Cells of the Immune System

Immune System
3 Major Functions

Protection from disease causing invaders

Removal of dead/damaged tissues and cells

Recognition and removal of abnormal cells

Immunologic mistakes

Incorrect responses – Autoimmunity

Overactive responses – Allergy

Lack of responses - Immune deficiency

The immune system begins to develop in the embryo and by the time the baby is born, it is a
sophisticated collection of tissues that includes the blood, lymphatic system, thymus, spleen, skin and
mucosa.
The immune system responds to any antigen, whether is it harmless, like grass pollen or harmful such
as a virus or bacterial infection. Everyone’s immune system is different and reacts differently to every
antigen.
Cell Potency

Embryonic
stem cells
Stem cells are biological cells found in all multicellular organisms, that can divide (through mitosis)
and differentiate into diverse specialized cell types and can self-renew to produce more stem cells.
In mammals, there are two broad types of stem cells: embryonic stem cells, which are isolated
from the inner cell mass of blastocysts, and adult stem cells, which are found in various tissues.
In adult organisms, stem cells and progenitor cells act as a repair system for the body, replenishing
adult tissues. In a developing embryo, stem cells can differentiate into all the specialized cells
(these are called pluripotent cells).

Cell Potency:

Totipotent: Stem cells can differentiate into embryonic and extra embryonic cell types. Such cells
can construct a complete, viable organism. These cells are produced from the fusion of an egg and
sperm cell. Cells produced by the first few divisions of the fertilized egg are also totipotent.

Pluripotent stem cells are the descendants of totipotent cells and can differentiate into nearly all
cells.

Multipotent stem cells can differentiate into a number of cells, but only those of a closely related
family of cells.

Oligopotent stem cells can differentiate into only a few cells, such as lymphoid or myeloid stem
cells.

Unipotent cells can produce only one cell type, their own.
Differentiation of Human Tissues.
Our bodies have many adult stem cell niches

Blood

Gut

Skin

Bone, Skin, Tendon,

cartilage

Liver
Megakaryocytes- Platelets
SCF= Stem Cell Factor Tpo= Thrombopoietin
IL= Interleukin GM-CSF= Granulocyte Macrophage-
colony stimulating factor
Epo= Erythropoietin M-CSF= Macrophage-colony
stimulating factor
G-CSF= Granulocyte-colony stimulating factor
SDF-1= Stromal cell-derived factor-1
FLT-3 ligand= FMS-like tyrosine kinase 3 ligand
TNF-α = Tumour necrosis factor-alpha
TGF-β = Transforming growth factor beta
The cells of the immune system consist of lymphocytes, specialized cells that capture and
display microbial antigens, and effector cells that eliminate Microbes.
 MononuclearCells
Mononuclear Cells
• Monocytes in Blood, M in Tissues
– Monocytes 5-10 times smaller than M
• M Increases Phagocytic Ability
• Secretes cytokines and Produces Hydrolytic Enzymes
• Named Based on Tissue They Reside
– Alveolar (lungs), Kupffer (liver), Microglial (brain), Osteoclasts (bone)
• Activated By Phagocytosis, Cytokines (IFN secreted by helper T cells) & LPS from
bacteria.
• Antigen Presenting Capacity Thru MHC II
• Macrophages cause inflammation through the production of interleukin-1, interleukin-
6, and TNF-alpha
• They also have receptors for IgG & complement system
• There main role is to remove the antigens from the body. They are long lived months to
years
• Horse shoe shape nucleus and contain well developed golgi complexes and lysosomes
MONOCYTES AND MACROPHAGES!
Monocytes develop in the bone marrow and reach maturity in the blood. Mature
monocytes have large, smooth, lobed nuclei and abundant cytoplasm that contains
granules. Monocytes ingest foreign or dangerous substances and present antigens to
other cells of the immune system. Monocytes form two groups: a circulating group and a
marginal group that remain in other tissues (approximately 70% are in the marginal
group).
Most monocytes leave the blood stream after 20–40 hours to travel to tissues and
organs and in doing so transform into macrophages or dendritic cells depending on the
signals they receive. There are about 500 million monocytes in one litre of human blood.
M Effective APC
 Phagocytosis
Macrophages
Bifunctional
APC
Antigen
presentation

