Good Morning!

Cellular
Immunity
 TOPICS INCLUDED
• Immunity: Introduction & Definition
• Types of immunity
• Antigens and Antigenic determinants
• Major Histocompatibility Complex(MHC)
• Antigen presentation
• Clonal selection
• Structural basis for receptor specificity
• Primary and Secondary immune responses
• Hematopoiesis and cytokines affecting
hematopoiesis
• Mature cells from myeloid progenitors
 Neutrophils
 Eosinophils
 Basophils and mast cells
 Monocytes and macrophages
• Mature cells from lymphoid precursors
 T cells
 B cells
 NK cells
• Oral Immune Responses
 In gingivitis
 In chronic periodontitis
• References
 Immunity

• Immunity is defined as the resistance of an

organism to infection, disease, or other
unwanted biological invasion.
• The central function of the immune system is
to protect the host against infection by
viruses, bacteria, fungi and other microbial
invaders.

• In addition, the immune apparatus carries out

an immune surveillance function by destroying
aberrant or malignant host cells.
• The immune system comprises both specific and
nonspecific components.

• Non-specific or innate immune responses include

an array of inflammatory reactions that are
operative regardless of whether the host has
been previously exposed to a given micro
organism.

• The magnitude of these responses is

approximately the same with each exposure.
• In contrast, specific or adaptive immune
responses are much more vigorous if the host
has had prior exposure to that microorganism.
 Types of Immunity

• The immune system responds to the

introduction of foreign molecules by the
induction of two main protective mechanisms:
• Humoral immunity is mediated by antibody
molecules, and cell mediated immunity
involves the direct action of immune cells.
 Humoral immunity
• Antibodies or immunoglobulins, are soluble protein
molecules present in blood, saliva and other body fluids
that can recognize and bind to the antigen that
stimulated their formation.

• Because of this interaction, a series of secondary

effector mechanisms , which serve ultimately to
eliminate the antigen from the body, are mobilized(eg.
After binding Ag, antibodies may neutralize viruses or
toxins, thus preventing their pathologic effects on host
tissue).
• In conjunction with complement(a series of
enzymes in plasma), antibodies mediate the
lysis and destruction of microorganisms.
• Antibodies also opsonize bacteria and other
particles , thus promoting their phagocytosis
and degradation by phagocytic cells such as
polymorphonuclear leukocytes(PMNs) and
macrophages.
• Antibodies are formed by a specialized
lymphocyte type, the bone marrow-derived or
B lymphocyte.
• After activation by antigen, B lymphoctes (or B
cells) proliferate and differenciate into
antibody-secreting plasma cells .
• Thus, although humoral immunity is mediated
by antibodies, antibodies are produced by
immune cells after their activation by antigen.
 Cellular immunity
• The other major category of immune
phenomena, cell-mediated immunity, depends
on the activation of a second lymphocyte type,
the thymus-derived T-lymphocyte or T cell.
• The reactions of cell mediated immunity
include delayed-type hypersensitivity reactions
(the prototype for which is the tuberculin skin
reaction).
• Delayed hypersensitivity reactions are
important in resistance to obligate
intracellular parasites, including many viruses,
fungi, and bacteria; most forms of graft
rejection and tumor immunity.
• These reactions depend on mechanisms that
do not involve antibody.
• Finally, cell mediated immunity also
encompasses many immunoregulatory
functions, including T cell up-regulation(help)
or down-regulation(supression) of antibody
production and other responses.
Antigens and Antigenic determinants
• An antigen or immunogen is defined as any
substance that after contact with the immune
system , elicits an immune response
constisting of antibody production , cell-
mediated immunity, or both.
• The only essential requirement is that the
antigen be structurally distinct from self
components(which otherwise, may lead to
autoimmune reactions).
• Many molecules can be antigenic, including
proteins, glycoproteins, lipoproteins, complex
carbohydrates and nucleic acids.
• Many compounds that have been synthesized
in the laboratory but do not exist in nature are
also antigenic.
• This illustrates the adaptive capacity of the
immune system to generate immune
responses to essentially any challenge.
• The basis for this adaptibility rests on an
evolved mechanism of generating many
different receptors for antigens, rather than
on a evolved repertoire of these receptors that
is common to all members of a species.
• In general, a positive relationship exists
between the molecular weight of an antigen
and the size of the immune response
generated.
• Structural complexity also plays a role, in that
proteins,which are made up of many different
amino acids are excellent antigens, whereas
carbohydrates, which consist of repeating
identical subunit structures , often are not.
• Antigenicity also involves a minimum size
requirement.
• For peptides, this corresponds to chains of
approximately 8 amino acids in length.
• Smaller peptides fail to elicit immune
responses unless they are linked to an
immunogenic entity such as a foreign protein,
the carrier (eg. Haptens such as dinitrophenol
linked to immunogenic carrier).
• The minimum size requirement for antigenicity
probably reflects the affinity of interaction of peptides
with Class II histocompatibility proteins on antigen
presenting cell (APC)eg. macrophages.

• Bacteria, viruses, and other particulate material such

as aggregated proteins tend to be highly immunogenic,
because they are readily engulfed by phagocytic cells,
which have an important antigen presentation function
in the induction of immune responses.
 Major Histocompatibility
Compex(MHC)
• The major histocompatibility complex (MHC) is
a large cluster of genes found on the short
arm of chromosome 6. The complex spans four
million base pairs of DNA and contains 128
genes as well as 96 pseudogenes (non-
functional gene remnants).
• Many, but not all of the genes in this complex
play important roles in the immune system.
• Traditionally, the MHC is divided into the class
I, II and III regions, each containing groups of
genes with related functions.
• The class I and II MHC genes encode human
leukocyte antigens (HLAs), proteins that are
displayed on the cell surface and define an
individual’s tissue type.

• There are many possible tissue types in the

population because each HLA exists as a large
number of varieties.
• Everyone's immune system is tolerant of its
own HLAs, but if foreign HLAs are detected
then the cells displaying them are attacked
and destroyed.
• This is why the body rejects grafts and
transplants from donors that have not been
matched for tissue type.
• The class I and II MHC proteins also perform
the important function of antigen
presentation.
• This is how the immune system finds out what
is happening inside the cells even though it
can only survey them from the outside.
• Proteins inside the cell are broken into short
fragments and displayed as peptide antigens
by MHC proteins on the surface.

• This helps the immune system to discriminate

between normal (self) antigens and those that
are foreign and potentially dangerous.
• Class I MHC proteins are found on virtually all
cell types and their job is to present fragments
of proteins that are synthesised inside the cell.
• The peptide antigens presented in this manner
are checked by killer T-cells, which have
receptors for the class I MHC proteins.
• The purpose of this surveillance system is to
identify abnormal body cells, such as those
infected with viruses or those that have turned
malignant.
• Such cells will display unfamiliar peptide
antigens, e.g. fragments of viral proteins, and
are attacked and destroyed.
• Class II MHC proteins are found only on
immune cells such as phagocytes that engulf
foreign particles such as bacteria.
• These cells are specially designed to present
peptide antigens derived from such digested
particles.
• The antigens are presented to helper T-cells,
which have receptors for class II MHC proteins.
• The purpose of this surveillance system is to
stop the immune system running out of
control and attacking the body's own cells.

• Only if the presented antigen is recognised as

foreign by the helper T-cell is the phagocyte
allowed to survive.
T cell receptor(TCR) – MHC binding
• Class III MHC genes encode several components
of the complement system, a collection of soluble
proteins found in the blood that targets foreign
cells and breaks open their membranes.
• Adjacent to the class III region is a group of genes
that control inflammation.
• Further genes with various immune and non-
immune functions are dotted throughout the
complex.
• The MHC shows a high degree of polymorphism
(100 times higher than the genome average, i.e.
a 10 per cent difference between any two
unrelated individuals).

