INFECTION AND IMMUNITY

Immunity is the resistance exhibited by the host towards injury caused by micro-
organisms and their products. Protection against the infectious diseases is only one
arm of the immune response, since the main purpose is the reaction of the body
against foreign antigen.

Immune cells and the responses they generate. There are 2 types of immunity:

1. INNATE - this is also called the NATURAL IMMUNITY since it is present even at
birth. This response will not increase upon repeated exposure to the antigen.
It is composed of skin, mucous membranes, secretions (saliva, tears),
phagocytic cells and NK cells. Innate immunity also allows elimination of
foreign substance like a bacteria, without previous exposure. It is enhanced
by natural antibodies or natural cytotoxic cells like macrophages, neutrophils,
eosinophils and NK cells. It is nonspecific since it is effective against a wide
range of infectious agents.

Factors which affect innate immunity: Age (infants are affected more by
infections since they have an immature immune system); Sex(death rate is
higher in males, but there is no marked differences in susceptibility between
males and females);Hormonal influence (there is decreased resistance to
infection in DM);Nutrition (poor nutrition leads to increased susceptibility to
infections).

MECHANISMS OF INNATE IMMUNITY

A. MECHANICAL BARRIERS AND SURFACE SECRETIONS

The intact skin and mucous membranes of the body give avery good
protecton against pathogens. The skin is a resistant barrier because of
its outer cornified layer consisting mostly of keratin which micro-
organisms cannot digest. Also, the dry condition of the skin plus the
high concentration of salt in the sweat is lethal.

Sebaceous secretions and sweat also contain bactericidal and

fungicidal fatty acids.

The sticky mucus of the respiratory tract acts as a trapping mechanism

for inhaled pathogens. Nasal secretions and saliva also contain
mucoploysaccharides which can block some viruses.

The washing of tears and flushing of urine are effective in invasion or

organisms.
 There are also normal bacterial flora in the epithelial surfaces which
are beneficial: they occupy the space which cannot be occupied
anymore by pathogens, they have used up nutrients; they produce by-
rproducts which are harmful to pathogens.

B. HUMORAL DEFENSE MECHANISMS

A number of microbicidal substancesare present in the tissues and body

fluids.

Lysozyme is a protein found in high concentrations in neutrophils and in body

fluids, except for the CSF, sweat and urine. It functions as a mucolytic
enzyme against gram (+) cel wall, causing their lysis.it may calso cause the
intracellular destruction of gram (-) bacteria.

Basic polypeptides like spermine and spermidine can kill tubercle bacilli and
staphylococci.

Acute-phase proteins rise highly during an infection.Endotoxin can cause the

release of Interleukin-1, which in turn, stimulates the liver to produce the
acute-phase proteins. The most commonly known is C-reactive protein(CRP).
This bins to the bacterial cell wall. It also activates the classical complement
pathway. Alpha 1-antitrypsin, alpha 2- macroglobulin, fibrinogen and amyloid
are examples.

Interferon are factors released by cells in response to a viral infection. It

protected other cells of the same species from being infected by that virus.

C. COMPLEMENT

These are about 30 proteins found in the serum in their inactive forms, but
can be activated to form an enzyme cascade with the following end result:

Opsonization

Cellular activation

Cell lysis

There are two pathways: the classical and the alternative pathway

The classical pathway is the main antibody-directed mechanism for

complement activation; it is rapid and most efficient. The complement
recognizes the antigen-antibody complexes (either IgM or IgG), then bind to
C1, which then activates the cascade.
 The alternative pthway protects the body from pathogens in the absence of
antibody; it is activated by the bacterial surface itself.

The final result for both pathways is the formation of the membrane attack
complex, which is formed from the reactions among C5, C6, C7, C8 and C9.

D.CELLS

Phagocytes are cells which engulf micro-organisms as they enter tissues, fluids or
the bloodstream. Mononuclear phagocytes in the blood are called monocytes,
macrophages in tissues, histiocytes in connective tissue, mesangial cells in the
kidney, osteoclasts in bone, microglia in the brain, and sinus-lining macrophages in
the spleen, lymph node and thymus.

Phagocytes are:

Actively phagocytic

Contain digestive enzymes to degrade ingested material

Process and present antigens

Produce molecules that stimulate lymphocyte differentiation into effector cells.

Phagocytosis involves recognition, binding, ingestion and digestion of the cell.