Liver
Skin
CNS lung
Kuppfer cells
Denteritic cells
Microglia
Alveolar
Bone – Osteovlast Connective tissue – Histiocyte
Placenta – Hoffbauer cells Spleen – Littoral cells
Kidney – Mesangial cells
 Dendritic cell
(APC)
• Long membrane extensions that resemble the dendrites of nerve cells.
• Dendritic cells (antigen presenting cell) can be difficult to isolate.
• It acts as a bridge between innate and adaptive immunity
• Dendritic cells express both MHC class I & class II molecules.
• capture antigen or bring it to the lymphoid organs where an immune
response is initiated.
 Developmental
Classification
A. By source Pathway of DCs
Myeloid DC
Lymphoid DC
B. By maturity
Immature DC
Mature DC
C. By distribution
Lymphoid tissues
Non-lymphoid tissues
Body fluid
DCs are found in many organs throughout the body
– DC in lymphoid tissue
• Interdigitating cell, IDC
The dendritic cells found in T cell areas of secondary lymphoid tissues are termed
interdigitating cells.
• Follicular DC, FDC
Follicular dendritic cells (FDC) are cells of the immune system found in primary and
secondary lymph follicles (lymph nodes) of the B cell areas of the lymphoid tissue.
• Thymic dendritic cell, TDC
Thymic dendritic cells (DC) delete self-antigen–specific thymocytes in the thymus
– DC not in lymphoid tissue
• Langerhans cells
Langerhans cells (LCs) are a specialized subset of dendritic cells (DCs) that
populate the epidermal layer of the skin.
• Interstitial DC
DC presents in the heart, lung, liver and intestine
– DC in body fluid
• Veiled cells
Antigen-presenting Cells of dendritic cell morphology found in the afferent LYMPH
and possessing veil-like processes.
• Peripheral blood DC
Dendritic cells activation. Extracellular signals, such as PAMPs or DAMPs, trigger
alterations on immature DCs culminating on significant changes on surface proteins,
intracellular pathways and metabolic activity
 Plasmacytoid dendritic cells (pDCs)
The plasmacytoid dendritic cell
(pDC) constitutes a unique DC subset
that links the innate and adaptive arm
of the immune system. Plasmacytoid
dendritic cells (pDCs) were originally
described in human lymph nodes in
the 1950s. pDCs specialize in
producing type I IFNs following their
recognition of viruses or self nucleic
acids through Toll-like receptor 7
(TLR7) and TLR9.

However, pDCs can also secrete

other pro-inflammatory cytokines and
chemokines, including interleukin-6
(IL-6), IL-12, CXC-chemokine
ligand 8 (CXCL8), CXCL10,
CC-chemokine ligand 3 (CCL3) and
CCL4. Moreover, expression of
MHC class II and co-stimulatory
molecules enables pDCs to present Plasmacytoid dendritic cells (pDCs) can be derived
antigens to CD4+ T cells. Thus, the from both myeloid and lymphoid progenitors.
biology of pDCs is multifaceted .
 Function of DC

1. Capturing and processing antigens

2. Presenting antigens
During the maturation of DC , its ability of Ag capture and
processing decreases while its ability of Ag presenting is given a rise.
 Different features of the three types of APC

▪Macrophages are good at presenting extracellular bacteria

▪DCs present bacterial and viral antigens
▪B cells can efficiently present soluble antigens
Interferon-sensitive response element (ISRE)
PD-L1 (Programmed death ligand) engagement
promotes apoptosis
IDO (indolamine 2,3-dioxygenase)-depletion
of tryptophan
Hepatocellular carcinoma (HCC)
Colorectal cancer (CRC)
Head and neck squamous cell
carcinomas (HNSCC)
Although, each cell is capable of producing type I IFNs, plasmacytoid dendritic cells (pDCs) possess a
unique ability to rapidly produce large amounts of them.

Deficiency in type I IFN production increases the risk of more severe viral infections and the
development of certain allergic reactions, and supports tumor resistance; nevertheless, its
overproduction promotes autoimmune reactions.

Therefore, the tight regulation of type I IFN responses of pDCs is essential to maintain an adequate level
of immune response without causing adverse effects.
 Granulocytes or Polymorphonuclear
(PMN) Leukocytes
A group of white blood cells is collectively referred to as granulocytes or
polymorphonuclear leukocytes (PMNs).

Granulocytes are composed of three cell types identified as neutrophils,

eosinophils and basophils, based on their staining characteristics with certain
dyes.

These cells are predominantly important in the removal of bacteria and

parasites from the body.

They are more short-lived than macrophages.

They engulf these foreign bodies and degrade them using their powerful
enzymes (lysozyme, peroxidases, defensins, etc).
 Granulocytic Cells
Neutrophils
• They are short lived 2 – 3 days.
• Released from bone marrow at a rate of 7 million per minute.
• Neutrophils comprise 95 % of circulating granulocytes.
• The fine granules stain poorly with acidic and basic dyes.
• They have characteristic multi lobed nucleus and are 10 – 20 um in diameter.
• The most numerous which constitute the majority of leukocytes in the blood i.e. about
60 – 70 %.
• There main role is phagocytosis .
• Lytic enzymes & bactericidal substances such as Peroxides, lysozyme and various
hydrolytic enzyme present in primary and sec. granules.
• Like monocytes they adhere to endothelial cell lining of the blood vessels and move
toward tissues by squeezing between the endothelial cells to leave the circulation. This
process is known as Diapedesis (The migration of blood cells (especially leucocytes)
through the intact walls of blood vessels into the surrounding tissue). This process is
promoted by chemo attractants such as chemokines (cytokines).
• Neutrophils are the 1st cells to arrive. A number of substances produced during an
inflammatory response recruit neutrophils to a site of inflammation.
• They are ferocious eaters and rapidly engulf invaders coated
with antibodies and complement, and damaged cells or cellular debris. Neutrophils do
not return to the blood; they turn into pus cells and die.
 Eosinophils