• Many of these polymorphisms appear to be

associated with either increased or decreased
susceptibility to a range of infectious diseases
including malaria, tuberculosis, leprosy, typhoid
fever, hepatitis and HIV/AIDS.
• Defects in certain MHC genes lead to
autoimmune disorders in which the body fails
to recognise self-antigens.

• Examples of such diseases include multiple

sclerosis, some forms of arthritis and diabetes,
and inflammatory bowel disease.
 NAME FUNCTION EXPRESSION
MHC CLASS I Encodes heterodimeric peptide-binding All nucleated cells. MHC
proteins, as well as antigen-processing class I proteins contain an α
molecules such as TAP and Tapasin. chain & β2-micro-
globulin(not part of the
MHC). They present antigen
fragments to cytotoxic T-
cells and will bind to CD8 on
cytotoxic T-cells.
MHC CLASS II Encodes heterodimeric peptide-binding On antigen presenting cells,
proteins and proteins that modulate MHC class II proteins contain
antigen loading onto MHC class II α & β chains and they
proteins in the lysosomal compartment present antigen fragments
such as MHC II DM, MHC II DQ, MHC II to T-helper cells by binding
DR, and MHC II DP. to the CD4 receptor on the
T-helper cells.

MHC CLASS III Encodes for other immune components, Variable

such as complement components (e.g.,
C2, C4, factor B) and some that encode
cytokines (e.g., TNF-α).
 Antigen presentation
• Antigen presentation is a process in the
body's immune system by which
macrophages, dendritic cells and other cell
types capture antigens and then enable their
recognition by T-cells.
• The basis of adaptive immunity lies in the
capacity of immune cells to distinguish
between the body's own cells, and infectious
pathogens.
• The host’s cells express “self” antigens that
identify them as such.
• These antigens are different from those in
bacteria ("non-self" antigens) or in virally-
infected host cells (“missing-self”).
• The ability of the adaptive immune system to
survey for infection requires specialized
pathways of enabling recognition of
pathogen-derived antigens by T cells.
 Antigen recognition
• Unlike B cells, T cells fail to recognize antigen
in the absence of antigen-presentation, with
the important exception of the
superantigens.
• [Superantigens (SAgs) are secreted proteins
(exotoxins) that exhibit highly potent
lymphocyte-transforming (mitogenic) activity
directed towards T lymphocytes. Compared
to a normal antigen-induced T-cell response
where .001-.0001% of the body’s T-cells are
activated, SAgs are capable of activating up
to 20% of the body’s T-cells.
• This causes a massive immune response that is
not specific to any particular epitope on the
SAg. Since one of the fundamental strengths
of the adaptive immune system is its ability to
target antigens with high specificity, SAgs
produce an immune response that is
effectively useless. Microbes produce SAgs as a
defense mechanism to aid them in evading the
immune system.]
• The T cell receptor is restricted to recognizing
antigenic peptides only when bound to
appropriate molecules of the major
histocompatibility complex (MHC), also known
in humans as Human leukocyte antigen (HLA).
• Most cells are capable of presenting antigen
and activating the adaptive response.

• Some cells, however, are specially equipped to

acquire and present antigen, and to prime
naive T cells.
• Dendritic cells, B cells, and macrophages play
a major role in the innate response, and also
act as professional antigen presenting cells
(APC).
• These professional APCs are equipped with
special immunostimulatory receptors that
allow for enhanced activation of T cells.
• Several different types of T cells can be
activated by professional APCs, and each type
of T cell is specially equipped to deal with
different pathogens, whether the pathogen is
bacterial, viral or a toxin.
• The type of T cell activated, and therefore the
type of response generated, depends, in part,
on the context in which the antigen was first
encountered by the APC.
 Intracellular antigens: Class I
• Intracellular antigens are produced by viruses
replicating within a host cell.

• The host cell digests pathogen-associated

cytoplasmic proteins (in addition to
endogenous self-proteins, as well as tumor-
associated antigens) by a specialized enzyme
complex, the proteasome into small peptides.
• A specialized carrier, the Transporter
associated with Antigen Processing (TAP)
complex moves the peptide into the
endoplasmic reticulum, allowing the antigenic
peptide to be coupled to an MHC Class I
molecule and transported to the cell surface.
• MHC Class I molecules present antigen to
CD8+ cytotoxic T cells.

• With the exception of some cell types (such as

erythrocytes), Class I MHC is expressed by
almost all host cells.
• Cytotoxic T cells (also known as TC, killer T cell,
or cytotoxic T-lymphocyte (CTL)) are a
population of T cells which are specialized for
inducing the death of other cells.
• Recognition of antigenic peptides through
Class I by CTLs leads to the killing of the target
cell, which is infected by virus,
intracytoplasmic bacterium, or are otherwise
damaged or dysfunctional.
 Extracelluar antigens: Class II
• Dendritic cells (DCs) phagocytose exogenous
pathogens, such as bacteria, parasites or
toxins in the tissues and then migrate, via
chemotactic signals, to T cell enriched lymph
nodes.
• During migration, DCs undergo a process of
maturation in which they lose phagocytic
capacity and develop an increased ability to
communicate with T-cells in the lymph nodes.
• This maturation process is dependent on
signaling from other pathogen-associated
molecular pattern (PAMP) molecules through
pattern recognition receptors, such as the
members of the Toll-like receptor family.
• The DC uses lysosome-associated enzymes to
digest pathogen-associated proteins into
smaller peptides.
• In the lymph node, the DC will display these
antigenic peptides on its surface by coupling
them to MHC Class II molecules.
• This MHC:antigen complex is then recognized by
T cells passing through the lymph node.

• Exogenous antigens are usually displayed on

MHC Class II molecules, which interact with CD4 +
helper T cells. CD4+ lymphocytes, or TH , are
immune response mediators, and play an
important role in establishing and maximizing the
capabilities of the adaptive immune response.
• Expression of Class II is more restricted than
Class I.

• High levels of Class II are found on dendritic

cells, but can also be observed on activated
macrophages, B cells, and several other host
cell types in inflammatory conditions.
ANTIGEN PRESENTATION TO THE T CELLS

In the upper pathway; foreign protein or antigen (1) is taken up by an

antigen-presenting cell (2). The antigen is processed and displayed
on an MHC II molecule (3), which interacts with a T helper cell (4). In
the lower pathway; whole foreign proteins are bound by membrane
antibodies (5) and presented to B lymphocytes (6), which process (7)
and present antigen on MHC II (8) to a previously activated T helper
cell (10), spurring the production of antigen-specific antibodies (9).
 Clonal selection
• In the 1950s, before extensive knowledge about
protein structure, two major hypotheses were put
forth to explain the specificity of antibodies.

• The instructive hypothesis proposed that antigen

itself directed the folding of the polypeptide
chains of the antibody molecule by serving as a
template to which the antibody became adapted,
thus yeilding an antibody of high affinity.
• From a theoretical point of view, an
advantage of the instructive mechanism for
determining specificity was that only a few
different polypeptide chains would be needed
to generate the tremendous number of
antigen-binding specificities an organism
requires.
• The alternative hypothesis , termed clonal
selection, proposed that antibody receptors of
diverse specificity pre-existed on the surface of
clones of lymphocytes.
• The role of antigen was to select, bind to and
subsequently activate those clones of
lymphocytes bearing complementary
receptors.
• Clonal selection presumed that antibody structure is
determined solely by the primary structure (amino acid
sequence of) of the heavy and light chains.

