BMS2.

2
Immunochemistry
 Basic Concept of Immunochemistry

• The immune system is a remarkable defense

mechanism which provides the means to make rapid,
specific, and protective responses against the myriad
of potentially pathogenic organisms that inhabit the
world in which we live.
• The immune system comprises a range of cells,
tissues and chemicals that interact to overcome
infection, repair tissue damage and maintain the
integrity of the body
 Causes of Disease

• Pathogenic Organisms
• Genetic Disorders
• Toxic Chemicals
• Other Environmental Factors
• Physical Damage to Organs
• Nutritional Disorders
Types of Pathogenic Organisms
• Viruses
• Bacteria
• Protozoans
• Fungi
• Animals
• Parasites
 Mechanisms of
Disease by Pathogens
• Utilization of host nutritional
resources
• Physical damage to host tissues
• Production of toxic substances
• Chromosomal and gene damage
• Body cells behave abnormally
Self vs. Non-self: How does the body know?

• Self refers to particles, such as proteins and

other molecules, that are a part of, or made
by your body.
• Non-self refers to particles that are not
made by your body, and are recognized as
potentially harmful. These are sometimes
called foreign bodies. Non-self-particles or
bodies can be bacteria, viruses, parasites,
pollen, dust, and toxic chemicals.
 Antigens
• Antigens are anything that causes an immune
response. Antigens can be entire pathogens,
like bacteria, viruses, fungi, and parasites, or
smaller proteins that pathogens express.
 Cytokines
• Cytokines are molecules that are used for cell
signaling, or cell-to-cell communication.
Cytokines are can be used to communicate
with neighboring or distant cells about
initiating an immune response. Cytokines are
also used to trigger cell trafficking, or
movement, to a specific area of the body.
 Chemokines
• Chemokines are a type of cytokines that are
released by infected cells. Infected host cells
release chemokines in order to initiate an
immune response, and to warn neighboring
cells of the threat.
 Innate and Acquired immune system

• The immune response has been described

as comprising the non-specific or innate
response and the acquired or specific
response.
• The innate response is a generalized
response irrespective of the precipitating
agent.
• By contrast, the specific immune response
involves the precise recognition of particles
that are foreign to the host’s body (i.e. they
are not normally present in the healthy
body and are called non-self).
 Types of immune response

Pathogens and their disease causing by-products are eradicated

by three mechanisms.

Nonspecific Immunity—physical and chemical barriers

including the action of phagocytic cells

Antibody Mediated Immunity (Humoral Immunity)—free

circulating antibodies found in the blood and lymph fluids
that are effective against viruses, bacteria and toxins

Cell Mediated Immunity—leads to killing by cells through

the recognition of antigens that are present on pathogen
infected cells
Defense Mechanisms
1. External defense
2. Internal Defense
3. Immune Defense
First Line of Defense
 Innate Immune system
Second line of defense

