receptors able to recognize an antigen or antigen fragments generated by specialized cells
known as antigen-presenting cells (APCs). 

The proliferation and differentiation of APCs is usually enhanced by amplification systems involving
the APCs themselves and specialized T-cell subpopulations (T helper cells, defined below). This is
followed by the production of effector molecules (antibodies) or by the differentiation of effector
cells (cells that directly or indirectly mediate the elimination of un-desirable elements). The final
outcome, therefore, is the elimination of the organism or compound that triggered the reaction by
means of activated immune cells or by reactions triggered by mediators released by the immune
system.
ANTIGEN
Molecules that can be recognized by the immunoglobulin receptor of B cells or by the T-cell receptor
when complexed with major histocompatibility complex (MHC) are called antigens. The word
antigen is a shortened form of the words “antibody generator.” Antigens are substances that react
with antibodies, while immunogens are molecules that induce an immune response. In most cases,
antigens are immunogens, and the terms are used interchangeably. The antigens that are not
immunogenic but can take part in immune reactions are termed as haptens. The term
immunogenicity means the ability of an antigen to elicit an immune reaction in the form of a B-cell
or T-cell response, whereas the term antigenicity means just the ability to combine specifically with
the products of the above responses. All molecules that are immunogenic are antigenic too, but all
antigenic molecules cannot be considered immunogenic. Thus, haptens can be said to lack
immunogenicity. Determinants of Antigenicity A number of factors have been identified that make a
substance immunogenic.
IMPORTANT DETERMINANTS OF ANTIGENICITY
1. Molecular size
2. Foreignness
3. Chemical-structural complexity
4. Stability
5. Other factors
Molecular Size In general, protein molecules with large molecular weight are highly antigenic.
Substances with molecular weights of about 100,000 Da and more are highly immunogenic, while
substances with molecular weights of Da less than 5000 are generally not immunogenic. This
property has been exploited in experimental studies by using high molecular weight proteins like
bovine gamma globulin (MW 150,000 Da) to induce an immune reaction. Substances with low
molecular weight may be made antigenic by adsorbing these on carrier particles, such as bentonite,
kaolin, and other inert particles.
Foreignness To be immunogenic, a molecule must be recognized as nonself, i.e., foreign. The
molecule is considered self or nonself by the immune system depending on whether or not the
molecule was exposed to the immune system during fetal development. Foreignness implies ability of
the host to tolerate selfantigens. Tolerance to self-antigens develops by contact with them in the initial
phases of the development of immune system, particularly during the development of lymphocytes.
In general, the more distantly related two species are, the greater the immunogenicity of a molecule
from one species will be when exposed to the other. For example, the bovine serum albumin is more
immunogenic in a chicken than in a goat. A graft from an unrelated human will be rejected within
about 2 weeks unless immunosuppressive drugs are used, but a graft from a chimpanzee will be
rejected within hours even if drugs are used. In contrast, a kidney graft from an identical twin will be
accepted readily.
Chemical-Structural Complexity Proteins are the most potent immunogens followed by
polysaccharides. Nucleic acids and lipids are not efficient in eliciting a good immune reaction,
although they may act as haptens. Structural complexity of a protein contributes to its
immunogenicity. Chains of single amino acids or single sugars are poorly immunogenic, but if
different amino acids or sugars are combined in the same molecule, the immunogenicity is greatly
enhanced. In cell-mediated immunity, the response of T cells to the peptide component of the proteins
depends on how the peptide is recognized and presented by the MHC cells. Therefore, the structure of
protein plays an important role in its immunogenicity, especially in inducing cellular immunity. The
lipid-specific antibodies are not easily produced; hence, they do not play a major role in immunity.
However, these antibodies have a role in the measurement of certain lipid-based molecules and drugs.
These antibodies are produced first by treating lipids with haptens and then conjugating with suitable
carrier molecules, such as the proteins (e.g., hemocyanin or bovine serum albumin).
Stability Highly stable and nondegradable substances (e.g., some plastics, metals, or chains of D-
amino acids) are not immunogenic. This is because internalization, processing, and presentation by
antigen-presenting cells (APCs) are always essential to mount an immune response. Therefore, very
stable substances (such as silicon) have been successful as nonimmunogenic materials for
reconstructive surgeries, such as breast implants. On the other hand, if a substance is very unstable, it
may break up before an APC can be internalized, and hence become immunogenic. In addition, large,
insoluble complexes are more immunogenic than smaller, soluble ones. This is because macrophages
find it easier to phagocytose, degrade, and present the insoluble complexes than the soluble
complexes.
Other Factors
◗ Biological system
Biological system also plays an important role in determining the immunological efficiency of an
antigen. Some substances are immunogenic in one individual but not in others (i.e., responders and
nonresponders). This is due to the fact that individuals may lack or have altered genes that code for
the receptors for antigen on B cells and T cells, or they may not have the appropriate genes needed for
the APC to present antigen to the helper T (TH) cells.