• Eosinophils have a bi lobed nucleus with many cytoplasmic granules that stain with acidic dye
e.g. eosin red
• They comprise 2 – 5 % of the blood leukocytes in a healthy non allergic person.
• The granules of eosinophils are membrane bounded organelles.
• Certain stimuli will cause eosinophils to degranulate.
• Degranulation involves the fusion of intracellular granules with the plasma membrane and the
release of granule content into the extra cellular environment, which become cytotoxic, when
they are released on the surface of parasites.
• Thus, eosinophils are mainly involved in reactions against parasitic infections. Eosinophils are
not phagocytic, but they intoxicate nematodes and other parasites and bacteria. The
cytotoxic substances are major basic protein, which kill helminths, eosinophil cationic
protein (an extremely efficient killer of parasites and potent neurotoxins) and eosinophil
peroxidase (kills bacteria, helminths and tumour cells). Eosinophils are involved in
hypersensitivity reactions.
• Eosinophils also release histaminases and aryl sulphatase which inactivate the mast cell
products e.g. histamine. The effect of eosinophils thus dampen down the effect of
inflammatory response.
 Basophils

• Circulating basophils and mast cells residing in the tissues are morphologically similar with
granules.
• Non phagocytic cell, lobed nucleus and stain with basic dye - Methylene blue.
• Basophils are found in a small number in the circulation, about le 0.01% to 0.3%.
• Basophils have membrane bounded granules.
• Basophils degranulate to release histamine, proteoglycans (eg Heparin & Chondroitin),
and proteolytic enzymes (e.g. elastase and lysophospholipase). They also
secrete lipid mediators like leukotrienes, and several cytokines. These histamine
containing cells are involved in hypersensitivity reactions.
• Play role in allergic responses when they release their granules (containing histamine,
serotonin, heparin, prostaglandin, etc into the bloodstream following exposure to specific
allergens).
• Basophils bear Fc receptors for IgE.
• When an individual is exposed to an allergen, allergen specific IgE is produced. This IgE
binds to the surface of basophils [in the sensitization phase of the allergic response]. Upon
re-exposure to the allergen, the allergen binds to IgE on the surface of basophils resulting in
degranulation [effector phase].
Cross-Linkage of Bound IgE Antibody With
Allergen Causes

Sensitization phase

Effector phase
 Mast cell
• Mast cells are very close to basophil in blood; the similarities between mast cells and basophils
have led many to speculate that mast cells are basophils that have "homed in" on tissues.
However, current evidence suggests that they are generated by different precursor cells in the
bone marrow. Nevertheless, both mast cells and basophils are thought to originate from bone
marrow precursors expressing the CD34 molecule.
• Basophils leave the bone marrow already mature, whereas the mast cell circulates in an
immature form, only maturing once in a tissue site. They don't differentiate until leaving
the blood vessels and enter the tissue.
• Mast cells are present in most tissues characteristically surrounding blood vessels and nerves,
and are especially prominent near the boundaries between the outside world and the internal
milieu, such as the skin, mucosa of the lungs and digestive tract, as well as in
the mouth, conjunctiva and nose.
• It contains cytoplasmic granules that secretes histamine.
• Development of allergy.
• The stimulus for these cells (Basophils and Mast cells) is an allergen – an antigen causing
allergic reaction.
• To be effective an allergen must cross link IgE molecules bounded to the surface of basophiles or
mast cells via its Fc receptors for IgE. Degranulation of mast cells or Basophils result in release
of all contents very rapidly.
Immune complexes bind to activating Fc receptors (FcR) and inhibitory FcRs that are expressed by innate immune effector cells such as
basophils, mast cells, neutrophils, monocytes and macrophages, in which they trigger the indicated effector responses. Binding of immune
complexes to FcRs on dendritic cells results in phagocytosis and presentation of antigenic peptides on MHC class I and class II molecules.
Antigen-specific CD8+ cytotoxic T cells, CD4+ helper T cells or regulatory T cells (TReg cells) that recognize these peptide–MHC complexes
become activated and mediate various effector functions such as killing of virus-infected cells, modulation of immune responses or providing
T-cell help for antigen-specific B cells. B cells only express the inhibitory low-affinity FcR for IgG (FcRIIB), which regulates activating signals
transduced by the B-cell receptor (BCR). On plasma cells, which produce high levels of antigen-specific antibodies, BCR expression is very low
or absent, resulting in exclusive triggering of inhibitory signalling pathways.
 Platelets

• Platelets, or thrombocytes, are small, irregularly shaped clear cell fragments (i.e.
cells that do not have a nucleus containing DNA), 2–3 µm in diameter, which are
derived from fragmentation of precursor megakaryocytes in the bone marrow.
• The average lifespan of a platelet is normally just 5 to 9 days. Platelets are a natural
source of growth factors. They circulate in the blood of mammals and are involved
in hemostasis, leading to the formation of blood clots.
• Platelets in addition to their role in blood clotting, are also involved in immune
responses, specially in inflammation.
• Platelets express Class I MHC products, receptors for IgG & IgE.
• In addition they also carry receptors for Factor VIII and other molecules for there
function.
 RBC - Erythrocytes

Red blood cells are the most common type of blood cell and the vertebrate organism's principal
means of delivering oxygen (O2) to the body tissues via the blood flow through the circulatory
system. They take up oxygen in the lungs or gills and release it while squeezing through the
body's capillaries.
These cells' cytoplasm is rich in haemoglobin, an iron-containing biomolecule that can bind
oxygen and is responsible for the blood's red color.
In humans, mature red blood cells are flexible biconcave disks that lack a cell nucleus and
most organelles. 2.4 million new erythrocytes are produced per second. The cells develop in
the bone marrow and circulate for about 100–120 days in the body before their components are
recycled by macrophages. Each circulation takes about 20 seconds. Approximately a quarter of
the cells in the human body are red blood cells.

