Development of Immune

Responses (HIR and CMIR)

Dr.D.RATHNAMMA
Professor and Head
Dept. of Veterinary Microbiology,
Veterinary College, KVAFSU,
Hebbal, Bengaluru-560 024
Two types/arms of immune responses:
1. Humoral immune response: Effectors of the
humoral immunity are secreted antibodies, highly
specific molecules that can bind and neutralize
antigens on the surface of cells and in the
extracellular spaces. The primary domain of
antibody protection lies outside the cell.
2. Cell mediated immune (CMI) response:
• Principal role of CMI is to detect and eliminate
cells that harbour intracellular pathogens.
• Cell mediated immunity can also recognize and
eliminate cells, such as tumour cells, that have
undergone genetic modifications so that they
express antigens not typical of normal cells.
 Phases of Immune response
• Generation of specific immune response (HIR or
CMIR) that takes place after antigen stimulation
can be divided into three phases;
1. Recognition phase: lymphocytes recognize
antigen through their antigen specific receptors and
bind to antigen
2. Activation phase: Proliferation and differentiation
of antigen activated lymphocytes.
3. Effector phase: Elimination of antigen by effector
cells (Plasma cells in HIR and CTL in CMIR).
 T cell activation and differentiation

• Activation of a naive T cell in a secondary lymphoid organ

results in the generation of effector and memory T cells.
• T cell activation requires;
• Several receptor-ligand interactions between the T
cell and an APC.
• Signals through cytokines produced by the activating
APC and other supportive cells in the lymphoid organ.
• Effector CD4+ T cells become TH cells and secrete cytokines
that enhance the activity of many other immune cells.
• Effector CD8+ T cells become CTLs that kill infected cells.
 Costimulatory signals are required for optimal T-cell
activation and proliferation
Ag binding to PRRs on DCs and other APCs

Activated to express costimulatory ligands (CD80 & CD86)

CD28 is the costimulatory receptors on T cells

Produce cytokines that enhance their ability to activate T

cells
Positive costimulatory receptors and ligands
• Negative costimulatory receptors and ligands
– Inhibit TCR signaling
– Maintaining peripheral T-cell tolerance
– Reducing inflammation both after the natural
course of an infection and during responses to
chronic infection.
 Immune regulation

• Positive and negative co stimulation is carefully

regulated both temporally and spatially.
• Naïve T cells do not express negative co
stimulatory receptors
• Activated in secondary lymphoid tissue during
the initiation of an immune response.
• Effector T cells up-regulate negative co
stimulatory receptors at the end of an immune
response.
 Positive costimulatory receptors

• CD28
• 44 kDa glycoprotein, homodimer
• Expressed by all naïve and activated CD4+ T cells
and CD8 +T cells
• Enhances TCR-induced proliferation and survival
by cooperating with TCR signals to induce
expression of the pro-proliferative cytokine IL-2
and pro survival bcl-2 family member (bcl-xL).
• CD28 binds to
2 distinct ligands of the B7 family of proteins on APC
CD80 (B7-1)
CD86 (B7-2)
 Negative costimulatory receptors
• CTLA-4 (CD152)
– 2nd member of CD28 family
– Binding to both CD80 and CD86.
– It antagonise T-cell activating signals.
– Not expressed constitutively on resting T cells.
– Induced within 24 hrs after activation of a naïve T cell
– Peaks in expression within 2-3 days post-stimulation.
– Peak surface levels of CTLA-4 are lower than peak
CD28 levels.
– It binds to CD80 & CD86 with markedly higher affinity,
it competes very favourably with CD28.
• CTLA-4 expression levels increase in proportion to
the amount of CD28 costimulation.
• It acts to “put the brakes on” the proliferative
influence of TCR-CD28 engagement.
• Inhibitory function.
Clonal anergy
• Absence of a suitable costimulatory signal, T cell
will become unresponsive to subsequent
stimulation , known as ‘Anergy’
• Interaction between negative costimulatory
receptors and ligands can also induce anergy.
Development of Humoral Immune Response
ANTIBODY RESPONSE
B LYMPHOCYTES
B cells are effective antigen-presenting cells

• Following Ag binding, Ag is internalized and degraded,

transported to RER where MHC class II molecules and
antigen fragments combine.
• Antigen-MHC class II complexes are carried to the B cell
surface, where they are presented to T helper cells .
• This activates the TH cells that then provide co-
stimulation to the B cell and permit its full activation.
• Since all the antigen receptors on a single B cell are
identical, each B cell can bind only one antigen. This
makes them much more efficient antigen-presenting
cells than macrophages.
Development of Humoral Immune Response:
B cell activation, proliferation and differentiation
Sequence of events that must occur for a B cell to respond to antigen:
Not only must the B cell be stimulated by antigen,
but it must also receive costimulation from T helper cells and their cytokines.

