Magdy El Ekiaby, MD

Shabrawhi BTC
Host Defense
 Several mechanisms:
Natural barriers:
○ Skin and sweat
○ Mucous membranes and related secretions
Immune response:
○ Cellular
○ Humoral
Immune Response

Humoral Immunity Cell-mediated Immunity

(Antibody) (Cytotoxicity)
Antigen (Ag)
 A molecule which elicits a specific immune
response when introduced into an animal
 Antigenic (immunogenic) substances are
large molecules (>10,000 daltons in
molecular weight)
 Structurally complex (proteins are usually
very antigenic),
 Accessible (the immune system must be
able to contact the molecule)
 Foreign (not recognizable as "self").
Antibody (Ab)
 A glycoprotein produced in response to an
antigen that is specific for the antigen and
binds to it via non-covalent interactions
 The term "immunoglobulin" is often used
interchangeably with "antibody“
 The term "immunoglobulin" to describe any
antibody, regardless of specificity
 The term "antibody" to describe an antigen-
specific "immunoglobulin"
 Immunoglobulins (Igs) come in different forms
(IgA, IgD, IgE, IgG, IgM) that reflect their
structure
Structure of immunoglobulin
 Nomenclature of Igs:
 Isotype: Species specific
 Allotype: represents
genetic differences
between members of the
same species
 Idiotype; reflects the
antigen binding specificity
of any particular antibody
molecule. Idiotypes are so
unique that an individual
person is probably capable
of generating antibodies
directed against their own
idiotypic determinants
 GENERATION OF
ANTIBODY DIVERSITY
•The immune system has the capacity to
recognize and respond to about 107
different antigens. This extreme diversity
can be generated in at least three possible
ways:
•Multiple genes in the germ line DNA
•Variable recombination during the
differentiation of germ line cells into B-
cells.
•Mutation during the differentiation of
germ line cells into B-cells.
 Binds
Placental Binds mast cell Activates Additional
Isotype Structure phagocytic cell
transfer surfaces complement features
surfaces

First Ab in
IgM - - - + development
and response.

IgD - - - - B-cell receptor.

Involved in
opsonization and
IgG + - + + ADCC. Four
subclasses; IgG1,
IgG2, IgG3, IgG4.

Involved in
IgE - + - - allergic
responses.

Two subclasses;
IgA1, IgA2. Also
IgA - - - - found as dimer
(sIgA) in
secretions.
Kinetics of immune response
IMMUNOGLOBULIN
PRODUCTION
 The production of immunoglobulins by B-cells or plasma cells occurs
in different stages:
 During differentiation of the B-cells from precursor stem cells, rearrangement,
recombination and mutation of the immunoglobulin V, D, and J regions occurs to
produce functional VJ (light chain) and VDJ (heavy chain) genes.
 At this point, the antigen specificity of the mature B-cell has been determined. Each
cell can make only one heavy chain and one light chain, although the isotype of the
heavy chain may change
 Initially, a mature B-cell will produce primarily IgD (and some membrane IgM) that
will migrate to the cell surface to act as the antigen receptor.
 Upon stimulation by antigen, the B-cell will differentiate into a plasma cell expressing
large amounts of secreted IgM.
 Some cells will undergo a "class switch" during which a rearrangement of the DNA
will occur, placing the VDJ gene next to the genes encoding the IgG, IgE or IgA
constant regions
 Upon secondary induction (i.e. the secondary response), these B-cells will
differentiate into plasma cells expressing the new isotype. Most commonly, this
results in a switch from IgM (primary response) to IgG (secondary response).
 The factors that lead to production of IgE or IgA instead of IgG are not well
understood.
Cells of immune response
 Immune responsive cells can be divided
into five groups based on
presence of specific surface components
function
○ B-cells (B lymphocytes)
○ T-cells (T lymphocytes)
○ Accessory cells (Macrophages and other
antigen-presenting cells)
○ Killer cells (NK and K cells)
○ Mast cells
 Cell group Surface components Function
•Surface immunoglobulin (Ag
recognition) •Direct antigen recognition
•Immunoglobulin Fc receptor •Differentiation into antibody-
B-lymphocytes •Class II Major producing plasma cells
Histocompatability Complex •Antigen presentation within
(MHC) molecule (Ag Class II MHC
presentation)
•CD3 molecule
•Involved in both humoral and
T-lymphocytes •T-cell receptor (TCR, Ag
cell-mediated responses
recognition)

•Recognizes antigen presented

within Class II MHC
•Helper T-cells (TH) •CD4 molecule •Promotes differentiation of B-
cells and cytotoxic T-cells
•Activates macrophages