Adult humans have roughly 2–3 × 1013 (20-30 trillion) red blood cells at any given time,
comprising approximately one quarter of the total human body cell number (women have about 4
to 5 million erythrocytes per microliter (cubic millimeter) of blood and men about 5 to 6
million; people living at high altitudes with low oxygen tension will have more). Red blood cells
are thus much more common than the other blood particles: there are about 4,000–11,000 white
blood cellsand about 150,000–400,000 platelets in each microliter of human blood.
 Cells of the Immune System
• Lymphoid Cells
– B-cells, T-cells and Null cells (NK cells)
– In peripheral circulation, 55-70% T cells, 15-30%
B cells and 3% Null cells or Third population cells
(TPC)
– 99% of lymph node
– Nucleus occupies almost entire cell
– 6 m diameter
Lymphoid Cells

20-40% of circulating WBC in blood

Adaptive Immunity
 LYMPHOCYTES
Lymphocytes are the only cells with specific receptors for antigens and are thus the key mediators of adaptive
immunity. Although all lymphocytes are morphologically similar and rather unremarkable in appearance, they are
extremely heterogeneous in lineage, function, and phenotype and are capable of complex biologic responses and
activities.

Small 6µm, contain a single nucleus, little visible cytoplasm around their nucleus. Responsible for the specific
immune response. Represent 20-40% of circulating WBC in blood.

T lymphocytes and B lymphocytes and natural killer cells. T and B lymphocytes are small, motile, nonphagocytic
cells which cannot be distinguished from each other morphologically.

Once stimulated with antigen enlarges 15µm into a blast cell. Lymphoblasts further differentiate into effector cells
or memory cells. [Plasma cells, T-helper cells, T-cytotoxic cells].

The memory cells are long-lived cells that reside in the Go phase of the cell cycle until activated by a secondary
encounter with antigen.

Different lineages or different maturational stages of lymphocytes can be distinguished by their expression of
membrane CD molecules (Cluster of Differentiation (CD).

T cells respond to antigens. Some of them (CD4+) secrete lymphokines which act on other cells involved in the
immune response. Others (CD8+, cytotoxic) are able to cause lysis of infected cells.

The major function of B lymphocytes is the production of antibodies in response to foreign proteins of bacteria,
viruses, and tumor cells.

Antibodies are specialized proteins that specifically recognize and bind to one particular protein that specifically
recognize and bind to one particular protein. Antibody production and binding to a foreign substance or antigen,
often is critical as a means of signaling other cells to engulf, kill or remove that substance from the body.
Our immune system is responsible for
helping to eliminate invaders (antigens)
such as infectious organisms. The key cells
in your immune system are lymphocytes
known as B cells and T cells, which
originate in your bone marrow. After T cells
further develop in your thymus, all of your
immune system cells gather in your lymph
nodes and spleen. Antigens (triangular
shapes above) are ingested (1), partially
digested (2) and then presented to helper T
cells by special cells called macrophages
(3). This process activates the helper T cell
to release hormones (lymphokines) that
help B cells develop (4). These hormones,
along with recognition of further antigens
(5), change the B cell into an antibody-
producing plasma cell (6). The antibodies (Y
shapes above) produced can be one of
several types (IgG, IgM, IgA, IgE and IgD)
(7). The antibody "fits" the antigen much
like a lock fits a key. The antigen is thus
rendered harmless. The helper T cells also
aid in development of cytotoxic T cells (8),
which can kill antigens directly; memory T
cells are produced (9) so that re-exposure
to the same antigen will provide a more
rapid and effective response (10).
Role of MHC molecules
MHC (major histocompatibility complex) molecules bind to antigen fragments produced
by infected cells.
This complex moves to the cell surface, "presenting" the antigen to T cell receptors on
cytotoxic T cells and helper T cells.
 What are Null Cells?
Null cells are also called as third population cells. Small fractions (2%) of the
lymphocytes circulating in the blood are neither T cells nor B cells.

A null cell is a large granular lymphocyte without surface markers (CD3) or

membrane-associated proteins from B lymphocytes or T lymphocytes (BCR or
TCR).

Unlike T and B cells, null cells lack immunologic memory and specificity.

Members of null cells are

NK cells,
Killer cells or antibody dependent cytotoxic cells (ADCC), and
Lymphokine activated killer(LAK)cells.
 NK Cells
• Natural killer cells are usually null cells with surface marker CD 16 which bind to the Fc
portion of the IgG, and thereby destroy it.

• NK-cells do immune surveillance, attack pathogens and abnormal cells (Tumor Cells and
Virally Infected Cells).