Veterinary Immunology, I.Tizard

Humoral Immune response can be divided into
Primary and Secondary immune response
Primary immune response
• First exposure to a foreign antigen, a lag phase
occurs in which no antibody is produced, but
activation , proliferation and differentiation of
B cells will take place.
• Lag phase is longer, some times as along as
weeks or months.
• First antibody produced is IgM (small amounts
of IgG also produced)
• The amount of antibody produced is low.
Secondary immune response
• Second exposure to the same antigen results
in accelerated antibody response.
• Lag phase is very short (2-3 days) because of
involvement of memory B cells
• Major antibody produced is IgG
• Amount of antibody produced is very high and
tends to remain high for a longer time
• Principle behind booster injections in
immunization
 Differences between primary and secondary
immune response
PRIMARY RESPONSE SECONDARY RESPONSE

Responding B cell Naïve B cell Memory B cell

Lag period 7-10 days 2-3 days

Immunoglobulin Ig M IgG
class/isotype
Magnitude of antibody Varies depending on 100-1000 times higher
response antigen
Antigens Thymus dependent and Thymus dependent
thymus independent
Antibody affinity Lower Higher
 Immunoglobulin class switching
• Mature B cells carry IgM and IgD specific for particular
antigen on their surface, interact with antigen, produce
IgM antibody (IgD gets lost) in response to first
exposure to antigen.
• Gene rearrangements of heavy chain constant region
genes permits elaboration of antibodies of same
antigenic specificity, but of different immunoglobulin
classes.
• Antigenic specificity remains same for life time of B cell
and Plasma cell, because specificity is determined by
variable region genes (V, D and J genes of heavy and
light chains)
• Memory B cells express membrane immunoglobulin
(mIg) of IgG or IgA or IgE.
Development of Cell mediated Immune
Response
T LYMPHOCYTES
• Cell mediated immunity consists of both helper T cells
(TH1) and several types of cytotoxic cells.
• Effector cytotoxic cells arise from both the adaptive
and innate immune systems and, therefore, include
both antigen specific and non specific cytotoxic cells.
• Antigen non specific (innate immunity) cytotoxic cells
that contribute to the clearance of target cells include
NK cells and non lymphoid cell types such as
macrophages, neutrophils, and eosinophils.
• Antigen specific (adaptive immunity) cytotoxic cells
include CD8+ T lymphocytes (TC cells), as well as the
CD4+ NKT cells, which, although derived from the T-cell
lineage, displays some useful features of innate
immune cell type.
• Cytotoxic T lymphocytes (CTL), Natural killer T
(NKT) , and Natural killer (NK) effector cells all
induce cell death by triggering apoptosis in
their target cells.
• Cytotoxic cells eliminate cells infected with
intracellular pathogens (virus, bacteria,
protozoa), tumor cells and cells that have
been stressed by extreme temperatures or
trauma.
• Cytotoxic T lymphocytes, or CTLs, are
generated by immune activation of T cytotoxic
(TC or CD 8+T) cells. These effector cells have
lytic capability and are critical in the
recognition and elimination of altered self-
cells (e.g., virus-infected cells and tumour
cells).
• Naive TC cells are incapable of killing target
cells and are therefore referred to as CTL
precursors (CTL-Ps) to denote their
functionally immature state. CTL-P has to be
activated to differentiate into a functional CTL
with cytotoxic activity.
During development of CMIR, endogenous / intracellular
antigens are processed and presented along with MHC class I
molecules. Antigen+MHC class I recognized by Tc cells (CTL-Ps).
Generation of CTLs from CTL-Ps appears to require at
least three sequential signals;
• An antigen-specific signal 1
is transmitted by the TCR
complex upon recognition of
peptide-MHC class I
complex.
• A co-stimulatory signal
(signal 2) transmitted by the
interaction of CD28 of CTL-P
and B7 of the infected cell.
• Signal induced by the
interaction of IL-2 with the
high-affinity IL-2 receptor,
resulting in proliferation and
differentiation of the
antigen-activated CTL-P into
effector CTLs
Generation of effector CTLs
Upon interaction with antigen+ MHC class I complexes on
appropriate target cells, CTL-Ps begin to express IL-2 receptors
(IL-2R) . Proliferation and differentiation of antigen-activated
CTL-Ps generally require IL-2 secreted by TH1 cells resulting from
antigen activation and proliferation of CD4+ T cells. In the
subsequent effector phase, CTLs destroy specific target cells.