•Downregulates the activities of

•Suppressor T-cells (TS) •CD8 molecule
other cells
•Recognizes antigen presented
within Class I MHC
•Cytotoxic T-cells (CTL) •CD8 molecule
•Kills cells expressing
appropriate antigen

Accessory cells •Variable •Phagocytosis and cell killing

 Cell group Surface components
•Immunoglobulin Fc receptor
•Complement component C3b
•Macrophages
receptor
•Class II MHC molecule
•Dendritic cells •Class II MHC molecule
•Immunoglobulin Fc receptor
•Polymorphonuclear cells
•Complement component C3b
(PMNs) receptor
Function
•Bind Fc portion of
immunoglobulin (enhances
phagocytosis)
•Bind complement component
C3b (enhances phagocytosis)
•Antigen presentation within
Class II MHC
•Secrete IL-1 (macrokine)
promoting T-cell differentiation
and proliferation
•Can be "activated" by T-cell
lymphokines
•Antigen presentation within
Class II MHC
•Bind Fc portion of
immunoglobulin (enhances
phagocytosis)
•Bind complement component
C3b (enhances phagocytosis)
 Cell group Surface components Function

Killer cells •Variable •Direct cell killing

•Kills variety of target cells (e.g.

•NK cells •Unknown tumor cells, virus-infected cells,
transplanted cells)

•Bind Fc portion of
immunoglobulin
•K cells •Immunoglobulin Fc receptor •Kills antibody-coated target cells
(antibody-dependent cell-
mediated cytotoxicity, ADCC)

•Bind IgE and initiate allergic

Mast cells •High affinity IgE Fc receptors responses by release of
histamine
 Lymphoid tissue
Primary Secondary
(Responsible for maturation of Ag-reactive cells) (Sites for Ag contact and response)

Thymus
Bone marrow Lymph nodes Spleen
(T-cell maturation)