• NK cells have a role in the natural or innate immune response. These cells appear in a
larger proportion in young animals, and they diminish as the animal grows. This means
that the role of NK cells is related to natural or innate immune response mechanisms,
and not to those mediated by a specific antigenic stimulation (acquired immunity).
 Natural Killer cells (NK Cells)

Natural killer cells are bigger than normal lymphocytes, they have more
cytoplasm and dense granules! They are unspecific, NK cells will kill any cell that
has been infected with a virus! They will also destroy tumour cells...

Actually, there are no guns involved!! The NK releases "perforin" molecules onto the
target cell, which punches holes in its plasma membrane. The NK cell then releases
cytotoxins which will induce apoptosis (programmed cell death) in the target cell.
 Natural Killer Cells
Patrol the body and attack virus-infected body cells and cancer cells

Recognize cell surface markers on foreign cells

Destroy cells with foreign antigens

Rotation of the Golgi toward the target cell and production of perforins

Release of perforins by exocytosis

Interaction of perforins causing cell lysis+

IL-15 (interleukin -15) potentially promotes NK

cells development from NK precursors (NKPs)
and IL- 2 could maintain NK cell's survival
 KIR, KAR on NK- Natural killers
• NK cells – part of lymphocytic line

• Receptors recognising changes

on self cells molecules (after viral
infection)

• KAR – killer activation

receptors – recognise MICA,
MICB (stress molecules) on self
cells that activates NK cells to
kill self cell.

• KIR – killer inhibition receptors

– monitors MHC I molecules.
Tumor and virus infected cells
decrease number of MHC I, that
decrease the possibility of binding
to MHC I molecules and
decrease the inhibítion of killers
MICA & MICB {MHC class I polypeptide-related sequence A & B}
NK cells contain “Killer activating receptors" (KARs) and "Killer Inhibitory Receptors"
(KIRs). When an NK cell binds to a target cell without class I MHC molecules, the
activating receptors trigger the killing activity in NK cell. Killer inhibitory receptors (KIRs)
convey an inhibitory signal to the NK cells if they encounter class I MHC molecules on
target cell surface.

Since viruses often suppress the expression of class I MHC in host cells, the virus-infected
cells happen to be susceptible for NK cells killing activity. In the same way, cancer cells
showing reduced or no class I MHC expression are vulnerable to killing by NK cells.

Research information in this field has shown that IL-15 (interleukin -15) potentially
promotes NK cells development from NK precursors (NKPs) and IL- 2 could maintain NK
cell's survival and drive their development from committed precursors.

Hence IL- 2 and IL -15 are vital for NK cell generation. Absence of IL - 15 signaling cause
severe reduction in the number of NK cells.
 LAK Cells

Lymphokine-activated killer cell (also known as a LAK cell) is a white blood cell that
has been stimulated to kill tumour cells.

If lymphocytes are cultured in the presence of Interleukin 2, it results in the

development of effector cells which are cytotoxic to tumour cells.

LAK cells respond to these lymphokines, particularly IL-2, by lysing tumor cells that
were already known to be resistant to NK cell activity.

LAK cells are specific to tumor cells and to not display activity against normal cells.
 Killer Cells (K) or ADCC
They share many characters with both T and B cells. These cells possess Fc receptors for binding
with IgG antibodies and hence they are capable of killing cells coated with IgG antibodies.

ADCC involves the attachment of tumor-specific antibodies to tumor cells and the subsequent
destruction of the tumor cell by immunocompetent cells. Fc receptors on immunocompetent cells
recognize the Fc portion of antibodies adhering to surface tumor antigens
 Identifying Cell Using the CD
Nomenclature
• CD Cluster Of Differentiation
• Over 300 CD Markers
• T cells, CD4 or CD8 and CD3
• B cells, CD19
• NK cells, CD 16 & CD 56
• Monocytes/Macrophages CD14
• Dendritic Cells, CD1c (Human), CD11c (mouse)
 Cluster of Differentiation (CD)
The cluster of differentiation (CD) is a protocol used for the identification and investigation of cell surface
molecules present on white blood cells. Physiologically, CD molecules can act in numerous ways, often acting
as receptors or ligands (the molecule that activates a receptor) important to the cell. CD for humans is numbered up
to 350 most recently. Cell populations are usually defined using a '+' or a '-' symbol to indicate whether a certain cell
fraction expresses or lacks a CD molecule. For example, a "CD34+, CD31-" cell is one that expresses CD34, but not
CD31.

Type of cell CD markers

Stem cells CD34+, CD31-
All Leukocyte groups CD45+
Granulocyte CD45+,CD15+,CD24+,CD114+
Monocyte CD45+,CD14+,CD114+
T – Lymphocyte CD45+,CD3
T – Helper cell CD45+,CD3+,CD4+
Cytotoxic T cell CD45+,CD3+,CD8+
B – Lymphocyte CD45+,CD19+ or
CD45+,CD20+,CD24+
Thrombocyte CD45+,CD61+
Natural killer cell CD16+,CD56+,CD3-
 Suppressing the function of both
cytotoxic and Th cells. (suppress
excesive Immune reactions that
might be severely damaging the
body). Prevent the attack a
person’s own body tissues
(immune tolerance)