◗ Dosage and route of the antigen
The dose of antigen and the route by which it comes into contact with the immune system also
influence immunogenicity of the antigen. Very low doses of antigen do not stimulate immune
response, either because too few lymphocytes are contacted or because a nonresponsive state is
elicited. Conversely, an extremely high dose also fails to elicit tolerance. Repeated administration of
antigens (booster doses) may be required to enhance immune response of the host to certain antigens.
This is particularly important in case of vaccines where a prerequisite immune level needs to be
attained. Hence the booster doses of vaccines, such as DPT (Diphtheria, Pertussis, Tetanus), DT
(Diphtheria, Tetanus), etc., are given to ensure good protective levels of antibodies. Generally,
antigens are administered by the parenteral route to produce good level of antibodies. The antigens
can be given by (a) intravenous, (b) subcutaneous, (c) intradermal, (d) intramuscular, (e)
intraperitoneal, and (f) mucosal routes. Usually, the subcutaneous route of administration proves to be
better than intravenous routes at eliciting an immune response.
◗ ADJUVANTS
Adjuvants are the substances that when mixed with an antigen and injected with it boost the
immunogenicity of the antigen. Adjuvants increase both the strength and the duration of immune
response. Adjuvants boost immunogenicity of antigens in several ways:
■ Adjuvants like aluminum potassium sulfate (alum) and Freund’s water-in-oil adjuvant prolong the
persistence of antigen by forming a depot at the injection site. Alum precipitates the antigen and
releases it a little at a time. The water-in-oil emulsion forms small droplets with the antigen and also
releases these slowly over time.
■ Freund’s complete adjuvant contains, in addition to the emulsifying factors, heat-killed
mycobacteria. The bacterial components activate macrophages and increase both the production of
IL-1 and the level of B7 membrane molecules, which enhances the immune response. The increased
expression of class II MHC increases the ability of APC to present antigen to TH cells. B7 molecules
on the APC bind to CD28, a cell-surface protein on TH cells, triggering costimulation, an
enhancement of the T-cell immune response.
■ Some adjuvants, like synthetic polyribonucleotides and bacterial lipopolysaccharides, stimulate
nonspecific lymphocyte proliferation and bring about their action.

ANTIGENIC SPECIFICITY
Antigenic specificity of the antigen depends on antigenic determinants or epitopes.
EPITOPES
An epitope is defined as the immunologically active region of an immunogen that binds to antigen-
specific membrane receptors on lymphocytes or secreted antibodies. The interaction between cells of
the immune system and antigens takes place at many levels and the complexity of any antigen is
mirrored by its epitope. There are two types of epitopes: B-cell epitopes and T-cell epitopes.
◗ B-CELL EPITOPES
B-cell epitopes are antigenic determinants recognized by B cells. B-cell epitope can combine with its
receptor only if the antigen molecule is in its native state. The complementary surfaces of the
antibody and the antigen molecules appear to be relatively flat. Smaller molecules often fit nicely
within a particular depression or groove in the antigen-binding site of the antibody molecule. The B-
cell epitope is about six or seven sugar residues or amino acids long. B-cell epitopes tend to be
hydrophilic and are often located at bends in the protein structure. They are also often found in
regions of proteins, which have a higher mobility; this may make it possible for an epitope to shift
just a bit to fit into an almost-right site.
◗ T-CELL EPITOPES
T cells recognize amino acids in proteins but do not recognize polysaccharide or nucleic acid
antigens. This is the reason why polysaccharides are considered as T-independent antigens and
proteins as T-dependent antigens. The primary sequence of amino acids in proteins determines the
antigenic determinants recognized by T cells. Free peptides are not recognized by T cells, while the
complex of MHC molecules and peptide are recognized by T cells. Thus for a T-cell response, it
should recognize both the antigenic determinant and also the MHC, and therefore it is said to be
MHC restricted. In general, T-cell epitopes or antigenic determinants are small and are only 8–15
amino acids long. The antigenic determinants are limited to those parts of the antigen that can bind to
MHC molecules. Since the MHC molecules are subjected to genetic variability, there can be
difference among individuals in their T-cell response to the same stimulus. Each MHC molecule can
bind several, but not all, peptides. Therefore, for a peptide to be immunogenic in a particular
individual, that individual must have MHC molecules that can bind to it.
KEY POINTS
Processing of an antigen by APCs is absolutely essential for a T cell to recognize it. Two different
types of processing can prepare a protein antigen for antigen presentation. These include:
■ Externally derived antigens’ processing: In this process, phagocytosed bacteria are killed and lysed
by phagocytic cells, such as macrophages. Pieces of the bacteria are then processed and presented in
the context of class II MHC molecules.
■ Endogenously derived antigens’ processing: In this process, virus proteins synthesized in a cell are
processed and then presented in the context of class I MHC molecules.
SPECIES SPECIFICITY Tissues of all individuals in a species possess certain speciesspecific
antigens. However, some degree of cross-reaction occurs between antigens from related species. The
species specificity shows phylogenetic relationship. The phylogenetic relationship is useful in:
■ Tracing evolutionary relationship between species.
■ The species identification from blood and seminal stains in forensic medicine.