(Expansion of
lymphatic system,
(Similar to lymph
separate from blood
nodes but part of
circulation. Deep
(T-cell maturation) (B-cell maturation) blood circulation.
cortex harbors mostly
Collects blood-borne
T-cells, superficial
Ags)
cortex harbors mostly
B-cells)
Histocompatibility
 MAJOR HISTOCOMPATIBILITY COMPLEX
 The Major Histocompatibility Complex (MHC) is a set
of molecules displayed on cell surfaces that are
responsible for lymphocyte recognition and "antigen
presentation“
 The MHC molecules control the immune response
through recognition of "self" and "non-self" and,
consequently, serve as targets in transplantation
rejection
 The Class I and Class II MHC molecules belong to a
group of molecules known as the Immunoglobulin
Supergene Family, which includes immunoglobulins,
T-cell receptors, CD4, CD8, and others
MHC encoding genes
 The major histocompatibility complex is
encoded by several genes located on human
chromosome 6
 Class I molecules are encoded by the BCA
region
 Class II molecules are encoded by the D
region
 A region between these two on chromosome
6 encodes class III molecules, including some
complement components.
MHC I
 Class I molecules are composed of two
polypeptide chains; one encoded by the
BCA region and another (ß2-microglobulin)
that is encoded elsewhere
 The MHC-encoded polypeptide is about 350
amino acids long and glycosylated, giving a
total molecular weight of about 45 kDa
 This polypeptide folds into three separate
domains called alpha-1, alpha-2 and alpha-3
 ß2-microglobulin is a 12 kDa polypeptide
that is non-covalently associated with the
alpha-3 domain
 Between the alpha-1 and alpha-2 domains
lies a region capable of binding (via non-
covalent interactions) a small peptide of
about 10 amino acids. This small peptide is
"presented" to a T-cell and defines the
antigen "epitope" that the T-cell recognizes
MHC II
 Class II molecules are composed of
two polypeptide chains, both encoded
by the D region.
 These polypeptides (alpha and beta)
are about 230 and 240 amino acids
long, respectively, and are
glycosylated, giving molecular weights
of about 33 kDa and 28 kDa
 These polypeptides fold into two
separate domains; alpha-1 and alpha-2
for the alpha polypeptide, and beta-1
and beta-2 for the beta polypeptide
 Between the alpha-1 and beta-1
domains lies a region very similar to
that seen on the class I molecule. This
region is capable of binding (via non-
covalent interactions) a small peptide of
about 10 amino acids.
 This small peptide is "presented" to a T-
cell and defines the antigen "epitope"
that the T-cell recognizes
CLASS I vs CLASS II
MOLECULES
 Class I molecules are found on virtually every
cell in the human body
 Class II molecules, in contrast, are only found on
B-cells, macrophages and other "antigen-
presenting cells" (APCs).
 Class I molecules present antigen to cytotoxic T-
cells (CTLs) while class II molecules present
antigen to helper T-cells (TH-cells)
 This specificity reflects the third difference, the
type of antigen presented. Class I molecules
present "endogenous" antigen while class II
molecules present "exogenous" antigens
 An endogenous antigen might be fragments of
viral proteins or tumor proteins. Presentation of
such antigens would indicate internal cellular
alterations that if not contained could spread
throughout the body. Hence, destruction of these
cells by CTLs is advantageous to the body as a
whole.
 Exogenous antigens, in contrast, might be
fragments of bacterial cells or viruses that are
engulfed and processed by e.g. a macrophage
and then presented to helper T-cells. The TH-
cells, in turn, could activate B-cells to produce
antibody that would lead to the destruction of the
pathogen
T-CELL RECEPTOR (TCR)
MOLECULES
 The T-cell receptor molecule (TCR) is
structurally and functionally similar to the
B-cell immunoglobulin receptor
 TCR is composed of two, disulfide-linked
polypeptide chains, alpha and beta, each
having separate constant and variable
domains much like immunoglobulins
 The variable domain contains three
hypervariable regions that are
responsible for antigen recognition.
Genetic diversity is ensured in a manner
analogous to that for immunoglobulins
 TCR allows T-cells to recognize their
particular antigenic moiety
 However, T-cells cannot recognize
antigen without help; the antigenic
determinant must be presented by an
appropriate (i.e. self) MHC molecule
 Upon recognition of a specific antigen,
the signal is passed to the CD3 molecule
and then into the T-cell, prompting T-cell
activation and the release of lymphokines .
ANTIGEN RECOGNITION BY
T-CELLS
 The TCR provides the specificity for
an individual T-cell to recognize its
particular antigen
 However, this recognition is "MHC-
restricted" because the TCR also
requires interactions with MHC.
 Interactions betweenCD4 molecule
(found on helper T-cells), class II
MHC and consummate the antigen
recognition process, allow helper T-
cells to respond to "exogenous"
antigens (leading to B-cell
activation and the production of
antibody)
 CD8 molecule (found on cytotoxic
T-cells) and class I MHC stabilize
and consummate the antigen
recognition process, allowing
cytotoxic T-cells to respond to
"endogenous" antigens (leading to
target cell destruction)
Humoral Immunity
 The production of antibody involves three distinct
phases:
Induction phase: Ag reacts with specific T and B
cells
Expansion and Differentiation phase: Induced
lymphocyte clones proliferate and mature to a
functional stage (i.e. Ag receptor cells mature to
Ag effector cells)
Effector phase: Abs or T cells exert biological
effects either: independently or through the action
of macrophages, complement, other non-specific
agents
Regulation of humoral immune
respnse

Up-regulation Down regulation

Antigen-Antibody
Interactions
 Antigen/antibody affinity
 Antigen /antibody ratio
 Cross reactivity
Cell Mediated Immunity
 Cell mediated immune response is
effected by immune cells:
Phagocytosis and killing of intracellular
pathogens
Direct cell killing by cytotoxic T cells
Direct cell killing by NK and K cells
 These responses are especially
important for destroying intracellular
bacteria, eliminating viral infections and
destroying tumor cells
Macrophage activation
 Extracellular microorganisms:
 Non-encapsulated microorganisms are easily
phagocytosed and killed within macrophages
 Encapsulated microorganisms require the production of
antibody in order to be effectively phagocytosed.
 Intracellular microorganisms :
 Intracellular microorganisms elicit the production of
antibody, which allows effective phagocytosis. Once
engulfed, however, they survive within the phagocyte and
eventually kill it.
 Intracellular microorganisms also activate specific T-cells,
which then release lymphokines (e.g. IFN, TNF) that cause
macrophage activation. Activated ("killer") macrophages are
then very effective at destroying the intracellular pathogens.
CELL MEDIATED
CYTOTOXICITY
 The second half of the cell-mediated
immune response is involved in rejection
of foreign grafts and the elimination of
tumors and virus-infected cells.
 Effector cells:
cytotoxic T-lymphocytes (CTLs)
 NK-cells
 K-cells.
Cytotoxic T-lymphocytes
(CTLs)
 CTLs require recognition of a specific antigenic determinant and
recognition of "self" MHC
 CTLs recognize antigen via their T-cell receptor which makes specific
contacts with the antigenic determinant and the target cell's class I
MHC molecule.
 CTLs also express CD8, which may assist the antigen recognition
process.
 Once recognition is successful, the CTL "programs" the target cell for
self-destruction through
 CTLs may release a substance known as perforin in the space between the CTL and its
target which in the presence of calcium ions, the perforin polymerizes, forming channels
in the target cell's membrane. These channels may cause the target cell to lyse.
 CTL may also release various enzymes that pass through the polyperforin channels,
causing target cell damage.
 The CTL may release lymphokines and/or cytokines that interact with specific receptors
on the target cell surface, causing internal responses that lead to destruction of the
target cell
 CTLs principally act to eliminate endogenous antigens.
Killer cells
 NK cells are part of a group know as
the "large granular lymphocytes“
 These cells are generally non-
specific, MHC-unrestricted cells
involved primarily in the elimination
of neoplastic or tumor cells
 The precise mechanism by which
they recognize their target cells is
not clear
 Probably, there is some type of NK-
determinant expressed by the target
cells that is recognized by an NK-
receptor on the NK cell surface