IL6
IL21

Tfh

CXCR5
CXCR5 – Chemokine
receptor type 5
The germinal center (GC) is a specialized microstructure that forms in secondary lymphoid
tissues, producing long-lived antibody secreting plasma cells and memory B cells, which can
provide protection against reinfection.
IL 6
IL 21

Follicular helper T cells (TFH): A subset of naive T cells in the T cell zone are activated by antigen and
migrate to the follicles where they differentiate into TFH cells which interact with and instruct
Follicular B (Fo B) cells to undergo isotype switching, somatic hypermutation, and rapid cellular
division to seed germinal centers (GC). Within these germinal centers, TFH cells continue to provide
help to GC B cells to facilitate their production of high affinity antibody producing plasma cells (PC)
and long-lived memory (Mem) B cells.
Naïve cells – Not exposed to antigen or virgin cells

Prime cells – Exposed to antigen first time

Reciprocal inhibition of Th1 and Th2 cells. Helper T Cells

IFN-γ (shown above in red) and IL-2 inhibit the

formation of Th2 cells;
IL-4 suppresses Th1 formation (shown above in
red).
 H1 activate B cells to produce
Antibody G & M (GM)

H2 activate B cells to produce

Antibody A, G & E (AGE)

Macrophage
binding antibody

Cytokines produced by TH1 and TH2 cells and their effects on the immune system. TH1 responses are initiated by IL-12 and interferon-IFN;,
and TH2 responses by IL-4. TH1 cells promote inflammation and production of complement and macrophage-binding antibody (solid blue
lines) and inhibit TH2 responses (dotted blue lines). TH2 cells promote humoral responses (solid red lines) and inhibit TH1 responses (dotted
red lines). Colored square denotes end result. ADCC, Antibody-dependent cellular cytotoxicity; APC, antigen-presenting cell; CTL, cytotoxic T
cell; DTH, delayed-type hypersensitivity; GM-CSF, granulocyte-macrophage colony-stimulating factor; TNF, tumor necrosis factor. & TGF-
tansforming growth factor.
H1 activate B cells to produce antibody G & M (GM)

H2 activate B cells to produce antibody A, G & E (AGE)

 MOLECULES OF THE INNATE MMUNE SYSTEM

A variety of molecules mediate protection against microbes during the period before
adaptive immunity develops. These molecules react with particular structures common to
a variety of microbes, and thus with many different microbes that express these structures.

Molecules of the innate immune system include

Complement
Acute phase proteins
Cytokines, particularly interferons
Anti-microbial peptides.
Some, especially those of the complement system, are vital for adaptive immunity.
 Major Cytokine-Producing Cells

• Innate (acute phase responses)

➢ Dendritic cells and macrophages: IL-1, TNF-α, TNF-β, IL-6, IL-12, GM-CSF,
chemokines, interferons α,β.
• Immune: T cells (CD4 and CD8)
➢ TH1 cells: IL-2, IL-3, GM-CSF, interferon-γ, TNF-α, TNF-β.
➢ TH2 cells: IL-4, IL-5, IL-6, IL-10, IL-3, IL-9, IL-13, GM-CSF, TNF-α.

Cytokines are small molecules that signal between cells, inducing growth, chemotaxis,
activation, enhanced cytotoxicity and/or regulation of immunity.

They are referred to as

Interleukins if produced primarily by leukocytes
Monokines if produced by myeloid cells,
Lymphokines if produced by lymphocytes, and
Chemokines if they direct cell migration.
Interferons protect against viral infection, inhibit protein synthesis, activate cells and modulate
immunity.
Lymphokines are growth factors for lymphocytes and influence the nature of the
immune response. IL-2 is made by T cells as a T cell growth factor. IL-3 is
important in hematopoiesis. IL-4 is produced by Th2 cells and mast cells and is a
growth and differentiation factor for Th2 cells and B cells. IL-5, also produced by
Th2 cells and mast cells, is important to B cell activation and production of IgA.
IL-10, which is produced by Th2 cells and MØ, induces Th2 responses.

Monokines have activities critical to immune defense and inflammation. IL-1,

tumor necrosis factor α (TNFα), and IL-6 activate lymphocytes, increase body
temperature, activate and mobilize phagocytes and activate vascular endothelium.
TNFα also activates MØ. IL-8 is chemotactic for PMNs. IL-12 activates NK cells
to produce IFNγ.
Other cytokines include colony-stimulating factors (CSFs) that drive development,
differentiation and expansion of cells of the myeloid series. GMCSF induces
commitment of progenitor cells to the monocyte/granulocyte lineage, G-CSF and
M-CSF commitment to the granulocyte or monocyte lineage, respectively.
Transforming growth factor β (TGFβ) inhibits activation of MØ and growth of B
and T cells. Tumor necrosis factor β (TNFβ) is cytotoxic.