ISOSPECIFICITY
Isospecificity is determined by the presence of isoantigens or histocompatibility antigens.
ISOANTIGENS Isoantigens are antigens found in some, but not all, members of a species. A species
may be grouped depending on the presence of different isoantigens in its members. These are
genetically determined. Human erythrocyte antigens, based on which individuals are classified into
different blood groups, are the best examples of isoantigens in humans. The blood groups are of
primary importance in:
■ Transfusion of blood and blood products,
■ Isoimmunization during pregnancy, and
■ Providing valuable evidence in paternity disputes, the results of which are supplemented by more
recent DNA fingerprinting tests.
HISTOCOMPATIBILITY ANTIGENS
Histocompatibility antigens are the cellular determinants specific for each individual of a species.
These antigens are associated with the plasma membrane of tissue cells. Human leukocyte antigen
(HLA) is the major histocompatibility antigen that determines the homograft rejection. Therefore,
HLA typing is absolutely essential before carrying out transplantation of tissue or organ from one
individual to another.
AUTOSPECIFICITY
Self-antigens are generally nonantigenic. Sequestrated antigens (such as eye lens protein and sperm)
are, however, exceptions, because these are not recognized as selfantigens. This is because corneal
tissue and sperm are never encountered by the immune system during the development of tolerance to
self-antigens. Therefore, these tissues become immunogenic if accidentally or experimentally
released into the blood or tissues.
ORGAN SPECIFICITY
Antigens characteristic of an organ or tissues are called organ-specific antigens. These antigens found
in the brain, kidney, and lens tissues, even of different animal species, share the same antigen
specificity. Organ-specific antigens, such as brain-specific antigens, shared by human and sheep brain
are one such example. The antirabies vaccines prepared from sheep brain, when given, may induce
immune response in some humans, causing damage to neural tissues of the recipient. This may result
in neuroparalytic complications in some individuals.
HETEROPHILE SPECIFICITY
Heterophile specificity is determined by the presence of heterophile antigens. The same or closely
related antigens, sometimes present in tissues of different biological species, classes, or kingdoms are
known as heterophile antigens. Antibodies against the heterophile antigens produced by one of the
species cross-react with the antigens of other species. This property is exploited for diagnosis of
many infectious diseases. Weil–Felix reaction, Paul-Bunnell test, and cold agglutination tests are the
examples of serological tests that use such heterophile antigens.
KEY POINTS
■ Weil–Felix reaction is a test used for diagnosis of rickettsial infections, in which the strains of
Proteus species (such as OX 19, OX 2, and OX K) are used to detect heterophile antibodies produced
against rickettsial pathogens.
■ Paul–Bunnell test is used for diagnosis of infectious mononucleosis caused by Epstein–Barr virus
infection by demonstration of heterophile antibodies that agglutinate sheep erythrocytes.
■ Cold agglutinin test is performed for diagnosis of primary atypical pneumonia caused by
Mycoplasma pneumoniae by demonstration of heterophilic antibodies.
HAPTENS
Haptens are small organic molecules that are antigenic but not immunogenic. They are not
immunogenic because they cannot activate helper T cells. Failure of hapten to activate helper T cells
is due to their inability to bind to MHC proteins; they cannot bind because they are not proteins and
only proteins can be presented by MHC proteins. Moreover, haptens are univalent hence cannot
activate B cells by themselves. The haptens, however, can activate B cells when covalently bound to
a “carrier” protein. When bound with a carrier molecule, they form an immunogenic hapten–carrier
conjugate. In this process, the haptens combine with an IgM receptor on the B cells, and the hapten–
carrier protein complex is internalized. A peptide of the carrier protein is presented in association
with class II MHC protein to the helper T cells. The activated helper T cells then produce
interleukins, which stimulate the B cells to produce antibodies to hapten. Animals immunized with
such a conjugate produce antibodies specific for (a) the hapten determinant, (b) unaltered epitopes on
the carrier protein, and (c) new epitopes formed by combined parts of both the hapten and carrier. In
fact, the hapten–carrier molecule is bound to surface immunoglobulins on B cells via the hapten
epitopes. The hapten–carrier molecule is then taken in, processed, and pieces of the carrier are
presented by these B cells and TH cells. In the body, the formation of hapten–carrier conjugates is the
basis for development of allergic responses to drugs, such as penicillin.
SUPERANTIGENS
Superantigens are a class of molecules that can interact with APCs and T lymphocytes in a
nonspecific way. The superantigens act differently by interacting with MHC class II molecules of the
APC and the Vb domain of the T-lymphocyte receptor. This interaction results in the activation of a
larger number of T cells (10%) than conventional antigens (1%), leading to massive cytokine
expression and immunomodulation. Examples of superantigens are staphylococcal enterotoxins, toxic
shock syndrome toxin, exfoliative toxins, and also some viral proteins.