• In contrast to the instructive mechanism, clonal

selection requires that each organism be capable of
producing an extremely large number of different
antibody molecules (B-cell receptors), as well as a large
number of T-cell receptor (TCR) molecules, to be able to
bind any antigen that might subsequently be
encountered in its environment.
 Structural basis for receptor
specificity
• As mentioned previously, the ability to detect
and to respond to a foreign molecule depends
on the presence of specific receptors on the
surface of lymphoctes.

• Only lymphoctyes possess receptors that

1)have affinity for antigen and
2)encompass a diverse range of specificities.
• All other cell types that participate in immune
responses, including macrophages and other
APCs, are incapable of this recognition
function.
• Thus although Class II molecules on APCs bind
antigen fragments, they lack significant
diversity and their binding is a low-affinity
interaction.
• The antigen receptor on B lymphocytes has long been
identified as an antibody molecule, which is identical in
binding specificity to the antibody eventually produced
in large quantities and secreted into body fluids by B
cells and plasma cells.

• The structure of the antibody receptor differs from

secreted antibody in that the receptor possesses a
transmembrane portion that transmits signals to the
interior of the cell after antigen binding.
• The basic unit structure of the antibody molecules
consists of four polypeptide chains, including 2
identical light chains(25,000 D) and 2 identical
heavy chains(50,000 D).
• These chains are usually linked together by
disulfide bonds, yielding an antibody of
approximately 150,000 D.
• The arrangement of the heavy and light chains
produces a Y-shaped , three dimensional
structure.
• For heavy and light chains, around 100 amino-
terminal amino acid residues contain significant
amounts of sequence variability and hence
constitute the variable (V) region of the molecule.

• The remainder of each chain to the carboxy-

terminus (100 amino acids for the light chains,
300 amino acids for the heavy chains) is constant
in sequence (constant [C] region) for each chain
type.
Variable and constant regions of
antibody
• Most of the variability in the V region of heavy
and light chains is confined to the so called
hypervariable regions.

• Hypervariable regions, also called

complementary –determining residues are
involved in the interaction with antigens.
• The side chains of the amino acid residues that
comprise the combining site determine the
nature of the interaction(s) that a particular
molecule can undergo in binding to an
antigenic determinant.
• Thus amino acid side chains may be
hydrophobic(valine, alanine) , positively or
negatively charged and so forth.
• The array of different shapes , charges and
hydrophobicities gives each combing site a
unique structure, thus permiting it to bind a
limited range of complementary structures
with significant affinity.
• For antibodies, this affinity must exceed 10 4 M
to be considered specific binding.
• T cells have their own unique antigen receptor.

• Like antibody, this molecule consists of two chains ,

termed α(42,000 D) and β(46,000 D), or in a minority of
cells, two homologous chains termed γ and δ.

• The α and β chains also possess regions that are

constant , variable, and hypervariable in sequence,
with the juxtaposition of hypervariable regions again
responsible for formation of the antigen binding site.
• However, in contrast to antibody receptors on
B cells, TCR molecules do not directly bind
antigen in solution but rather interact with the
processed fragments of antigen that are first
bound to histocompatibility molecules.

• T cell activation occurs only after formation of

this trimolecular complex (TCR, antigen,
histocompatibility molecule).
 Primary and Secondary immune
responses
• After first exposure to an antigen, antibodies
appear in the serum and other secretions only
after a delay of a week or more.
• This primary response is of short duration(weeks)
and is characterized by the production of low-
affinity antibodies of the IgM class.
• In contrast, re-exposure to the same or a closely
related antigen elicits a much more vigorous
secondary response.
• Secondary responses are long lasting and are
characterized by antibodies that are
predominantly of the IgG, IgA, or IgE class.
• These antibodies are also usually of much higher
affinity.
• This adaptive change is the consequence of the
proliferation of B-cell clones bearing receptors
able to bind, and to become activated by, a given
antigen during the primary response.
• Before antigen exposure, the number of such B
cells is extremely low; this number may
increase 1000-fold, resulting in an expanded
population of memory cells, which are then
available to mount a faster and more vigorous
response on a second exposure.
• T cell mediated primary and secondary
responses are also observed.
• The ability to mount a faster, more vigorous
secondary response depends on clonal
selection by antigen, followed by clonal
expansion.
Clonal expansion
Hematopoiesis
Cytokines affecting hematopoiesis
IL-3
• It acts as a colony stimulating factor (CSF) for
bone marrow cells.
• IL-3 stimulation of normal bone marrow leads to
proliferation of a large variety of clonal cell
populations, including colonies of granulocytes,
macrophages, megakaryocytes , eosinophils,
basophils, mast cells, and in the presence of
erythropoietin, even normoblasts and
erythrocytes.
 GM-CSF
• Granulocyte monocyte colony stimulating
factor is a glycoprotein produced by activated
T cells, monocyte/macrophage, vascular
endothelial cells, and fibroblasts.
• GM-CSF is a potent activator of macrophage
function, including induction of tumoricidal
activity and superoxide generation.
 Monocyte/macrophage CSF (M-CSF)
• This molecule exerts its effects primarily on a
stem cell that is already committed to
differenciate into monocytes.

Granulocyte CSF (G-CSF)

• It is a polypeptide produced by endothelial cells ,
fibroblasts and macrophages.
• This factor acts on marrow stem cells already
committed to development into granulocytes.
 IL-7
• Produced by the bone marrow stroma, this
molecule is intimately involved in the generation
of immature B and T lymphocytes.
• The molecule produced by the stromal cells that
promoted pre-B cell growth was termed
lymphopoietin 1, based on its capacity to
influence early lymphopoiesis.
• The factor responsible for pre-B proliferation was
renamed IL-7.
• The activity of the molecule on B cells appears
limited to promotion of the growth of
immature pre-B cells from bone marrow.
• However, IL-7 also triggers the proliferation of
immature T cells.
• It appears to be a most potential stimulator of
proliferation of the most immature of the T
cell population (i.e. , those thymocytes devoid
of expression of either CD4 or CD8).
 Mature cells from Myeloid
progenitors
• Cells that produce defined effects are called
effector cells for those activities, while cells
that support execution of the activities are
called accessory cells.
• Effector cells for inflammation and for
adaptive immunity share a common precursor,
and the inflammatory cells often play crucial
roles in immune activities.
• A cell capable of differenciation into many
different cell types is called pluripotential.
• The pluripotential cell leading to both
inflammatory and immune effector cells is
called hematopoietic stem cell.
• Progeny of the hematopoietic stem cell
differenciate into erythrocytes, leukocytes,
megakaryocytes, and phagocytic cells.
• The hematopoietic stem cell differentiates into
precursor cells that are also pluripotent.
• The lymphoid precursor cell gives rise to
several populations of lymphocytes that are
the effector cells of adaptive immunity.
• The myeloid precursor differentiates into many
cells that perform nonspecific inflammatory
activities.
• All descendants of the hematopoietic cell have
substantial capacity for cell division and
manifest continuous growth and
multiplication throughout life in the bone
marrow and other tissue sites.
• Two principal types of inflammatory cells
evolve from the myeloid precursor :
granulocytes and mononuclear phagocytic
cells (i.e. monocytes, macrophages,
histiocytes).
• Granulocytes have cytoplasmic granules that
contain enzymes. They are described as
neutrophils , eosinophils or basophils
according to their staining characteristics.
• The blood phagocytes include the neutrophil and the blood
monocyte.

• Both neutrophils and monocytes can move through tissues

or across surfaces where they engage in phagocytosis when
their membranes are suitably stimulated; and they
synthesize a wide variety of proteins , many of which are
enzymes capable of cleaving a broad range of proteins.