Products of injured tissues, blood factors, leukotrienes and histamine from mast
cells and neutrophils, bacterial products

Chemotaxis

Phagocytes have receptors for the Fc portion of immunoglobulins and some

components of the complement cascade; these act as OPSONINS on the bacterial
cell surface and enhances the digestion process

The foreign particle is engulf by the cell membrane and internalized as an ENDSOME
or PHAGOSOME

LYSOSOMES in the phagocyte fuse with the phagosome forming the

PHAGOLYSOSOME
 Enzymes are released, killing the bacterial cell:superoxide dismutase and
myeloperoxidase, lyzozyme and elastase, lactoferrin

NATURAL KILLER CELLS recognize changes in virus-infected cells and destroy them
by an extracellular mechanism. This is present even without prior exposure to the
infectious agent. It is performed by cell large granular lymphocytes, and by cells
with T cell markers. Its activity is enhanced by Interferon. They have also been
implicated in the host defense against cancer cells.

EOSINOPHILS are polymorphonuclear cells with a blobed nucleus and cytoplasmic

granules. They are present in very low levels in the blood, but their numbers
increase with parasitic infections and allergies. They are not efficient phagocytic
cells, but their granules contain molecule which are toxic to microorganisms. These
released molecules target large parasites such as worms.

E.TEMPERATURE

Pyrexia which follows many infections can also be a source of control. It is mediated
by interleukin-1, which is produced by macrophages. It is also known as the
endogenous pyrogen.

F.INFLAMMATION

This is the reaction of the body to injury, such as invasion by a infectious agent,
exposure to a toxix chemical, or physical trauma. The five signs are redness, heat,
swelling, pain and loss of function.

The molecular events that happen in inflammation are:

Vasodilatation

Increased vascular permeability

Cellular infiltration.

The most common mediators of infection are:

Mediator Main source Function

Histamine mast cells vasodilatation, increased vascular

permeability

Basophils smooth muscle contraction

Kinins plasma same as above, plus pain

Prostaglandins neutrophils vasodilatation, increased permeability, pain

Eosinophils

Monocytes

Platelets

Leukotrienes neutrophils same as histamine, plus induce cell

adherence and

Chemotaxis

Complement cascade plasma cause the mast cells to release

C5 which is for chemotaxis

Plasmin plasma breaks down fibrin, kinin formation

Cytokines lymphocytes chemotactic factors, colony-stimulating

factors

Macrophages macrophage activation

ACQUIRED IMMUNITY develops late in fetal life, and development continues into
childhood. Continued exposure to the antigen stimulates acquired immunity. The
response is specific to an individual antigen because of the antigen-specific
recognition by surface antibody on B cells and by T-cell receptors on T cells.

Acquired immunity may have an anamnestic response, in which subsequent

response to a previously recognized antigen is more magnified due to the
production of memory T and B cells.

There are two types of acquired immunity: active and passive. Active immunity is
induced after contact with foreign antigen, such as micro organisms and their
products. This contact may be subclinical or clinical infection.

Immunization with live or killed infectious agents, exposure to microbial products

and transplantation of foreign cells is a form of active immunity. Long term
resistance , although slow in onset, occurs in acquired immunity.
Passive immunity is the prompt availability of large amounts of antibodies, and is
transmitted by antibodies or lymphocytes which have be preformed in another
host. There is however, a short life span,and the possibility of hypersensitivity
reactions since it is formed in another species. An example is the transfer of
maternal antibodies to the fetus.

An ANTIGEN is a substance that is capable of provoking the lymphoid tissues of an

animal to respond by generating an immune reaction. This reaction is specifically
directed against the inducing substance only and not at any other related
substances.

The response is just to some parts of the antigen, or individual chemical groups, and
not to the entire antigen itself. This is termed the specificity to antigenic
determinants or to epitopes. Thus ,since a micro-organism may have several
epitopes, more than one antibody will react with that particular organism.

Functional classification of ANTIGENS

1. Immunogens, substances that are able to elicit an immune response by

themselves

2. Haptens, molecules that are able to react with antibodies, but are unable to
stimulate their production directly; low molecular weight; become active only
when attached to carriers, which are large molecules

A substance that act an as antigen in one species may not necessarily produce a
reaction in another species.

Antigens, in general, must have a molecular weight of more than 5000. However,
even lower molecular weight substances like glucagon , can function as an antigen
with they are combined with an adjuvant, which in turn, causes additional stimulus
to the immune response.