ANTIBODIES
Antibodies are globulin proteins (immunoglobulins) that are synthesized in serum and tissue fluids,
which react specifically with the antigen that stimulated their production. Three types of globulins are
present in the blood: alpha, beta, and gamma. The antibodies are the gamma globulins. Antibodies are
one of the major plasma proteins, and against infection often referred to as “first line of defense”. The
most important function of antibodies is to confer protection against microbial pathogens.
Antibodies confer protection in the following ways:
1. They prevent attachment of microbes to mucosal surfaces of the host.
2. They reduce virulence of microbes by neutralizing toxins and viruses.
3. They facilitate phagocytosis by opsonization of microbes.
4. They activate complement, leading to complement-mediated activities against microbes. Von
Behring and Kitasato performed the first experiments that proved the physical existence of antibodies
in 1890. They demonstrated that serum obtained from rabbits immunized with tetanus or diphtheria
toxins could prevent disease in mice infected with such pathogens. The unknown substance that was
present in serum and that provided protection on transfer was named “antitoxin” by Tizzoni and
Cattani in 1891. Subsequently, experimental works by Paul Ehrlich and Jules Bordet demonstrated
that a protective response could be generated even against whole cells (erythrocytes). The more
inclusive term antibody subsequently replaced the term antitoxin. Tiselius and Kabat accomplished
the first successful attempt to identify antibody molecules in 1939. They demonstrated that
hyperimmunization increased the concentration of globulins in serum and that this fraction contained
antibody activity. Because globulins are large-molecular-weight proteins, it was suggested that
further characterization of antibodies requires breaking them into smaller and easily handled
fragments. Porter in 1959, succeeded in digesting rabbit immunoglobulin G (IgG) with the proteolytic
enzyme papain. These produced two distinct fragments: a monovalent fragment with antigenbinding
activity, termed Fab (fragment antigen binding) and a second fragment that retained the antibody’s
effector functions and crystallized readily into a lattice, termed Fc (fragment crystallizable).
Edelman and Poulik using a similar method splitted myeloma globulins into two distinct
components, which subsequently were termed heavy (H) and light (L) chains. The World Health
Organization (WHO) in 1964 coined the term “immunoglobulin (Ig)” for the term antibody. The
immunoglobulin includes not only antibody globulins but also the cryoglobulins, macroglobulins, and
abnormal myeloma proteins. Thus, all antibodies are immunoglobulins but not all immunoglobulins
may be antibodies.
IMMUNOGLOBULINS
There are five classes of immunoglobulins: (i) immunoglobulin G (IgG), (ii) immunoglobulin M
(IgM), (iii) immunoglobulin A (IgA), (iv) immunoglobulin E (IgE), and (v) immunoglobulin D (IgD).
Myeloma proteins were first used for the amino acid sequencing of immunoglobulins. These proteins
were also the first immunoglobulins that were subjected to crystallographic studies. They provided
the first glimpses of the domain structure of the prototypic immunoglobulin. Structure of
Immunoglobulins Immunoglobulins show the following properties: ■ They are glycoproteins. ■ They
are a complex structure of four polypeptide chains: two identical heavy (typically 55 kDa each)
chains and two identical light chains (25 kDa each). This gives immunoglobulin an overall ‘Y’ or ‘T’
shape, which is the most widely recognized feature of immunoglobulin structure. ● The terms
“heavy” and “light” refer to the molecular weights of the chains. The heavy chains have a molecular
weight of 50,000–70,000 Da, while light chains have a molecular weight of 25,000 Da. The heavy
chains are longer, and light chains are shorter
◗ Heavy chains
An immunoglobulin molecule has two heavy chains. Each heavy chain is made up of 420–440 amino
acids. The two heavy chains are held together by one to five disulfide (S—S) bonds. Each heavy
chain is bound to a light chain by a disulfide bond and by noncovalent bonds, such as salt linkages,
hydrogen bonds, and hydrophobic bonds to form a heterodimer (H–L). Similar noncovalent
interactions and disulfide bridges link the two identical heavy and light (H–L) chains to each other to
form the basic four-chain (H–L)2 antibody structure. The heavy chains of a given antibody molecule
determine the class of that antibody. For example, IgM contains mu (µ), IgG contains gamma (¥),
IgA contains alpha (α), IgD contains delta (), and IgE contains epsilon (£). These heavy chains are
structurally and antigenically distinct for each class of immunoglobulin. They differ in their size,
carbohydrate content, and as antigens.
◗ LIGHT CHAINS An immunoglobulin molecule has two light chains. Each light chain is made up
of 220–240 amino acids. Light chain is attached to the heavy chain by a disulfide bond. The light
chains are structurally and chemically similar in all classes of immunoglobulins. They are of two
types: kappa (k) and lambda (ƛ). These two types differ in their amino acids present in constant
regions. Each immunoglobulin has either two k or two ƛ chains but never both. The k and ƛ chains
are present in human serum in a ratio of 2:1.