• However, they differ in that neutrophil is a short live cell ,

whereas the monocyte generally develops into a tissue
macrophage.
 Polymorphonuclear leukocytes and
phagoctyosis
• PMNs are a heterogenous group of cells.
• The mature form has a diameter of 10 to 15
μm and usually contains an irregularly shaped,
multilobed nucleus and cytoplasmic granules.
(On the other hand, monocytes have a large
nucleus and abundant cytoplasm that may
have a granular appearance but lack the
specific granules that distinguish
granulocytes).
• PMNs represent 60 – 70% of the total WBCs in
humans and are the hallmark of acute
inflammatory response for providing a primary
nonspecific internal defense mechanism.
• PMNs are involved in nonspecific immune
responses by :
• 1) Adherence / attachment to damaged
epithelium.
• 2) Locomotion via ameboid movement.
• 3) Diapedesis through the wall of a blood vessel.
• 4) Chemotaxis toward the particles to be
engulfed.
• 5) Phagocytosis of the particles.
• 6) Increased metabolism through oxidative burst
and glycolysis.
• 7) Degranulation .
• 8) Digestion of the foreign material.
 Adherence
• Adherence of PMNs to the vascular
endothelium is not complete in the beginning;
however, these cells become sticky after
exposure to chemotactic factors.
• One major contact system requires
combination between a molecule on the
leukocyte membrane and a complementary
ligand on target cells.
• Granulocytes, lymphocytes and some
macrophages express a leukocyte function-
associated antigen1 (LFA-1) which binds the
intercellular adhesion molecule 1 (ICAM-1)
that exists on endothelial cells, certain APCs
and many mucosal cells.
• Blocking of these molecules can inhibit T-cell
cytotoxicity, NK –cell cytotoxicity and antibody
dependent cell-mediated cytotoxicity (ADCC).
• LFA-1 is also expressed on developing B cells
at the pre-B or later stages and on
macrophages stimulated by IFN-γ and other
mediators , but it does not occur on
nonhematopoietic cells.
• Expression of ICAM-1 is increased by acute
inflammatory events and by the products of
activated lymphocytes and macrophages,
specifically IL-1 and IFN-γ.
 Locomotion
• Once bound to the endothelial cells, locomotion
of PMNs is similar to the movements of amebae.
• These cells can move on the surfaces of blood
vessel walls, changing direction by expressing
pseudopods from a different part of the cell
surface.
• In the absence of specific stimuli, this locomotion
is not in a straight line and is undirected.
 Diapedesis
• Diapedesis or emigration through the wall of
the blood vessel occurs following adherence to
the endothelium.
• The PMN inserts psedopods between the
endothelial cell junctions , disrupting the
basement membrane at the location so that
the cell can squeeze itself into the juxtaposed
tissue spaces.
 Chemotaxis
• Chemotaxis is the process by which phagocytes
are attracted to sites of inflammation.
• Directed migration of the PMNs is mediated
primarily by soluble components of the
complement system, particularly C5a.
• Both neutrophils and macrophages are attracted
by C5a, but neutrophils are the predominant cell
in sites of acute inflammation.
• Other chemotactic factors include certain
products of coagulation or fibrinolytic pathways
and certain bacterial products.
 Phagocytosis
• Phagocytosis is a process whereby the
external cell wall of the PMN adheres to and
completely surrounds the offending bacterium
or other particle, encapsulating the foreign
substance with a layer of reversed membrane
called a phagosome.
• Once they have arrived at the site of
inflammation, the phagocytes must recognize
the infectious agent .
• Receptors on their surface allow them to
attach nonspecifically to a variety of
microorganisms.
• Attachment is greatly enhanced if the
microorganism has been opsonized by IgG
and/or the C3b component of complement.
• C3b and IgG function as opsonins because PMNs
and other phagocytic cells have receptors on their
surface that specifically recognize the exposed
portions of those molecules when they coat
bacteria or other particles.
• After attachment the phagocytes engulf the
microorganism by extending psedopodia around
it. These fuse and the microorganism is
internalized in the phagosome.
• Subsequently, fusion of the phagosome with
enzyme containing granules (phagolysosome)
and degradation of the bacteria by the enzymes
in the phagolysosome occurs.
• The mature PMN has distinct granules that
contain hydolases, myeloperoxidase , lysozyme ,
lactoferrin and cationic proteins.
• Each of these biochemical components either
performs a bactericidal function or degrades the
organic materials that remain after the bacteria
are killed.
 Increased metabolism
• During phagocytosis the metabolism of
leukocytes changes rather dramatically as O2
and glusose are consumed , primarily through
the hexose monophosphate shunt.
• H2O2 is also produced during the respiratory
burst that occurs as the phagocytosed particle
is digested.
 Degranulation
• Degranulation of PMNs involves the fusion of
intracellular granules with the plasma membrane.
• The contents of the granules are then released
either into the phagosome or into the
accumulated extracellular fluid.
• As in inflammation , much of the tissue damage
associated with injury is caused by the contents
of the granules that are released during the
degranulation of PMNs.
 Digestion
• Digestion occurs following degranulation.
• When the granules are fused with the vacuole
containing the ingested particle, the vacuole is
exposed to the lytic action of the enzymes.
• The mechanism by which many types of
bacteria are killed following ingestion by
phagocytes requires H2O2 and the enzyme
myeloperoxidase.
Neutrophils
 Neutrophils
• Neutrophils represent about 70% of circulating
nucleated cells.

• Their main function is phagocytosis; since Fc

and complement receptors are expressed on
cell surface, their phagocytic activities are
enhanced when particles or antigens are
coated with antibody and/or
complement(opsonization).
• The first cells to arrive at an inflammatory
focus are neutrophils, which appear within a
few hours.
• Initially, neutrophil development is a
proliferative phase; with cell division, the cells
evolve from the myeloblast to promyelocyte
and then to the myelocyte.
• During this period, neutrophils acquire their
characteristic granules.
• The final phase of development in the bone
marrow is maturation with no cell division.
• The bone marrow of a healthy adult produces
more than 1011 neutrophils per day.
• On release from the bone marrow into the
circulation, the cells are in a nonactivated state
with a half life of 6 hrs.
• Within the circulation, the cells become activated
by chemoattractants, with dramatic alterations in
their surface characteristics.
• Receptors for chemoattractants increase
twofold to fourfold , and receptors for
immunoglobulins and complement
components (C3b) become mobilized from
intracellular compartments , resulting in a
primed cell that has increased adherence and
avidly migrates to endothelial sites.
• Localized margination takes place in areas of
high concentration of chemoattractants.
• Chemotaxis and diapedesis ensues.
• Diapedesis is accompanied by an orderly release
of neutrophil granules and generation of toxic
oxygen products.
• The cytoplasmic primary or azurophilic granules
contain enzymes including hydrolase,
myeloperoxidase and muraminidase.
• The primary granules function predominantly in
the intracellular milieu in the phagolysosomal
vacuoles where they are involved in the killing
and degradation of microorganisms.
• Myeloperoxidase is a critical enzyme in the
conversion of hydrogen peroxide to
hypochlorous acid.
• Primary granules lysosomal proteases such as
elastase and collagenase, cathepsin G and
gelatinase can degrade proteins following the
alteration of their tertiary structures by
reactive oxygen products.
• Collagen breakdown products have
chemotactic activity for neutrophils,
monocytes and fibroblasts.
• Regulation of tissue destructive potential of
lysosomal products is mediated by protease
inhibitors such as α2 macroglobulin and α1
antitrypsin .
• These antiproteases are present in serum and
synovial fluids.
• Neutrophil secondary granules contain lactoferrin
and vitamin B12 binding protein.
• Unlike the primary granules, neutrophil
secondary granules are particularly susceptible to
extracellular release.
• These granules are considered the intracellular
reservoir of various plasma membrane
components including receptors for C3b and
laminin, plasminogen activator and alkaline
phosphatase.
• Neutrophil specific granule hydrolases also
produce mediators of inflammation through
mobilization of monocytes by a monocyte
chemoattractant and through their direct
action on complement components such as C5
and kininogen to liberate C5a and kinin ,
respectively.
• On activation, neutrophils and mononuclear
phagocytes have increased oxygen
consumption , a process known as respiratory
burst.
• Oxygen is univalently reduced to superoxide
anion or its protonated form(HO2-), which then
is catalytically converted by action of
superoxide dismutase to H2O2.
• The superoxide anion (O2-) is both a one-
electron reductant and a one –electron
oxidant that can pass through cell membranes
via anion channels.
• It appears that the superoxide anion does not
have direct toxic effects on targets but rather
exerts its toxicity by penetration to important
sites where it subsequently is converted into
other toxic products.
• H2O2 can be produced directly from the
divalent reduction of oxygen by glucose
oxidase or by the intermediate formation of
O2- by xanthine oxidase.
• H2O2 interacts with myeloperoxidase , which is
stored in the primary granules , to produce
hypochlorous acid, which is metabolized to
hypochlorite and chlorine.
• The hyroxyl radical (OH-) and hypochlorite are
products of oxygen metabolism generally
thought to be important in cytotoxic reactions.
• The toxicity of OH- is believed to result from its
ability to serve as a powerful oxidant
capturing electrons from a large variety of
compounds with the formation of a new
radical, which can then oxidize other
substances.
• New data from the Max NETs
Planck Institute for Infection
Biology in Berlin show that
neutrophils also have
another defense
mechanism.