Once the target cell is recognized,
killing occurs in a manner similar to
that produced by the CTL
 K-cells are probably not a
separate cell type but rather a
separate function of the NK group
 K-cells contain immunoglobulin
Fc receptors on their surface and
are involved in a process known
as Antibody-dependent Cell-
mediated Cytotoxicity (ADCC)
 ADCC occurs as a consequence
of antibody being bound to a
target cell surface via specific
antigenic determinants expressed
by the target cell
 Once bound, the Fc portion of the
immunoglobulin can be
recognized by the K-cell. Killing
then ensues by a mechanism
similar to that employed by CTLs
Complement
 The complement system found in the blood of mammals is
composed of heat labile proteins that combine with
antibodies or cell surfaces
 This complex, multicomponent system is composed of about
26 proteins
 The "complement cascade" is constitutive and non-specific
but it must be activated in order to function. The functions of
complement include:
 making bacteria more susceptible to phagocytosis
 directly lysing some bacteria and foreign cells
 producing chemotactic substances
 increasing vascular permeability
 causing smooth muscle contraction promoting and mast cell
degranulation
Complement Components
 Consists of 3 components:
Classical pathway
Alternative pathway
Membrane attack complex
Components of the Classical Pathway
Native component Active component(s) Function(s)

Binds to antibody that has

C1q
bound antigen, activates C1r.
C1(q,r,s) Cleaves C1s to activate
C1r
protease function.
C1s Cleaves C2 and C4.
C2a Unknown.
C2 Active enzyme of classical
C2b
pathway; cleaves C3 and C5.
Mediates inflammation;
C3a
anaphylatoxin.
Binds C5 for cleavage by
C3 C2b.
C3b Binds cell surfaces for
opsonization and activation
of alternate pathway.
C4a Mediates inflammation.

C4 Binds C2 for cleavage by

C4b C1s. Binds cell surfaces for
opsonization
Components of the Alternate Pathway
Native component Active component(s) Function(s)

Mediates inflammation;
C3a
anaphylatoxin.

C3
Binds cell surfaces for
C3b opsonization and activation
of alternate pathway.

Binds membrane bound C3b.

B
Cleaved by Factor D.

Factor B Ba Unknown.

Cleaved form stabilized by P

Bb
produces C3 convertase.

Cleaves Factor B when

Factor D D
bound to C3b.

Binds and stabilizes

Properdin P
membrane bound C3bBb.
Components of the Membrane-Attack Complex
Native component Active component(s) Function(s)

Mediates inflammation;
C5a
anaphylatoxin, chemotaxin.

C5
Initiates assembly of the
C5b membrane-attack complex
(MAC).

Binds C5b, forms acceptor

C6 C6
for C7.

Binds C5b6, inserts into

C7 C7 membrane, forms acceptor
for C8.

Binds C5b67, initiates C9

C8 C8
polymerization.

Polymerizes around C5b678

C9 C9n to form channel that causes
cell lysis.
Classical Pathway Alternative Pathway
REGULATION OF THE
COMPLEMENT CASCADE
 Because both the classical and alternate pathways depend
upon C3b, regulation of the complement cascade is mediated
via 3 proteins that affect the levels and activities of this
component.
 C1 Inhibitor inhibits the production of C3b by combining with
and inactivating C1r and C1s. This prevents formation of the
C3 convertase, C4b2b.
 Protein H inhibits the production of C3b by inhibiting the
binding of Factor B to membrane-bound C3b, thereby
preventing cleavage of B to Bb and production of the C3
convertase, C3bBb.
 Factor I inhibits the production of C3b by cleaving C3b into
C3c and C3d, which are inactive. Factor I only works on cell
membrane bound C3b, mostly on red blood cells (i.e. non-
activator surfaces).