◗ VARIABLE AND CONSTANT REGIONS
Each polypeptide chain of an immunoglobulin molecule contains an amino terminal part and a
carboxy terminal part. The amino terminal part is called the variable region (V region) and the
carboxy terminal part is called the constant region (C region). Both heavy and light chains contain
variable and constant regions. These regions are composed of three-dimensional folded structures
with repeating segments, which are called domains. Each heavy chain consists of one variable (VH)
and three constant (CH) domains. IgG and IgA have three CH domains (CH1, CH2, and CH3),
whereas IgM and IgE have four domains (CH1, CH2, CH3, and CH4). Each light chain consists of
one variable (VL) and one constant domain (CL).
VARIABLE REGION: The amino-terminal half of the light or heavy chain, consisting of 100–110
amino acids, is known as variable or V regions (VL in light chains and VH in heavy chains). V region
is different for each class of immunoglobulin. The variable regions of both light and heavy chains
consist of three highly variable regions known as hypervariable regions. The antigen combining sites
Fab of the antibody molecule that consists of only 5–10 amino acids each are present in the
hypervariable region of both the light and heavy chains. These antigen-binding sites are responsible
for specific binding of antibodies with antigens. The high specificity of antibodies is primarily due to
the presence of these hypervariable regions.
CONSTANT REGION:
The carboxyl-terminal half of the molecule is called the constant (C) region. It consists of two basic
amino acid sequences. The Fc fragment, found to crystallize under low ionic conditions, is present in
the constant region of heavy chain. The constant region of the heavy chain has many biological
functions. It is responsible for activation of the complement, binding to cell surface receptors,
placental transfer, and many other biological activities. The constant region of the light chain has no
biological function. A single antibody molecule has two identical heavy chains and two identical light
chains, H2L2, or a multiple (H2L2) n of this basic four-chain structure. Subisotypes exist for and
chains, and this leads to the existence of subclasses of the respective immunoglobulins.
IMMUNOGLOBULIN ANTIGEN DETERMINANTS
There are three major types of immunoglobulin antigen determinants: isotypes, allotypes, and
idiotypes.
◗ ISOTYPES
The isotype of an immunoglobulin refers to the particular constant region of the light- or heavy-chain
of the immunoglobulin. Immunoglobulins are classified on the basis of various heavy chain isotypes.
Heavy chains are distinguished by the presence of heavy chain markers, such as µ, gamma , α, delta,
€, in the immunoglobulins IgM, IgG, IgA, IgD, and IgE, respectively. The light chains are also
distinguished by isotype markers, such as k and ƛ. Isotypes are present in all members of a species.
◗ ALLOTYPES
The allotype refers to allelic differences in both the variable and constant regions of immunoglobulin.
The allotype markers are present on the constant regions of light and heavy chains. They are Am on α
heavy chains, Gm on gammaheavy chains, and Km on k light chains. Allotype markers are absent on
µ, gamma, and epsilon heavy chains and on lambda light chains. More than 25 Gm types, 3 Km
allotypes, and 2 Am on IgA have been described. Allotypes are present in some but not all members
of a species and are inherited in a simple Mendelian fashion.
◗ IDIOTYPES
The idiotype refers to a specificity that is associated with the variable region. Idiotype markers are
found on the hypervariable region of the immunoglobulin. Idiotypes are specific for each antibody
molecule. Anti-idiotypic antibodies produced against Fab fragments prevent antigen–antibody
interaction.

BIOSYNTHESIS OF IMMUNOGLOBULINS
B lymphocytes and plasma cells take part in the synthesis of immunoglobulins. Resting B cells
synthesize only small amounts of immunoglobulins that mainly get incorporated into cell membranes.
Plasma cells, the most differentiated B cells, are specialized to produce and secrete large amounts of
immunoglobulins. The synthetic capacity of the plasma cells is reflected by the abundant cytoplasm,
which is extremely rich in endoplasmic reticulum. Normally, heavy and light chains are synthesized
in separate polyribosomes of the plasma cell. The amounts of heavy and light chains synthesized on
the polyribosomes are usually balanced and so both types of chains are combined to produce
complete Ig molecules, without excess of any given chain. The assembly of a complete Ig molecule is
carried out either by associating one heavy and one light chain to form an H–L hemi-molecule, and
then joining two H–L hemi-molecules to form a single complete molecule (H2L2), or by forming H2
and L2 dimers that later associate to form the complete molecule. While free light chains can be
effectively secreted from plasma cells, free heavy chains are generally not secreted. The heavy chains
are synthesized and transported to the endoplasmic reticulum, where they are glycosylated, but
secretion requires combination with light chains to form a complete immunoglobulin molecule. If
light chains are not synthesized or heavy chains are synthesized in excess, the free heavy chains
combine through their CH1 domain with a heavy-chainbinding protein, which is believed to be
responsible for their intracytoplasmic retention. Both IgM and IgA are the polymeric antibodies,
which have one additional polypeptide chain, the J chain. The J chain is synthesized by all plasma
cells, including those that produce IgG. However, it is only incorporated to polymeric forms of IgM
and IgA. It is believed that the J chain has some role in initiating polymerization. IgM proteins are
assembled in two steps. First, the monomeric units are assembled. Then, five monomers and one J
chain combine via covalent bonds to produce a pentameric molecule.