Aspirin, penicillin and formaldehyde may also act as antigens although they have
low molecular weight.

ANTIGENIC DETERMINANTS

A response to antigen involves the specific interaction of components of the

immune sysem, antibodies and lymphocytes, with epitopes on the antigen.

The lymphocytes have receptors on their surface that function as the recognition
unit:
 Surface-bound immunoglobulin on B cells

T cell receptor on T cells

The part of the lymphocyte that attaches to the epitopes is called a PARATOPE.

Antigenic determinants may be formed in two ways:

1. May be contained in a single segment of primary sequence(SEQUENTIAL

EPITOPES)

2. May be assembled from residues far apart in the sequence, but are
brought together in close proximity due to folding of the
molecule(CONFORMATIONAL EPITOPES)

ANTIGENIC SPECIFICITY is based on:

1. Subtle chemical differences between molecules

2. Foreignness of a substance to an animal can depend on the presence of

chemical groupings that are not normally found in the animal’s body.

The ability of the antibody to form a high-affinity interaction with an antigen

depends on the two molecules coming together in a very precise bond. The
antibody may bind to another, structurally similar antigen, but the bond will be
weaker.

CROSS REACTIVITY occurs when the antibody to a particular antigen is found

capable of combining with the with an apparently unrelated antigen.

For example, Forssman antigen is found in RBC and pneumococci and salmonella. It
is thus termed a HETEROPHILE ANTIGEN. The antibodies then clump the rbc, thus
they are called ISOHAEMAGGLUTININS.

IMMUNOGLOBULINS

19TH CENTURY, von Behring and Kitasato in Berlin discovered that the serum of an
appropriately immunized animals contained specific neutralizing substances. They
were called ANTIBODIES OR IMUNOGLOBULINS.

General characteristics of antibodies:

1. Glycoprotein
 2. Present in the serum and body fluids

3. Induced when immunogenic molecules are introduced into the host’s

lymphoid system

4. Reactive with, and are specific, to the antigen that induced their formation

Antiserum contains antibodies formed in reaction to the antigen. This is then

separated by electrophoresis into alpha, beta and gamma fractions. Most of the
immunoglobulins will be in the gamma fractions, hence they are also called
GAMMAGLOBULINS.

ANTIBODY STRUCTURE

All antibodies have the same basic structure: 2 light chains and 2 heavy chains.

The two light chains are one of two types: either a Kappa or a Lambda. The
molecular weight is about 25000. They are composed of 2 areas, which are called
DOMAINS, and these are approximately 110 amino acids. One end of the chain is
constant, and is identical in all members of the isotype. This is termed the CL
region.

The other end shows sequence variation, and is called the VL region.

The heavy chains determine the isotype, and have a molecular weight of 50000 to
70000. They also have two domains which show sequence variation ( VH); while the
other domain is similar for all isotypes (CH).

The tertiary structure of the VL and VH regions determines the shape of the antigen-
combining site or paratope.

The two chains are held together by disulfide bridges and non-covalent interactions.

Since the two light and two heavy chains are identical, there will be two binding
sites for the antigen (2 paratopes ) at the amino- terminal or N- terminal.

The carboxyl – terminal or C-terminal at the other end of the antibody will be the
same for all the members of the isotype. It is responsible for the biological
properties of the isotype.

There is an area in heavy chains which is called the HINGE REGION. Enzymes may
break up the antibody, dividing them into Fab and Fc region.

The Fab region (fragment antigen binding) contains the paratope.

The Fc (fragment crystallizable) is similar for all the antibodies of the same isotype.
The differences from the other isotypes define the biological activity of the antibody.
The variability in the amino acid sequence in the variable domains of the light and
heavy chains is not found over their entire length, but restricted to short segments
only. These segments show variation, thus are called HYPERVARIABLE REGIONS.
These are the areas that come in direct contact with the antigen.

5 CLASSES OR ISOTYPES

1. IgA

2. IgM

3. IgD

4. IgG

5. IgE

IgG

This is the major immunoglobulin of the serum, as well as the CSF and the lymph,
making up almost 75%. It has a molecular weight of 150,000 in humans. It is also
the major antibody of the secondary response, and is an important defense against
bacteria and viruses.

It is the only antibody to cross the placenta, so it is the highest antibody in the
newborn, protecting it in the first 4-6 months of life.

The levels may be abnormally raised in Malaria, Kala-azar and Myeloma.