• 1. Cells of the immune system are generally

derived from pluripotent hematopoietic
stem cells in the bone marrow.
• There are many types of white blood cells, or
leukocytes, that work to defend and protect
the human body.
• In order to patrol the entire body, leukocytes
travel by way of the circulatory system.
•
1. Cells of the Immune System
 Phagocytes or Phagocytic cells
• Phagocyte means “eating cell”, which describes what role
phagocytes play in the immune response. Phagocytes
circulate throughout the body, looking for potential threats,
like bacteria and viruses, to engulf and destroy. You can think
of phagocytes as security guards on patrol.
 Macrophages
• Macrophages are efficient phagocytic cells that can
leave the circulatory system by moving across the
walls of capillary vessels.
• The ability to roam outside of the circulatory system
is important, because it allows macrophages to hunt
pathogens with less limits.
• Macrophages can also release cytokines in order to
signal and recruit other cells to an area with
pathogens.
 Mast cells
• Mast cells are found in mucous membranes and connective
tissues, and are important for wound healing and defense
against pathogens via the inflammatory response.
• When mast cells are activated, they release cytokines and
granules that contain chemical molecules to create an
inflammatory cascade.
• Mediators, such as histamine, cause blood vessels to dilate,
increasing blood flow and cell trafficking to the area of
infection.
• The cytokines released during this process act as a
messenger service, alerting other immune cells,
like neutrophils and macrophages, to make their
way to the area of infection, or to be on alert for
circulating threats.
 Neutrophils
• Neutrophils are phagocytic cells that are also
classified as granulocytes because they contain
granules in their cytoplasm.
• These granules are very toxic to bacteria and
fungi, and cause them to stop proliferating or die
on contact.
• The bone marrow of an average healthy adult
makes approximately 100 billion new neutrophils
per day.
• Neutrophils are typically the first cells to arrive at
the site of an infection because there are so many
of them in circulation at any given time.
 Eosinophils
• Eosinophils are granulocytes that target
multicellular parasites.
• Eosinophils secrete a range of highly toxic
proteins and free radicals that kill bacteria and
parasites.
• The use of toxic proteins and free radicals also
causes tissue damage during allergic reactions, so
activation and toxin release by eosinophils is
highly regulated to prevent any unnecessary
tissue damage.
 Basophils
• Basophils are also granulocytes that attack
multicellular parasites.
• Basophils release histamine, much like mast
cells.
• The use of histamine makes basophils and
mast cells key players in mounting an allergic
response.
 Natural Killer cells
• Natural Killer cells (NK cells), do not attack pathogens
directly.
• Instead, natural killer cells destroy infected host cells
in order to stop the spread of an infection.
• Infected or compromised host cells can signal natural
killer cells for destruction through the expression of
specific receptors and antigen presentation.
 Dendritic cells
• Dendritic cells are antigen-presenting cells that are
located in tissues, and can contact external
environments through the skin, the inner mucosal
lining of the nose, lungs, stomach, and intestines.
• Since dendritic cells are located in tissues that are
common points for initial infection, they can
identify threats and act as messengers for the rest
of the immune system by antigen presentation.
• Dendritic cells also act as bridge between the
innate immune system and the adaptive immune
system.
 General Immune Response
• General Immune Responses - such as non-specific
cellular responses, inflammation, Fever and
complement.
• The inflammatory response actively brings
immune cells to the site of an infection by
increasing blood flow to the area.
 Inflammation
Non-specific reaction to stimuli such as toxins or pathogens.

Mediated by a subgroup of leukocytes (white blood cells) that produce

cytokines that lead to fibrin clots at the site of inflammation.

The inflammatory response results in redness, swelling, heat and pain at

the site of the infection

Most important outcome is the immobilization of the pathogen at the

site of inflammation
 Fever

Normal body temperature is 37 degrees Celsius

Abnormal increase in the body temperature—usually caused by

infectious agents

Certain products of pathogenic bacteria can be pyrogenic—

EXAMPLE:
Endotoxin (LPS) from gram-negative bacteria.
Some bacteria cause the release of endogenous pyrogens from the
WBCs that kill them.

Slight temperature increases boost the immune system by increasing

The activity of phagocytic cells and antibody responses to pathogens.

Strong fevers (40 degrees Celsius) harm the host by damaging

host tissues
 Complement system

• The complement system (also called the

complement cascade) is a mechanism that
complements other aspects of the immune
response.
• Typically, the complement system acts as a part of
the innate immune system, but it can work with
the adaptive immune system if necessary.
• complement is a complex network of plasma and
membrane-associated serum proteins which can
elicit highly efficient and tightly regulated
inflammatory and cytolytic immune responses to
• infectious organisms(bacteria, viruses, parasites),
• tissue damaged by physical, chemical, or
neoplastic insults,
• and other surfaces identified as ‘non-self’
• Activation of complement leads to robust
and efficient proteolytic cascades, which
terminate in opsonization and lysis of the
pathogen as well as in the generation of the
classical inflammatory response through
the production of potent pro-inflammatory
molecules.
 The complement system
• The complement system refers to a series of >20 proteins,
circulating in the blood and tissue fluids.
• Most of the proteins are normally inactive, but in response to
the recognition of molecular components of microorganisms
they become sequentially activated in an enzyme cascade.
• The activation of one protein enzymatically cleaves and
activates the next protein in the cascade.
• Complement can be activated via three different pathways
which can each cause the activation of C3, cleaving it into
a large fragment, C3b, that acts as an opsonin, and a
small fragment C3a (anaphylatoxin) that promotes
inflammation.
• Activated C3 can trigger the lytic pathway, which can
damage the plasma membranes of cells and some
bacteria.
• C5a, produced by this process, attracts macrophages and
neutrophils and also activates mast cells.
 Classical Pathway