IMMUNOGLOBULIN CLASSES
The structure and biological functions of five classes of immunoglobulins (IgG, IgM, IgA, IgE, and
IgD) are described below:
◗ Immunoglobulin G
IgG is a 7S immunoglobulin with a molecular weight of 150,000 Da. It has a half-life of 23 days—
longest among all the immunoglobulins. IgG is the most abundant class of immunoglobulins in the
serum, comprising about 80% of the total serum immunoglobulin. There are four IgG subclasses
IgG1, IgG2, IgG3, and IgG4 so numbered according to their decreasing concentrations in serum.
Though the differences between these subclasses are minute, their functions vary as follows:
1. IgG1, IgG3, and IgG4 are special because these are the only immunoglobulins with the ability to
cross the placental barrier. They play an important role in protecting the developing fetus against
infections.
2. IgG3, IgG1, and IgG2, in order of their efficiency, are effective in the activation of the
complement.
3 IgG1 and IgG3 bind with high affinity to Fc receptors on phagocytic cells and thus mediate
opsonization. IgG4 has an intermediate affinity for Fc receptors and IgG2 has an extremely low
affinity. Two gammachains, along with two k or gamma light chains, joined together by disulfide
bonds, comprise an IgG molecule as follows:
■ The gamma chain is a 51-kDa, 450-amino acid residue heavy polypeptide chain.
■ It consists of one variable VH domain and a constant (C) region with three domains designated
CH1, CH2, and CH3.
■ The hinge region is situated between CH1 and CH2.
■ Proteolytic enzymes, such as papain and pepsin, cleave an IgG molecule in the hinge region to
produce Fab and F (ab´) 2 and Fc fragments. There are four subclasses of IgG in humans with four
corresponding chain isotypes designated gamma-1, -2, -3, and -4. IgG1, IgG2, IgG3, and IgG4 show
differences in their hinge regions and differ in the number and position of disulfide bonds that link
two gamma chains in each IgG molecule. There is only a 5% difference in amino acid sequence
among human gamma chain isotypes, exclusive of the hinge region. Cysteine residues, which make it
possible for interheavy (gamma) chain disulfide bonds to form are found in the hinge area. IgG1 and
IgG4 have two interheavy chain disulfide bonds, IgG2 has 4, and IgG3 has 11. The IgG, is distributed
equally in the intra- and extravascular compartments.
Key Points
IgG shows the following biological activities:
■ In response to infection, IgG antibodies appear late after appearance of IgM antibodies, but persists
for a longer period.
■ It confers protection against the microorganisms that are present in the blood and tissues. It is
distributed equally in the intra- and extravascular compartments.
■ It is the only immunoglobulin that crosses the placenta; hence, it confers natural passive immunity
to the newborns.
■ It takes part in precipitation, complement fixation, and neutralization of toxins and viruses.
■ It binds to microorganisms and facilitates the process of phagocytosis of microorganisms.

IMMUNOGLOBULIN M
IgM constitutes about 5–8% of total serum immunoglobulins. It is distributed mainly intravascularly.
It is a heavy molecule (19S) with a molecular weight varying from 900,000 to 1,000,000 Da
(millionaire molecule). It has a half-life of 5 days. IgM is basically a pentamer, composed of five
immunoglobulin subunits (monomeric subunits, IgMs) and one molecule of J chain. Each monomeric
IgM is composed of two light chains (k or gamma light chains) and two heavy chains (µ). The heavy
chains are larger than those of IgG by about 20,000 Da, corresponding to an extra domain on the
constant region (CH4). Two subclasses of IgM (IgM1 and IgM2) are described, which differ in their
µ chains. IgM1 consists of µ1 and IgM2 consists of µ2 chains . The immunoglobulin µ chain is a 72
kDa, 570-amino acid heavy polypeptide chain comprising one variable region, designated VH, and a
four-domain constant region, designated CH1, CH2, CH3, and CH4. The µ chain does not have a
hinge region. A “tail piece” is located at the carboxy terminal end of the chain. It comprises 18-amino
acid residues. A cysteine residue at the penultimate position of a carboxy terminal region of the µ
chain forms a disulfide bond that joins to the J chain. There are five N-linked oligosaccharides in the
µ chain of humans. Monomeric IgM, with a molecular weight of 180,000 Da, is expressed as
membrane-bound antibody on B cells. As mentioned earlier, the J chain found in the IgM molecule
was believed to play a major role in the secretion of its polymerized form. Being present on the
membrane of B cells, IgM acts as the antigen-binding molecule in the antigen–antibody complex.
Because of its pentameric structure with 10 antigenbinding sites, serum IgM has a higher valency
than the other isotypes. An IgM molecule can bind 10 small hapten molecules; however, because of
steric hindrance, only five or fewer molecules of larger antigens can be bound simultaneously.
Treatment of serum with 2-mercaptoethanol destroys IgM without affecting IgG antibodies. This
forms the basis for differential estimation of IgM and IgG antibodies in serum pretreated with 2-
mercaptoethanol

KEY POINTS
IgM shows the following biological activities:
■ Pentameric IgM, because of its high valency, is more efficient than other isotypes in binding
antigens with many repeating epitopes, such as viral particles and red blood cells.