• They can "spit out" a net-

like structure that binds
bacteria, disarms and finally
kills them. These structures
are called NETs (Neutrophil
Extracellular Traps).
 Eosinophils
Eosinophils
 Eosinophils
• Eosinophils (or, less commonly, acidophils), are
white blood cells that are one of the immune
system components responsible for combating
infection and parasites.
• Along with mast cells, they also control
mechanisms associated with allergy and asthma.
• They are granulocytes that develop during
haematopoiesis in the bone marrow before
migrating into blood.
• In normal individuals eosinophils make up about 1-6%
of white blood cells, and are about 12-17 micrometers
in size.

• They are found in the medulla and the junction

between the cortex and medulla of the thymus, and, in
the lower gastrointestinal tract, ovary, uterus, spleen,
and lymph nodes, but not in the lung, skin, esophagus,
or some other internal organs under normal conditions.

• The presence of eosinophils in these latter organs is

associated with disease. Eosinophils persist in the
circulation for 8-12 hours, and can survive in tissue for
an additional 8-12 days in the absence of stimulation.
• Eosinophils develop and mature in bone
marrow. They differentiate from myeloid
precursor cells in response to the cytokines
interleukin 3 (IL-3), interleukin 5 (IL-5), and
granulocyte macrophage colony-stimulating
factor (GM-CSF).
• Eosinophils produce and store many secondary
granule proteins prior to their exit from the
bone marrow.
• After maturation, eosinophils circulate in
blood and migrate to inflammatory sites in
tissues, or to sites of helminth infection in
response to chemokines like CCL11 (eotaxin-
1), CCL24 (eotaxin-2), CCL5 (earlier called
RANTES for Regulated upon Activation,
Normal T cell expressed and secreted), and
certain leukotrienes like LTB4.

• At these infectious sites, eosinophils are

activated by Type 2 cytokines released from
a specific subset of helper T cells (Th2).
• Following activation, eosinophil effector functions
include production of:
• cationic granule proteins and their release by
degranulation.
• reactive oxygen species such as superoxide.
• lipid mediators like the eicosanoids from the
leukotriene and prostaglandin families.
• enzymes, such as elastase
• growth factors such as TGF beta, VEGF, and PDGF
• cytokines such as IL-1, 2, 4 ,5,6, 8, 13 and TNF alpha.
• In addition, eosinophils play a role in fighting viral
infections, which is evident from the abundance
of RNAses they contain within their granules, and
in fibrin removal during inflammation.
• Eosinophils along with basophils and mast cells,
are important mediators of allergic responses
and asthma pathogenesis and are associated
with disease severity.
• They also fight helminth colonization and may be
slightly elevated in the presence of certain
parasites.
• Eosinophils are also involved in many other
biological processes, including postpubertal
mammary gland development, allograft
rejection and neoplasia.
• They have also recently been implicated in
antigen presentation to T cells.
• Following activation by an immune stimulus,
eosinophils degranulate to release an array of
cytotoxic granule cationic proteins that are
capable of inducing tissue damage and
dysfunction. These include:
• Major basic protein (MBP)
• eosinophil cationic protein (ECP)
• eosinophil peroxidase (EPO)
• eosinophil-derived neurotoxin (EDN)
• Major basic protein, eosinophil peroxidase,
and eosinophil cationic protein are toxic to
many tissues.
• Eosinophil cationic protein and eosinophil-
derived neurotoxin are ribonucleases with
antiviral activity.
• Major basic protein induces mast cell and
basophil degranulation, and is implicated in
peripheral nerve remodelling.
• Eosinophil cationic protein creates toxic pores in the
membranes of target cells allowing potential entry of
other cytotoxic molecules to the cell, can inhibit
proliferation of T cells, suppress antibody production by
B cells, induce degranulation by mast cells, and
stimulate fibroblast cells to secrete mucus and
glycosaminoglycans.

• Eosinophil peroxidase forms reactive oxygen species

and reactive nitrogen intermediates that promote
oxidative stress in the target, causing cell death by
apoptosis and necrosis.
 b

Basophils
 Basophils and Mast cells
• Cells that participate in immunologic reactions
by release of biochemical substances
(mediators) include mast cells, basophils and
platelets.
• The biological activities of these mediators
include increased vascular permeability,
smooth muscle contraction and augmentation
of the inflammatory response.
• Human skin and g.i. tract are particularly rich
in mast cells .
• Granules containing the potent mediators(eg.
Histamine, heparin, serotonin , hyaluronic acid
and eosinophil chemotactic factor of
anaphylaxis[ECF-A] ) are released from mast
cells on injury to these tissues.
• Immunological reactions involving IgG or IgE,
which bind to receptors on the surface of mast
cells and basophils can trigger degranulation
and release of the mediators into the
circulation.
• Basophilic granulocytes
make upto 0.5% to 2% of
circulating leukocytes.

• At one time it was

thought that basophils
were circulating mast
cells, but studies have
shown that these two cell
types differ in structure
and content of their
granules.
• Basophilic granules contain primarily histamine and
leukotrienes LTC4, LTD4 and LTE4 (SRS-A) , which are
potent spasmogenic agents causing smooth muscle
constriction.
• Both basophils and mast cells are characterized by
deep violet blue cytoplasmic granules that contain
vasoactive amines like histamine, both express Fc
receptors for IgE heavy chain such that IgE antibodies
bind to these cells and remain stable; both respond
when IgE and antigen immune complexes bind to the
IgE receptors on the basophils to stimulate
degranulation.
• The primary function of basophil appears to be
amplification of the reaction that starts with
mast cells at the site of initial tissue disruption.