• This pathway involves complement components C1, C2

and C4. The pathway is triggered by antibody-antigen
complexes binding to C1, which itself has three
subcomponents C1q, C1r and C1s.
• The pathway forms a C3 convertase, which splits C3 into
two fragments; the large fragment, C3b, can covalently
attach to the surface of microbial pathogens and
opsonise them.
• The small fragment, C3a, activates mast cells, causing the
release of vasoactive mediators such as histamine.
 Alternative Pathway

• This pathway involves various factors, B, D, H & I, which

interact with each other, and with C3b, to form a C3
convertase, that can activate more C3, hence the pathway is
sometimes called ‘the amplification loop’.
• Activation of the loop is promoted in the presence of bacterial
and fungal cell walls, but is inhibited by molecules on the
surface of normal mammalian cells.
 Mannose-binding Lectin Pathway

• This pathway is activated by the binding of

mannose-binding lectin (MBL) to mannose
residues on the pathogen surface.
• This in turn activates the MBL-associated serine
proteases, MASP-1 and MASP-2, which activate
C4 and C2, to form the C3 convertase.
 Lytic Pathway

• This pathway is initiated by the splitting of C5, and

attachment of C5b to a target. C6, C7, C8 and C9 unite with
C5b, and this membrane-attack complex (MAC).
• when inserted into the outer membrane of some bacteria,
can contribute to their death by lysis.
Acquired, Adaptive or Specific immune system

• The acquired, adaptive or specific immune response

involves the precise recognition of particles that are
foreign to the host’s body (i.e. they are not normally
present in the healthy body and are called non-self).
• These may be molecules on host cells, which have been
altered in some way, or invading microorganisms.
• The cells involved in mediating these responses are found
in the blood and in specialized tissues throughout the
body (the lymphoid tissues).
Lymphoid Tissues
• Adaptive immunity creates immunological
memory after an initial response to a specific
pathogen, and leads to an enhanced response to
subsequent encounters with that pathogen.
• This process of acquired immunity is the basis of
vaccination.
• Like the innate system, the adaptive system
includes both humoral immunity components and
cell-mediated immunity components.
• Adaptive immunity can also provide long-lasting
protection; for example, someone who recovers
from measles is now protected against measles
for their lifetime.
• In other cases it does not provide lifetime
protection; for example, chickenpox.
• The adaptive system response destroys invading
pathogens and any toxic molecules they produce.
• Sometimes the adaptive system is unable to
distinguish harmful from harmless foreign
molecules; the effects of this may be hay fever,
asthma or any other allergy.
• The cells that carry out the adaptive immune
response are white blood cells known as
lymphocytes.
• Two main broad classes: antibody responses (B-
Cells) and cell mediated immune response (T
cells).
• In antibody responses, B cells are activated to
secrete antibodies, which are proteins also known
as immunoglobulins.
• The adaptive immune response is triggered by
recognizing foreign antigen in the cellular context
of an activated dendritic cell.
 Antigen Presentation
• With the exception of non-nucleated cells (including
erythrocytes), all cells are capable of presenting antigen
through the function of major histocompatibility complex
(MHC) molecules.
• Some cells are specially equipped to present antigen, and to
prime naive T cells.
• Dendritic cells, B-cells, and macrophages are equipped with
special "co-stimulatory" ligands recognized by co-stimulatory
receptors on T cells, and are termed professional antigen-
presenting cells (APCs).
• With the exception of non-nucleated cells (including
erythrocytes), all cells are capable of presenting antigen
through the function of major histocompatibility complex
(MHC) molecules.
• Some cells are specially equipped to present antigen, and
to prime naive T cells.
Major Histocompatibility complex proteins are found
on the surface of cells:: T cells cannot recognize foreign antigens
unless they are associated with these MHC proteins

Class I MHC proteins are Class II MHC proteins are only

found on the surface of ALL found on the surface of
nucleated cells B lymphocytes, macrophages
and other antigen presenting cells

ALL MHC proteins are imbedded in the cytoplasmic membrane of

cells and project outward from the cell surface
• Several T cells subgroups can be activated by professional
APCs, and each type of T cell is specially equipped to deal
with each unique toxin or microbial pathogen.
• The type of T cell activated, and the type of response
generated, depends, in part, on the context in which the
APC first encountered the antigen