Its functions are:

1. Activates the complement

2. Acts as an opsonin

3. Mediates antibody-dependent cellular immunity

IgA

This is the predominant antibody in seromucous secretions (saliva, tears,

colostrums, respiratory, GIT and GUT secretions).

Its functions are:

1. To specifically bind antigen at mucosal sites

 2. Protect mucous membranes from attack by bacteria and viruses.

IgM

This is the antibody produced during the immune response, especially to bacteria
and viruses. Due to its large size, this isotype is mainly confined to the
intravascular pool. The presence of IgM in the newborn may indicate intrauterine
infection such as Syphillis, Rubella, HIV infection and Toxoplasmosis.

Its functions are:

1. Most efficient in agglutination, complement fixation and other antigen-

antibody reactions

2. Can be produced in a fetus with an infection.

IgD

This account for less than 1% of circulating antibodies. It is very susceptible to

proteolytic attack , thus has a very short half-life in serum.

Its functions are:

1. B cell antigen receptor

2. Marker for mature B cells

3. Occurs on cells of some lymphatic leukemias

IgE

This is present in very low levels in the blood; found on the surface of mast cells and
basophils, which possess a receptor specific for the Fc part of the immunoglobulin.
It is also called the REAGINIC ANTIBODY since it is associated with allergic response
and immediate hypersensitivity reaction.

Functions:

1. It is associated primarily with allergic and parasitic infections.

ABNORMAL IMMUNOGLOBULINS

Bence Jones protein is found in multiple myeloma. It can be identified in the urine
by its characteristic property of coagulation when heated to 50C, but redissolving at
70C.

CRYOGLOBULINEMIA is a condition where a gel or precipitate forms on coolng the

serum, which redissolves on warming. This may not always be involved in diseases,
but it is often found in myeloma and SLE.

There are 3 terms to describe variants among antibody molecules.

1. Isotype refers to the subclass of the immunoglobulins ( G,A,M,E,D)

2. Allotype refers to the genetically determined difference in a molecule

between two members of the same species; this is also called genetic
polymorphism. It is due to the substitution of only one or two amino acids in
the constant regions of heavy and light chains. They have no biological
significance.

3. Idiotype refers to the unique, individual differences between antibodies of

different antigen binding specificities. They are determined by the structure
of the variable regions of antibodies.

MAJOR HISTOCOMPATIBILITY COMPLEX

This is a collection of highly polymorphic genes encoding the proteins that regulate
an immune response. These genes include the class I and class II cell surface
proteins, and the class III which encode for the complement proteins. In humans,
the MHC is called HUMAN LEUKOCYTE ANTIGENS (HLA). And are found on the short
arm of chromosome 6.

The HLA proteins are present on the cell surfaces that enable the T cells to
recognize and bind antigenic peptides(immune recognition).

HLA CLASS I ANTIGENS are membrane glycoproteins on the surface of ALL

nucleated cells and platelets. They are necessary for antigen recognition( on
viruses, intracellular bacteria, parasites, tumor antigens) by CD8+ cytotoxic T
lymphocytes. Pepetide of normal intracellular proteins are bound to HLA class I
proteins, and displayed on the cell surface. Proteins made by intracellular viruses,
bacteria, parasites or neoantigens made by tumor cells will also be displayed on
HLA class I proteins. In humans, the 3 types of class I genes are called HLA-B, HLA-
B, and HLA-C antigens.

HLA CLASS II ANTIGENS are proteins expressed on more restricted set of cells,
including the antigen-presenting cells (Langerhans cells, activated macrophages)B
cells, and thymic epithelial cells involved in T-cell maturation. They are necessary
for antigen recognition by Th cells. In humans, the class II genes include HLA-DR,
HLA-DQ and HLA-DP. Class II proteins are found on cells that interact directly with
the Th cells, including B cells, monocyte-macrophages, langerhans cells, dendritic
cells and thymic epithelium. The proteins made by extracellular bacteria or
parasites or injected vaccines will also be displayed on HLA class II proteins.

HLA CLASS III ANTIGENS. Class III genes encode for the complement components or
regulators of serum complement component complex.

Many diseases are associated with an increased frequency of the HLA antigens.
Ankylosing spondylitis (HLA-B27), rheumatoid arthritis (HLA-DR4), and IDDM (HLA-
DR3 and HLA-DR4).

Tissue typing for organ transplantation involves matching the HLA Class I and HLA
Class II antigens.