Collectins, a group of carbohydrate-binding proteins, act as opsonins to facilitate

the removal and destruction of microbes. Peptide antibiotics, produced by a
variety of cells, are able to eradicate bacterial infections.
Interferon (IFN) proteins are a family of cytokines secreted by host cells to modulate the immune
response. As the first class of cytokines discovered, they were named “interferon” due to the protein’s
ability to interfere with viral replication. These signaling proteins are typically released by the host cell in
the presence of a pathogen and function to eradicate pathogens by alerting neighboring uninfected cells
to activate proper cell defense mechanisms. IFNs are classified into three types (Type I, Type II, and Type
III), according to the different receptors to which they bind (Table 1); each IFN type induces a specific
immune response. Additionally, IFN-mediated signaling promotes upregulation of major histocompatibility
class I and II molecules (MHC I, MHC II) and activates many downstream signaling cascades, resulting in
anti-viral defense machineries
Interferon types - Type I, II, III
Type I IFNs bind to a specific cell surface receptor known as IFN-α/β (IFNAR1, IFNAR2) and function as a
warning system for uninfected cells. In humans, Type I IFNs are the largest IFN family and include IFN-α,
IFN-β, IFN-ε, IFN-κ, and IFN-ω; they are produced by many cell types, including plasmacytoid dendritic
cells and fibroblasts. One major function of Type I IFN is the inactivation of eukaryotic translation
initiation factor 2a (eIF-2a), thereby inhibiting the synthesis of viral protein. Additionally, Type I IFN
activates RNase L, which cleaves any ssRNA within the cytoplasm, further inhibiting viral replication. IFN-
α has been used to treat hairy cell leukemia, while IFN-β has been used as treatment to slow the
progression of multiple sclerosis.

Type II IFNs (IFN-γ in humans) bind to the IFN-γ receptor complex (IFNGR1, IFNGR2) and are involved in
immune and inflammatory responses; they are produced by activated T cells and natural killer (NK)
cells. When Type II IFNs are released by T helper cells, type 1 (Th1 cells), leukocytes are recruited to the
sites of infection, leading to increased inflammation. Due to their role in the immune response,
unregulated Type II IFNs can lead to autoimmune diseases.

Type III IFNs include IFN-λ1, IFN-λ2, IFN-λ3, and IFN-λ4 and are implicated in the inhibition of viral
infections similar to Type I IFNs. IFN-λ1, IFN-λ2, and IFN-λ3 were originally named IL-29, IL-28a, and IL-
28b, respectively; IFN-λ4 is the newest Type III IFN to be discovered. Type III IFNs bind to the receptors
IFRL1 and IL-10R2, which are distinct from Type I receptors.
 Intracellular pathways induced by
Type 1 IFNs.

Type I IFNs bind to their respective

receptors and induce transcription of
PKR and OAS proteins. Once outside
of the nucleus, PKR and OAS proteins
contribute to an anti-viral state within
the cell. Upon activation, PKR
inactivates eIF-2α, therefore inhibiting
viral protein translation. OAS presence
in the cell contributes to activation of
RNase L, which functions to cleave
any viral ssRNA found in the cytosol.

Protein kinase R (PKR)

oligoadenylate synthetases (OAS)
This group of more than 50 small, closely related cytokines (MW 8–10 kDa)
are primarily involved in chemoattraction of lymphocytes, monocytes and
Neutrophils. Chemokines are small cytokines produced by many cell types in
response to infection or physical damage. They activate and direct effector
cells expressing appropriate chemokine receptors to sites of tissue damage and
regulate leukocyte migration into tissues. CC chemokines are chemotactic for
monocytes, CXC chemokines are chemotactic for PMNs.

One group has two adjacent cysteines (CC), a second has two cysteines
separated by another amino acid (CXC), another has one cysteine, and the last
has two cysteines separated by
three other amino acids.
 Acute Phase Proteins

Acute phase proteins are a heterogeneous group of plasma proteins important in innate defense against
microbes (mostly bacteria) and in limiting tissue damage caused by infection, trauma, malignancy and
other diseases.

They include

C-reactive protein (CRP) ,

Serum amyloid protein A (SAA), and
Mannose-binding protein (MBP).

Acute phase proteins are mainly produced in the liver, usually as the result of a microbial stimulus, or in
response to the cytokines IL-1, IL-6, TNFα and IFNγ that are released by activated macrophages and NK
cells.

These proteins maximize activation of the complement system and opsonization of invading microbes.
 B-lymphocytes

Produce specific antibodies or immunoglobulins to antigens. The immunoglobulins form part of the gamma-globulin
fraction in plasma. The B-lymphocytes multiply by clonal expansion. A specific antigen is bound and recognized to the B
cell through special surface immunoglobulins. B cells also contain CD19 and CD20 proteins. Cytokines activate the B-
lymphocyte, so it divides and the resulting cells differentiate to enormous plasma cells with an overwhelming surplus of
protein-producing endoplasmic reticulum (ER). This is why plasma cells produce large amounts of antibodies and release
them into the blood as Y-shaped molecules.

The plasma cells have a short life cycle, and die when they have fulfilled their defence mission. Hereby, the B-lymphocyte
population is reduced to its normal size apart from a few cells remaining as memory cells. The antibodies are also called
immunoglobulins (Ig). They are specific serum glycoproteins. Each antibody is Y-shaped and consists of heavy and light
polypeptide chains. The heavy chains with complement receptors provide the constant domain of the Ig molecule, which
is the same in all antibodies.