■ It is more efficient than IgG in activating complement. Complement activation requires two Fc
regions in close proximity, and the pentameric structure of a single molecule of IgM fulfills this
requirement.
■ IgM is the first immunoglobulin produced in a primary response to an antigen. The
immunoglobulin confers protection against invasion of blood by microbial pathogens. Deficiency of
IgM antibodies is associated with septicemia.
■ IgM antibodies are short lived and disappear early as compared to IgG. The presence of IgM
antibody in serum, therefore, indicates recent infection.
■ It is also the first immunoglobulin to be synthesized by a neonate in about 20 weeks of age. IgM is
not transported across the placenta; hence, the presence of IgM in the fetus or newborn indicates
intrauterine infection. The detection of IgM antibodies in serum, therefore, is useful for the diagnosis
of congenital infections, such as syphilis, rubella, toxoplasmosis.
◗ IMMUNOGLOBULIN A
IgA is the second major serum immunoglobulin, comprising nearly 10–15% of serum
immunoglobulin. It has a half-life of 6–8 day . IgA consist of α heavy chain that confers class
specificity on IgA molecules. The α chain is a 58-kDa, 470-amino acid residue heavy polypeptide
chain. The chain is divisible into three constant domains, designated CH1, CH2, and CH3, and one
variable domain, designated VH. Hinge region is situated between CH1 and CH2 domains. An
additional segment of 18-amino acid residues at the penultimate position of the chain contains a
cysteine residue where the J chain can be attached through a disulfide bond. IgA occurs in two forms:
serum IgA and secretory IgA.
Serum IgA: It is present in the serum and is a monomeric 7S molecule with a molecular weight of
60,000 Da. It has a half life of 6–8 days. It has two subclasses, IgA1 and IgA2, which are two α-chain
isotypes α-1 and α-2, respectively. The α-2 chain has two allotypes, A2m (1) and A2m (2), and does
not have disulfide bonds linking heavy to light chains. Differences in the two α chains are found in
two CH1 and five CH3 positions. Thus, there are three varieties of α-heavy chains in humans.
SECRETORY IGA: It is a dimer or tetramer and consists of a J-chain polypeptide and a polypeptide
chain called secretory component, or SC, or secretory piece .The SC is a polypeptide with a
molecular weight of 70,000 Da and is produced by epithelial cells of mucous membranes. It consists
of five immunoglobulin-like domains that bind to the Fc region domains of the IgA dimer. This
interaction is stabilized by a disulfide bond between the fifth domain of the SC and one of the chains
of the dimeric IgA. IgA-secreting plasma cells are concentrated along mucous membrane surfaces.
The daily production of secretory IgA is greater than that of any other immunoglobulin. Secretory
IgA is the major immunoglobulin present in external secretions, such as breast milk, saliva, tears, and
mucus of the bronchial, genitourinary, and digestive tracts. IgA activates the complement not by
classical pathway but by alternative pathway
KEY POINTS
Secretory IgA shows the following biological activities:
■ It protects the mucous membranes against microbial pathogens. It serves an important effector
function at mucous membrane surfaces, which are the main entry sites for most pathogenic
organisms. Because it is polymeric, secretory IgA can cross-link large antigens with multiple
epitopes.
■ Binding of secretory IgA to bacterial and viral surface antigens prevents attachment of the
pathogens to the mucosal cells, thus inhibiting viral infection and bacterial colonization. Complexes
of secretory IgA and antigen are easily entrapped in mucus and then eliminated by the ciliated
epithelial cells of the respiratory tract or by peristalsis of the gut.
■ Breast milk contains secretory IgA and many other molecules that protect the newborns against
infection during the first month of life. Because the immune system of infants is not fully functional,
breast-feeding plays an important role in maintaining the health of newborns.
■ Secretory IgA has shown to provide an important line of defense against bacteria (such as
Salmonella spp., Vibrio cholerae, and Neisseria gonorrhoeae) and viruses (such as polio, influenza,
and reovirus).
KEY POINTS
Secretory IgA shows the following biological activities:
■ It protects the mucous membranes against microbial pathogens. It serves an important effector
function at mucous membrane surfaces, which are the main entry sites for most pathogenic
organisms. Because it is polymeric, secretory IgA can cross-link large antigens with multiple
epitopes.
■ Binding of secretory IgA to bacterial and viral surface antigens prevents attachment of the
pathogens to the mucosal cells, thus inhibiting viral infection and bacterial colonization. Complexes
of secretory IgA and antigen are easily entrapped in mucus and then eliminated by the ciliated
epithelial cells of the respiratory tract or by peristalsis of the gut.
■ Breast milk contains secretory IgA and many other molecules that protect the newborns against
infection during the first month of life. Because the immune system of infants is not fully functional,
breast-feeding plays an important role in maintaining the health of newborns.
■ Secretory IgA has shown to provide an important line of defense against bacteria (such as
Salmonella spp., Vibrio cholerae, and Neisseria gonorrhoeae) and viruses (such as polio, influenza,
and reovirus).