Cytokines from TH1 cells

also regulate activation of
non specific effector
cells; NK cells and
Macrophages
 CTL mediated killing of target cell:
Primary events in CTL-mediated killing; conjugate formation,
membrane attack, CTL dissociation, and target cell destruction.
1. Directional delivery of cytotoxic proteins (perforin, a pore forming
protein and granzymes/ fragmentins, serine proteases) that are
released from CTLs and enter target cells.
2. Interaction of the membrane-bound Fas ligand on CTLs (death ligand)
with the Fas receptor on the surface of target cells.
• Either of these initiating events results in the activation of a signalling
pathway that culminates in the death of the target cells by apoptosis.

Within 5 min. of CTL contact,

target cell begins to exhibit
DNA fragmentation.
Cytotoxic T cells kill their targets through two pathways:

1. Perforin pathway
• Secretion of perforins and granzymes from secretory
lysosomes.

2. Fas (CD95, death receptor) pathway

• Signalling through the CD95 death receptor.
Perforin pathway

The killing process can be

divided into 3 phases:
1. Adhesion
2. Lethal hit
3. Cell death
 Fas (CD95) Pathway (Death receptor pathway)

• The binding of a T cell surface protein called CD95L

(Fas ligand or CD178) to a target cell death receptor
called CD95 (Fas).
• CD95L is expressed on activated CD8+ T cells and NK
cells. It binds to CD95 on target cells.
Fas (CD95) pathway of T cell mediated cytotoxicity
Destruction of target cells by NK cells:
• Natural killer cells appear to kill tumor cells and virus
infected cells by processes similar to CTLs.
• NK cells express FasL on their surface and readily
induce death in Fas-bearing target cells.
• Cytoplasm of NK cells contains numerous granules
containing cytotoxic proteins perforin and granzymes.
Unlike CTLs, which need to be activated before
granules appear(CTL-Ps lack cytoplasmic granules and
perforin; upon activation, cytoplasmic granules appear,
MHC restricted),
• NK cells are constitutively cytotoxic, always having
large granules in their cytoplasm. Release the granule
contents, perforin causes pores on the membrane,
granzymes induces death of target cells.
 NK cells mediate Antibody dependent cell mediated
cytotoxicity (ADCC)
Among the cells that can mediate
ADCC are NK cells, macrophages,
monocytes, neutrophils, and
eosinophils, express Fc receptors.
When antibody is specifically bound
to a target cell, Fc receptor-bearing
cells can bind to the Fc region of IgG
and thus to the target cells, and
subsequently cause lysis of the target
cell.
Although these cytotoxic cells are
non specific for antigen, the
specificity of the antibody directs
them to specific target cells.
• Target cell killing by ADCC appears to involve a number of
different cytotoxic mechanisms, but not complement
mediated lysis.
• When macrophages, neutrophils, or eosinophils bind to a
target cell by way of the Fc receptor, they become more
active metabolically; as a result, the lytic enzymes in their
cytoplasmic lysosomes or granules increase.
• Release of these lytic enzymes at the site of the Fc mediated
contact may result in damage to the target cell.
• In addition, activated monocytes, macrophages, and NK cells
have been shown to secrete tumor necrosis factor (TNF),
which may have a cytotoxic effect on the bound target cell.
• NK cells and eosinophils contain perforin in cytoplasmic
granules, their target-cell killing also may involve perforin-
mediated membrane damage similar to the mechanism by
CTL-mediated cytotoxicity.
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