The light chain region constitutes the variable domain, which is functionally important. Antibodies deactivate antigens
by forming a complex, which causes agglutination and precipitation, by masking the active sites of the antigens, or by
activating the complement cascade. A single Ig with its antigen activates a complement cascade with mobilisation of up
to 109 new complement molecules carrying lots of enzymes that rapidly lyse the antigen-carrying microbe.
WHAT IS ANTIBODY?

.
Antibody is a protein used by the immune system
to identify and neutralize foreign objects like
bacteria and viruses each antibody recognize a
specific antigen unique to its target.
 Antibody Structure
• Antibodies are made up of:
– 2 light chains (identical) ~25 ,000 mol.wt
– 2 heavy chains (identical) ~50 ,000 mol.wt
• Each light chain bound to heavy chain by
disulfide (H-L)
• Heavy chain bound to heavy chain (H-H)
• First 110 a/a of amino terminal of both H and L
chain are variable, Hypervariable regions.
• Referred to as vL in light chains and vH in heavy.
• CDR (complementary determining regions)
• Remaining regions are very similar within same
class-constant regions.
Ig M&E (ME)-Lacks Hinge region
Isotypes
According to differences in their heavy chain
constant domains, immunoglobulins are grouped
into five classes, or isotypes: IgG, IgA, IgM, IgD,
and IgE.
 Antibody Structure
• Repeating Domains of ~110 a/a
– Intrachain disulfide bonds within each domain
• Heavy chains
– 1 VH and either 3 or 4 CH (CH1, CH2, CH3, CH4)
• Light chains
– 1 VL and 1 CL
• Hinge Region
– Rich in proline residues (flexible)
– Hinge found in IgG, IgA and IgD
– Proline residues are target for proteolytic digestion (papain and
pepsin)
– Rich in cysteine residues (disulfide bonds)
– IgM and IgE lack hinge region
– They instead have extra CH4 Domain
The most abundant is IgG, which has a high antigen affinity and is the antibody of the secondary response to
protein antigens (viruses and tetanus toxin). IgG can cross the placental barrier and protect the newborn for a
couple of months.

IgM is confined to the blood, because it is a pentameric molecule (5 IgM molecules joined together). IgM cannot
cross the placental barrier, and is responsible for the primary immune response.

IgA1 predominates in serum, whereas IgA1 and IgA2 are present in equal amounts in secretions such as saliva,
gastric juice, pancreatic and intestinal juice. IgA protects mucosal surfaces in the gut, respiratory and urinary
tracts, by preventing the attachment of poliovirus, enterovirus, bacteria, and enterotoxin.

The concentration of IgD in serum is high in disorders with B-lymphocyte activation such as AIDS and Hodgkin's
disease.

IgE is mainly bound to basophils and mast cells, and involved in the pathogenesis of allergic and nematode
diseases.
В lymphocytes are the only cells capable of producing antibodies; therefore, they are the cells that mediate
humoral immunity. В cells express membrane forms of antibodies that serve as the receptors that recognize
antigens and initiate the process of activation of the cells. Soluble antigens and antigens on the surface of
microbes and other cells may bind to these В lymphocyte antigen receptors and elicit humoral immune
responses. T lymphocytes are the cells of cell-mediated immunity. The antigen receptors of T lymphocytes
only recognize peptide fragments of protein antigens that are bound to specialized peptide display molecules
called major histocompatibility complex (MHC) molecules, on the surface of specialized cells called antigen-
presenting cells (APCs). Among T lymphocytes, CD4+ T cells are called helper T cells because they help В
lymphocytes to produce antibodies and help phagocytes to destroy ingested microbes. CD8* T lymphocytes
are called cytolytic, or cytotoxic, T lymphocytes (CTLs) because they kill cells harboring intracellular microbes,
that is, they lyse other cells. A third class of lymphocytes is called natural killer (NK) cells; these cells are
mediators of innate immunity and do not express the kinds of clonally distributed antigen receptors that В
cells and T cells do.
 Antibodies Act As Immunogens
• Antigenic Determinants on Abs Fall in 3
Categories Each immunoglobulin has unique
– Isotypic epitope(s) formed by
– Allotypic hypervariable region, probably
near or part of the antigen binding
– Idiotypic
site, called idiotope. These
• Isotypic antigenic determinants, together
– Constant Region Of Ab called idiotype.
– If you inject Ab in a different species Anti-Isotype
is generated
– If within same species, No Anti-isotype
 Antibodies Act As Immunogens
• Allotype
– Even though same isotypes within one species
small differences (1-4 a/a) arise in different
individuals (form of polymorphism)

– If injected with such Ab you generate anti-

allotype Ab
 Antibodies Act As Immunogens
• Idiotype
– Unique VH AND VL binds antigen but can also behave as
antigenic determinant
• If you inject a monoclonal antibody into a
genetically identical recipient then anti-idiotypic
antibodies are generated
• No anti-isotypic and no anti-allotypic Abs will be
generated
Comparison of T & B Cells
 Comparison of T & B Cells

Anamnestic response the rapid reappearance of antibody in the blood following introduction of an
antigen to which the subject had previously developed a primary immune response.
*Depending on subset.
CTL, Cytotoxic lymphocyte; DTH, delayed-type hypersensitivity; Ig, immunoglobulin; MHC, major histocompatibility complex; TCR,
T-cell receptor.
Surface markers of human B and T cells.