• In spite of similarities, basophils and mast cells

are distinct cell populations in that:
 Basophils Mast cells
• Derived from • Mesenchymal origin.
myeloblast like cells.
• Circulate in peripheral • Widely distributed in
blood. the tissues (thymus,
spleen, lung, g.i.t , bone
marrow).
• Granules which are finer
than those of basophils.
Platelets
 Platelets
• Platelets contain heparin, serotonin and
lysosomal enzymes that are released from the
granules during platelet aggregation and that
may contribute to the acute vascular phase of
inflammation.
• Platelets may also produce oxygen radicals,
which may cause tissue damage.
• However the main role of platelets is to block
damaged vessel walls and prevent hemorrhage.
Platelets initiating a clot and covered
in fibrin meshwork
• Platelets are derived from megakaryocytes in
the bone marrow and contain granules.

• They are also involved in the immune

response, especially inflammation.

• They possess class I MHC product, receptors

for IgG and low affinity receptors for IgE.
• In addition, platelets carry receptors for
fibrinogen and von Willebrand factor binding.

• Following endothelial injury, platelets adhere

to and aggregate at the endothelial surface
releasing permeability-increasing substances
and factors responsible for activating
complement components to attract
leukocytes.
Monocytes &
Macrophages
 Monocytes and macrophages
• An organism that successfully penetrates the
epithelial surface encounters phagocytic cells
of the monocyte/macrophage lineage.

• These cells are derived from the bone marrow

stem cells.
• Myeloid progenitors from the bone marrow
give rise to cells of the mononuclear
phagocyte system.
• Monocytes represent about 4-10% of the
circulating nucleated cells.
• They have a diameter of 12 to 16 μm. The
nucleus is horseshoe shaped and the
cytoplasm contains neutrophilic granules.
• Circulating monocytes can migrate into
various tissues and become tissue
macrophages.
• Circulating monocytes represent cells that
have not fully differentiated; further evolution
occurs at various tissue sites at which the
monocytes are deposited.
• Differentiated cells of this series include
macrophages (lymph nodes, peritoneal lining,
pleural surfaces , joint linings, renal glomeruli
and inflammatory tissues), dendritic cells
(lymph nodes), Kupffer cells(liver), Langerhans
cells(skin), alveolar macrophages( lungs) ,
osteoclasts(bone), microglia (CNS), and
histiocytes (connective tissue).
• These cells play a pivotal role in both cellular
and humoral immunity.

• These cells have several cytoplasmic

enzymes(nonspecific esterase, lysozyme) ,
they have a consistent occurance of
membrane receptor molecules like receptors
for IgG molecules(the Fc receptor) and the
complement C3 receptor, they have a
capacity to phagocytize particles coated with
Ab or complement proteins(particles that
have been opsonized) , and they also have the
ability to directly phagocytize and kill many
microorganisms, particularly after activation
by lymphokines.
• They also process and present antigen for
subsequent B lymphocyte development and
antibody synthesis.
• The term reticuloendothelial system (RES)
designates all actively phagocytic cells.
• Debris removed by the mononuclear
phagocytic system (MPS) include debilitated
erythrocytes, leukocytes and platelets ,
bacteria and antigen-antibody complexes.
• When the MPS is actively eliminating debris
form the circulation, organs that are
populated by tissue macrophges exhibit
lymphadenopathy (enlarged lymph nodes),
splenomegaly or hepatomegaly.
• Compared with circulating monocytes,
macrophages have a wider repertoire of
hydrolytic enzymes and greater phagocytic
capacity.
• Tissue macrophages retain some capacity to multiply
after extensive stimulation ; however, macrophage
populations are generally replenished from the bone
marrow.
• Following the inflammatory stimulus of the RES, the
macrophages that accumulate constitute the exudate
macrophages.
• These cells synthesize and secrete cytokines that
activate cells of the immune system and influence
other aspects of inflammation, complement activity
and multiplication of cells that generate new tissue and
and repair damaged tissue.
• Mononuclear phagocytes respond to the same
chemoattractants that attract PMNs (eg. C5a);
however, the primary chemoattractants to
which the macrophages respond are soluble
factors from T lymphocytes such as IFN-γ.
• Peripheral blood monocytes are much less
efficient at phagocytosis and killing bacteria
than are neutrophils.
• Circulating PMNs are end stage cells; i.e. after
phagocytosis and degranulation, they die.
• Macrophages, on the other hand, appear to
be stimulated by the processes involved with
phagocytosis . They become secretory cells ,
synthesize acute phase proteins, and may
even proliferate locally within tissues.
• When macrophages are stimulated by
phagocytic events, they become secretory
cells, that produce and secrete IL-1.
• IL-1 activates T cells and a range of other
factors that non specifically regulate the
activities of lymphocytes, including IFN, PGs,
complement components and acute phase
proteins.
• Finally, macrophages are responsible for
antigen recognition.
• Antigen presentation is an important function
of a subset of macrophages that have HLA-DR
antigens on their surface.
• The HLA-DR(MHC class II) molecule appears to
interact with the antigen and allows the
macrophage to communicate with and
synthesize antigen- specific T lymphocytes.
• Thus macrophages overlap as effectors of both
natural and adaptive immunity; they can be
involved in inflammatory responses and
antigen presentation ; and once activated by
lymphokines, they demonstrate nonspecific
cytotoxicity to altered cells by both direct
contact and cytolytic factors(i.e. TNF-α).
 Mature cells from lymphoid
precursors
• Lymphocytes descend from the lymphoid
precursor; they characteristically have a
round, dense nucleus and less cytoplasm.

• Two major subpopulations, the T lymphocytes

and B lymphocytes exist.
• Lymphocytes are produced in the primary
lymphoid organs such as bone marrow and
thymus.
• Approximately 20-30% of circulating nucleated
cells are lymphocytes.
• These lymphocytes can be classified into two
categories.
 Large granular
Small lymphocytes lymphocytes(LGLs)
• Have a diameter of 8 to 10 • Have a diameter up to 16
µm . µm.
• Have a higher nuclear-to- • Have a smaller N/C ratio.
cytoplasm (N/C) ratio.
• Lack cytoplasmic granules. • Contain cytoplasmic
• Further divided into two granules.
major classes: T and B
lymphocytes on the basis of
their functions.
• Individual lymphocytes are specialized to
respond to a limited group of structurally
related antigens.

• This specificity of response is related to

membrane receptors on the cells that
recognize the antigen.
• Lymphocytes express a large number of
phenotypic markers on their surface.
• Molecules that can be used to distinguish cell
populations can be identified by specific
monoclonal antibodies.
• The term cluster determinant(CD) is used to
indicate the specific molecule recognized by a
group of monoclonal antibodies.
 Morphogenic heterogeneity
• Resting blood T cells show two characteristic
morphological patterns.

• The majority of TH(helper) and TC/S (supressor

or cytotoxic) cells carry a cytoplasmic Gall
body, which consists of a group of primary
lysosomes.
• Resting blood B cells do not display Gall bodies
or granular lymphocyte morphology, and their
cytoplasm is predominantly occupied by single
ribosomes.