◗ IMMUNOGLOBULIN E
IgE constitutes less than 1% of the total immunoglobulin pool. It is present in serum in a very low
concentration (0.3 ug/mL). It is mostly found extravascularly in lining of the respiratory and intestinal
tracts. IgE is an 8S molecule with a molecular weight of 190,000 Da and half-life of 2–3 days. Unlike
other immunoglobulins that are heat stable, IgE is a heat-labile protein—easily inactivated at 56°C in
1 hour . Two e heavy polypeptide chains, along with two k or two ƛ light chains, fastened together by
disulfide bonds, comprise an IgE molecule. The e chain is a 72-kDa, 550-amino acid residue
polypeptide chain. It consists of one variable region, designated VH, and a four-domain constant
region, designated CH1, CH2, CH3, and CH4. This heavy chain does not possess a hinge region. In
humans, the epsilon heavy chain has 428 amino acid residues in the constant region. IgE does not
cross the placenta or fix the complement.
KEY POINTS
IgE shows the following biological activities:
■ IgE is also known as reaginic antibody that mediates the type I immediate hypersensitivity (atopy)
reactions.
■ IgE is responsible for the symptoms of hay fever, asthma, and anaphylactic shock. IgE binds to Fc
receptors on the membranes of blood basophils and tissue mast cells. Cross-linkage of receptor bound
IgE molecules by antigen (allergen) induces basophils and mast cells to translocate their granules to
the plasma membrane and release their contents to the extracellular environment—a process known
as degranulation. As a result, varieties of pharmacologically active mediators are released and give
rise to allergic manifestations.
■ Localized mast-cell degranulation induced by IgE may also release mediators that facilitate a
buildup of various cells necessary for antiparasitic defense
◗ IMMUNOGLOBULIN D
IgD comprises less than 1% of serum immunoglobulins. It is a 7S monomer with a molecular weight
of 180,000 Da. The halflife of IgD is only 2–3 days. IgD has the basic fourchain monomeric
structure with two delta heavy chains (molecular weight 63,000 Da each) and either two k or two ƛ
light chains (molecular weight 22,000 Da each) . Immunoglobulin delta chain is a 64-kDa, 500-amino
acid residue heavy polypeptide chain consisting of one variable region, designated as VH, and a
three-domain constant region, designated as CH1, CH2, and CH3. There is also a 58-residue amino
acid residue hinge region in human delta chains. Two exons encode the hinge region. IgD is very
susceptible to the action of proteolytic enzymes at its hinge region. Two separate exons encode the
membrane component of delta chain. A distinct exon encodes the carboxy terminal portion of the
human delta chain that is secreted. The human delta chain contains three N-linked oligosaccharides.
KEY POINTS
IgD is present on the surface of B lymphocytes and both IgD and IgM serve as recognition receptors
for antigens. The role of IgD in immunity continues to remain elusive.
ABNORMAL IMMUNOGLOBULINS
Abnormal immunoglobulins are other structurally similar proteins that are found in serum in certain
pathological conditions, such as multiple myeloma, heavy chain disease, and cryoglobulinemia and
sometimes in healthy individuals also.
Multiple myeloma: Bence-Jones (BJ) proteins were the earliest abnormal proteins described in 1847
that were found in patients with multiple myeloma. These proteins are the light chains of
immunoglobulins, hence occur as either k or ƛ forms. In a patient, it may occur as either k or ƛ but
never in both the forms. BJ proteins have a peculiar property of coagulating at 60°C and redissolving
again at a higher temperature of 80°C. In multiple myeloma, plasma cells synthesizing IgG, IgA, IgD,
or IgE are affected. Myeloma involving IgM-producing plasma cells is known as Waldenström’s
macroglobulinemia. This condition is characterized by excessive production of the respective
myeloma proteins (M proteins) and that of their light chains (BJ proteins). The study of myeloma
proteins led to a great advancement in our understanding of immunoglobulin function. These “single”
or “monoclonal” antibodies obtained from the sera of patients with multiple myeloma were used in
many of the serologic and biochemical studies of the 1950s and 1960s. They remained the major
source of homogeneous immunoglobulins until the development of the hybridoma in 1974. The
serologists injected them into animals and produced antisera that were used to study some of the basic
properties of antibodies. For example, the immune sera were absorbed with other myeloma proteins
and were used to identify isotypic, allotypic, and idiotypic specificities.
HEAVY CHAIN DISEASE: Heavy chain disease is a different disorder, which is a lymphoid
neoplasia, characterized by an excess production of heavy chains of the immunoglobulins.
CRYOGLOBULINEMIA: Cryoglobulinemia is a condition characterized by presence of
cryoglobulins in blood. The condition may not be always associated with disease but is often found in
patients with macroglobulinemia, systemic lupus erythematosus, and myelomas. Most cryoglobulins
consist of either IgG or IgM or their mixed precipitates. In cryoglobulinemia, serum from patient
precipitates on cooling and redissolves on warming