• Occasionally activated B cells are found with a

rough endoplasmic reticulum.
• Third population cells(TPCs) are characterized
by granular lymphocyte morphology.
• In contrast to granular T cells, they display a
large number of azurophilic granules.
• An additional type of lymphocyte mediates
more nonspecific type of activities and
includes NK cells as well as K cells involved in
ADCC(Antibody dependant cell mediated
cytotoxicity) reactions.
• Human B lymphocytes are exposed very early
to maturational effects of liver elements.
• B cells undergo gene rearrangements that
confer the ability to manufacture Igs.
• Activation and subsequent differentiation
cause antigen stimulated B lymphocytes to
evolve into plasma cells, which secrete the
antibody into body fluids.
• After activation, B lymphocytes evolve either
to short-lived antibody producing plasma cells
or long lived memory cells that can rapidly
multiply, but are not actively synthesizing
antibody.
• T cells develop in the thymus, whereas B cells
differentiate in fetal liver and in the adult
marrow in mammals.
• TPCs (called non-T, non-B cells or “null cells”)
also develop in bone marrow.
• This TPC population contains cytotoxic
lymphocytes (NK cells), a variety of cells
specialized for presenting antigen to T and B
cells (i.e. , APCs) and platelets.
• Lymphocytes are produced in the thymus and
adult bone marrow.
• Some of these cells migrate via the circulation
into the spleen, the lymph nodes, the tonsils,
and unencapsulated lymphoid tissue.
• Lymphoid cells represent about about 20% of
the total WBC count.
T lymphocytes
 T cells
• Developing lymphocytes assume the
properties of T cells as they pass through the
thymus; this generates enough committed
precursor cells such that T-cell levels can be
maintained for life, long after the organ
disappears.
• Uncommitted cells enter the thymus at the
outermost layer of cortex; cells of increasing
maturity are seen toward the interior.
• Cells within the thymus are called thymocytes
and are not fully mature.

• Fully instructed cells that have left the thymus

are described as peripheral T cells.

• Of the circulating lymphocytes, 60% to 80%

are T cells.
• The T lymphocytes are associated with two
types of immunological functions, effector and
regulatory.
• The effector functions include activities such as
killing of virally infected cells and tumors.
• The regulatory functions are represented by
their ability to amplify or suppress through
cytokines or other effector lymphocytes,
including B and T cells.
• Subsets of TH and Tc/s cells are involved in
regulating immune responses, cooperating
with B cells in the induction of antibody
synthesis, releasing lymphokines that can
activate macrophages, and aiding in
elimination of many intracellular and viral
pathogens.
• Most circulating T cells express three of the
following CD markers:
• 1) CD2 receptor for sheep erythrocytes
• 2) CD3 T-cell antigen receptor complex
• 3) CD4 receptor for MHC class II molecule
• 4) CD8 receptor for MHC class I molecule
• The bone marrow T stem cells contain the
enzyme TdT but do not express surface CD
glycoproteins.
• The earliest differentiation event is acquisition of
CD2 and CD3 , which occurs in the thymic cortex.
• Further differentiation in the cortex occurs as the
T cells move through the thymic medulla:
• 1)development of two distinct T-cell populations,
helper-inducer T cells (CD4 positive) and
suppressor-cytotoxic T cells (CD8 positive), and
• 2)loss of TdT enzyme.
• The most significant outcome of this
intrathymic differentiation is that the T cells
learn to recognize self-MHC gene products,
which are essential for cellular interaction in
the immune responses.
• The mature T lymphocytes then seed
peripheral lymphoid organs from the
bloodstream.
• Although one of the first ways to distinguish
human T cells from B cells is by their ability to
bind to sheep erythrocytes via the CD2 molecule,
the definitive T cell marker is the T cell antigen
receptor (TCR).
• Presently two types of TCR are defined: TCR-2 is
a heterodimer of two polypeptides, α and β; TCR-
1 is structurally similar but consists of γ and δ
polypeptides.
• Both receptors are associated with a complex of
polypeptides making up the CD3 complex.
• Approximately 95% of blood T cells express
TCR-2, and upto 5% have TCR-1.
• The TCR-2 bearing cells can be subdivided further
into two distinct populations: the TH subset which
is CD4+ and the Tc/s subset which is CD8+.
• CD4+ T cells recognize antigens in association
with MHC class II molecules , whereas CD8+ T
cells recognize antigens in association with MHC
class I molecules.
• The CD4+ set can be further divided
functionally into: 1) cells that positively affect
the immune responses of T and B cells,
representing the helper cell function; and
2) cells that activate suppressor/cytotoxic
functions of CD8+ cell, representing a
suppressor/inducer function.
• T lymphocytes recognize their specific antigens
through a complex of membrane proteins
called the T-cell receptor, which has 2 parts:
TCR and CD3.
• TCR identifies the idiotype of the specific
antigen.
• CD3 is present on all T cells and probably
conveys to the cell interior the message that
TCR has combined with antigen.
• T cells recognize antigen only when the APC
offers class I or II MHC products in association
with the antigen.
• Recognition of the MHC products depends on
a receptor molecule closely associated with
the TCR heterodimer.
• B lymphocytes and the other antigen
presenting cells that process and present
antigen consistently express both class I and
II material and can interact with both T cell
populations.
• Somatic cells express only class I products;
thus they are potential targets for cytotoxic
attack.
• After activation, T cells express receptors
different from those of resting T cells.
• When the CD3 complex interacts with antigen,
the receptor-ligand complex is internalized,
leaving few antigen receptors on the cell
surface.
• In its place a new molecule appears, called Tac
or CD25.
• This receptor is an activation marker and is a
receptor for IL-2.
• Once binding of antigen by CD3 occurs, there is
stimulation by IL-1, and the antigen-stimulated T
cell expresses CD25 and begins producing IL-2.
• MHC class II molecules that characterize
lymphoid precursor cells, B lymphocytes and APCs
become detectable on activated T cells after their
exposure to antigen and IL-1.
• Although T cells as a population appear to be
capable of synthesizing many lymphokines,
individual T cells produce a limited repertoire of
individual lymphokines.
• CD4+ cells release IL-2 and IFNγ.
• Another subpopulation of CD4+ cells secretes only
IL-4 or IL-5.
• These subsets have been described as Th1 or Th2
cells, with Th1 largely involved in delayed type
hypersensitivity reaction and Th2 cells responsible
for T-B cell cooperation.
B lymphocytes
 B cells
• B lymphocytes make up approximately 5% to
15% of circulating nucleated cells of normal
human blood.
• They are classically identified by their cell
surface Ig (sIg).
• The sIg is endogenously synthesized by the B
cells and serves as the antigen receptor.
• Developmentally all B cells initially express
membrane IgM.
• Addition of IgD occurs on most B cells ; this
isotype is unique in that it is rarely secreted.
• The majority of resting B cells are IgM+ and
IgD+ , whereas IgD tends to be lost soon after
activation.
• In mucosal-associated lymphoid tissues(MALT),
the majority of B cells express IgA as their sIg.
• Other B cell markers are Fc receptors for the IgG
heavy chain, for complement components (C3b
and C3d), for MHC class II molecules, and for the
Epstein-Barr virus that identifies the lymphocyte
surface CDs.
• The CD markers that are frequently used to
identify B cells are CD19, CD20 and CD21.
• B lymphocytes undergo instruction and
differentiation in the bone marrow and
possibly also in the fetal liver.
• Small resting B cells bear surface Ig molecules.
• Each of these is a heterodimer of 2 heavy and
2 light chains, and each chain is composed of
variable and constant region domains.
• Rearrangement of Ig genes signals irreversible
commitment to the B cell lineage.
• Synthesis of heavy chains begins in those cells
where rearrangement was successful, yeilding
lymphocytes that can be distinguished as large
pre-B cells.
• The pre-B cells are thought to reduce in size,
rearrange light chain genes and display
surface Ig.
• A low, steady state rate of shedding and
replacement of membrane Ig in B lymphocytes
occurs , but this increases dramatically on
activation.
• Plasma cells can produce and release
thousands of molecules per second.
• B lymphocytes differ in terms of specificity and
their antibody-combining sites.
• The morphology and Ig biosynthetic rates of
resting B cells and plasma cells are quite
distinct.
• Antigen experienced memory B cells differ
from virgin B cells.
• Another transient marker for early B cells is
the membrane glycoprotein formerly called
CALLA, now designated CD10.
• Lymphoid precursors express MHC class II
products, and these remain apparent on B
cells through every stage upto the plasma cell.
• The membrane of the fully developed plasma
cell lacks class II chains.
• Several receptors appear as B cells reach
functional maturity.
• These react with a structural element of the Ig
molecule Fc segment.
• Fully differentiated plasma cells engaged in
antibody production do not have Fc receptors.
• Unlike T cells, B cells have membrane
receptors for several proteins of the
complement system and the receptor for C3d ,
designated CD21, is the target molecule for
the EB virus which infects B lymphocytes.
• A very important group of molecules on B
lymphocytes are termed MHC class II antigens.

• They bind antigen fragments, and the

resulting complex is recognized on the B cell
surface by the Th cells.
• Plasma cells are specialized to manufacture and
secrete Igs.
• Membrane Ig is no longer expressed; the class II
MHC products disappear, as do the receptors for
Ig heavy chains, complement proteins, and CD
glycoproteins.
• After a few days of manufacturing and releasing
Ig molecules, the individual plasma cell (a
terminally differentiated B cell) dies without
reproducing.
• B cells that have participated in an immune
response (memory B cells) differ in average life
span from newly formed or virgin B cells.
• Distinctive characteristics of memory cells are
their long average life span.
• In contrast to memory B cells, plasma cells
have relatively limited life span and
proliferative potential.
 Natural killer cells(NK cells)
• Approximately 5% of blood lymphocytes have
neither the membrane Ig characteristics of B
cells nor phenotypes characteristic of T cells.
• Originally called TPCs or null cells , certain of
these cells are now identified as NK cells.
• NK activity is the capacity to lyse target cells,
regardless of antigenic specificity, MHC
expression , or previous exposure.
• NK cells are leukocytes that can recoginze cell
surface changes that occur in some virally
infected cells and some tumor cells.
• They bind to these target cells and kill them.
• This kind of reaction in which a lymphocyte
kills a target cell is called cytotoxicity.
• The cells responsible for NK activity are
primarily LGLs(Large granular lymphocytes).
• NK cells have on their membranes a high-
affinity receptor for the Fc portion of IgG.

• Ig molecules are more strongly bound by Fc

receptors on NK cells than on other
lymphocytes.
• NK cells attack target cells with which they have
had no previous contact.
• This cell-to-cell contact does not lead to enhanced
activation or later memory.
• NK activity does, however, increase under
conditions of heightened immune activity;
immune reactions generate cytokines that
enhance NK activity, especially IFNγ (a product of
antigenically stimulated T cells) and IL-2
(produced by T cells exposed to IL-1).
• Intimate contact with the cell membrane is
essential for NK-mediated cytolysis.
• The effect may result from proteolytic
enzymes stored in the cytoplasmic granules.
• In addition, NK cells can kill a target cell,
disengage and then repeat the process with
the next target.
• Although NK cells act against a wide range of
target cells, they display more discrimination
than do macrophages and neutrophils.
• The surface proteins with which NK cells react
include HLA antigens of all allotypes, viral
products, and membrane markers present on
immature cells of various lines.
• The less mature the cell, the more susceptible
it is to NK mediated cytolysis.
• Unlike CTLs (cytotoxic T cells), the cytotoxic
activity of NK cells is not MHC restricted.

• They play an important role in ADCC (antibody

dependent cellular cytotoxicity.
• The NK cell require no time for specific
activation; thus they provide an immediate
nonspecific response to virally altered cells.
• They also provide a bridge between innate and
acquired immunity because they can be
activated by products of immune T cells,
including IFNγ and IL-2.
 Oral immune responses
• Neutrophils are believed to play an important
role in controlling the periodontal microbiota.
• The first leukocytes to arrive at the site of
inflammation, they are the dominant cell type
within the junctional epithelium and the
gingival crevice.
• Disorders of neutrophils are associated with
invasive periodontal infection and aggressive
periodontitis.
• About 1% to 2% of neutrophils migrate across
the junctional epithelium daily.

• In vitro studies have shown that P. gingivalis

impedes the transepithelial migration of
neutrophils.
• Patients with periodontitis exhibit very high titres
of IgG to specific periodontal pathogens.
• Though B cells produce antibody, T cells are
required to regulate the isotype switch from IgM
to IgG.
• APCs are abundant within the gingival tissues and
can transport antigen to regional lymph nodes,
promoting the production of IgG.
• Local immunoglobulin production has also been
documented within the gingiva.
 Immunity in periodontal disease
Gingivitis
• Gingivitis is primarily a response to the
bacteria in plaque.
• It includes a vascular response with increased
fluid accumulation and inflammatory-cell
infiltration.
• The early response is mostly lymphocytic ,
represented by T cells; however, tissue
affected by advanced and more chronic
gingivitis can contain plasma cells.
• The predominant cell type in inflammed
gingival tissue in adolescents has been
identified as the T cell.
• These local responses are not associated with
bacteria in tissues, but are in response to
products that appear to traverse the gingival
epithelium, which has lost some of its innate
protective barrier functions.
• In experimental gingivitis , the inflammatory
lesion remains dominated by lymphocytes.
• Moreover, T cell and PMN levels peak early in
experimental gingivitis and then decrease,
which would be consistent with the B-cell
infiltrate noted in more advanced periodontitis
tissues.
 Chronic periodontitis
• Extensive studies have been performed
examining the characteristics of inflammatory
infiltrate in CP.
• The distribution of gingival mononuclear cells
has shown plasma cells(5% to 15%),
lymphocytes(30% to 35%) and macrophages
(40% to 55%) in CP.
• The plasma cells are predominated by IgG
followed by IgA. No IgM cells are found in
tissues from CP.
• Moreover, IgG cells in the gingival tissues were
identified as IgG1> IgG2 > IgG3 > IgG4 and
IgA1, with high IgA2-cell levels in advanced
lesions.
• A large number of B cell lineage cells in gingival
tissues present a phenotype that can strongly
stimulate autoreactive T cells.
• Thus, immunoregulation of local polyclonal B-cell
activation may be important in the progression of
periodontitis.
• The Th /Ts ratios are reduced in chronic
periodontitis lesions, compared to homologous
peripheral blood and tissues with chronic
gingivitis.
 In summary,
• In periodontal disease, the inflammatory
response causes tissue resistance to bacterial
invasion, but also provides mechanisms that
contribute to tissue damage.
• The acute inflammatory cells such as mast cells,
macrophages, Langerhans cells, PMNs, all
combine to form a potent antibacterial defense
mechanism, but initial signs of gingival
inflammation (eg. Swelling, redness, bleeding on
probing) represent the tissue destructive aspects
of these cells.
• In established periodontal disease, there is also
chronic inflammatory change, with B cells and T
cells adding to the antibacterial spectrum.
• These lymphocytic cells have also the capacity to
release cytokines, which may induce the synthesis
of arachidonate metabolites (especially PGE2),
and to stimulate macrophages and osteoclasts to
release hydrolases and collagenases , which are
responsible for loss of collagen and bone.
• Each patient with dental plaque has a complex
balance between these protective and
damaging scenarios that result either in a
slowly progressive loss of attachment
(periodontitis) or in a restriction of tissue
inflammation to the peripheral
tissues(gingivitis).
 References
• Contemporary oral microbiology and
immunology by Slots and Taubman.
• Carranza’s Clinical Periodontology, 10th
Edition.
• Wilson and Korman.
• Wikipedia.
 ha
Thank you!