IS LEC MIDTERM REVIEWER

CELL-MEDIATED IMMUNITY Cells that present antigens to T lymphocytes

and activate
Duality of the Immune System
 Express both Class I and Class II MHC
Cell Mediated Immunity molecules on surface.
• Provided by T lymphocytes  Also express B7 (B7.1 AND B7.2)
• Provides immunity to: Molecules
 Present antigens by Class I MHC molecules
(i) intracellular bacteria to CD8+A T cells
 Professional APCs are dendritic cells,
(ii) viruses,
Macrophages and B lymphocytes.
(iii) fungi,  APCs process and present antigens in two
pathways
(iv) protozoa and 1. Exogenous or Endocytic Pathway:
Phagocytosed microorganisms are degraded
(v) tumours
in the phagosomes and peptides are
• cells can recognize antigen only when it is presented in this pathway by Class II MHC
presented on the surface of Antigen presenting cells molecules
(APCs) by self MHC molecules 2. Endogenous or Cystosolic Pathway:
Intracellular microorganisms synthesize
• This self MHC restriction results from positive protein in the cytoplasm that are presented
selection during maturation of T cells in the thymus in this pathway by Class I MHC molecules.
• Involves specialized set of lymphocytes called T T lymphocytes
cells that recognize foreign antigens on the surface
of cells, organisms, or tissues: Helper T cells Maturation
Cytotoxic T cells
 Derived from bone marrow by
• T cells regulate proliferation and activity of other hematopoiesis
cells of the immune system: B cells, macrophages,  Progenitor T (Pro-T) cells migrate to thymus
neutrophils, etc.  Maturation occurs in the thymus
-Rearrange two selection process
• Defense against:
-Undergo two selection
 Bacteria and viruses that are inside host cells  Positive selection: Self MHC restriction
and are inaccessible to antibodies.  Negative selection :Self tolerance
 Fungi, protozoa, and helminthes  Acquire surface CD molecules e.g CD3,
 Cancer cells CD4/ CD8
 Transplanted tissue  Two population of T cells are released to the
circulation:
Antigen Presenting Cells
-CD4+ or helper T (TH) cells
 Dendritic Cell -CD8+ or Cytotoxic (Tc) cells
 Macrophage
Mechanism of CMI
 Lymphocyte
Antigen recognition by T cells
  -Antigen recognition by TCR provides
Stimulatory signal Signal 1 to the T cell
 -Binding of B7 molecule on APC with
CD28 molecule on T cell provides Co-
stimulatory signal (Signal 2) to the T cell.
 -T cells recognize specific antigens
presented with MHC molecules on the
surface of APCs by the TCR
 -Each T cell has 10^5 TCRs on its surface
all of which recognize a one antigen (or
epitope)
 -10^10 clones of T cells will recognize
10^10 antigens
T cell activation
-When T cells receive both Stimulatory (Signal 1)
and Co-stimulatorty (Signal 2) signals they are
activated (clonal activation)
-If the T cells receive only the Stimulatory signal
without Co-stimulatory signal they are permanently
inacticated (clonal energy)
T cell differentiation
-Activated T cells start to proliferate, synthesize and
secrete IL-2 and express IL- 2 receptors on cell
surface
-After several divisions they differentiate to effector
and memory T cell populations
-Memory T cells have long life span (20-30 years)
and provide immunity if the person is re-exposed to
the same antigen.
Effector T cells are short-lived (few days to weeks)
cells and carry out specialized functions e.g.
-CD8+ effector T cells: Induce apoptosis of virus
infected and tumour cell (Cytotoxic killing)
-CD4+ effector T cells: Secrete cytokines that cause
macrophage activation to kill intracellular
pathogens and help T cell and B cell activation.
Cytotoxic T cells induce apoptosis of infected
cells bearing antigen on the surface.
Antigen Elimination by CMI
CD4+ Effector T cells
-With influence of cytokines e.g . IL- 12 from
APCs, Th cells differentiate to TH1 cells
-TH1 cells release cytokines e.g interferon-y (IFN-
y)
-IFN-y activates macrophages that phagocytose and
eliminate intracellular pathogens.
Activated macrophages have more phagocytic
and killing activity and they phagocytose and kill
intracellular pathogens effectively.
Regulatory Role of Th cells
TH cells help Tc cell activation
Naïve cytotoxic T cells (cytotocic T cell
precursor) require cytokines from TH cells for
activation.
TH help B cells
 Activation of naïve B cells require
-Direct interaction with TH cells by
o Antigen in Clas II MHC and TCR
o CD40 and CD40L
o B7 and CD28
 Cytokine from Th1 cells also help T cell
activation
-Thus Th1 cells regulate CMI
-With the influence of IL-4 (from mast cells) Th
cells differentiate to TH2 cells
-Cytokines from TH2 cells help cell activation
-Thus TH2 cells regulate humoral immunity
Interaction between components of the immune
system
1. Internalized antigen digested by cell
2. Altered self-cell presents antigen
3. T cell receptors recognize antigen bound to toxin of S.aureus syndrome
MHC molecules Erythrogenic toxin of Scarlet Fever
4. Binding antigen-MHC activated cells S.pyogenes
5. Activated Th cell secretes cytokine that Pyrogenic toxin of Streptococcal Toxic
Streptococcus pyogenes Shock Syndrome
contribute to activation of B cells, Tc cells
Mycoplasma arthritidis Arthritis, Schock
and other cells (macrophages) supernatant
6. Activated CTIs recognize and kill altered
self-cells
7. B-cells interact with antigen and CYTOKINES
differentiate into antibody-secreting plasma
-Protein molecules secreted by cells that
cells
regulate function of that cell or other cells
8. Antibody binds antigen and facilities its
clearance from the body -The name denotes their role in cell to cell
communication
CD4+ T(Th) cells regulate the function of
both Tc and B cells -Development of an effective immune response
involves lymphocytes and other leukocytes
Th cell activation by Superantigen
-Cytokines play key role in the complex
Superantigens are viral or bacterial proteins
interaction between cells of Immune system
that bind simultaneously to the Vb domain of a
TCR and to the A chain of class II MHC Autocine: Acts on the same cell that secretes
molecule it
They bind outside of the TCR antigen binding  Paracrine : Acts on adjacent cells
cleft
Endocrine : Carried by the blood or bloody
Any T cell expressing a particular Vb fluid to a distant site and acts on distant cells.
sequence will be activated by a corresponding
superantigen -One cytokine may act on various cells and
produce various effects

-Many cytokines may act on the same cell and

Th cell activation by Superantigens produce same effect
Superantigen -One cytokine may increase action of the other
Enterotoxin of Staphylococcus aureus -One cytokine may inhibit action of the others

Superantigen -Cytokines secreted by some leukocytes and

acting on other leukocytes are called
Enterotoxin of Food Poisoning INTERLEUKINS
Staphylococcus
aureus -Cytokine secreted by lymphocytes are called
LYMPHOKINES
Toxic shock syndrome Toxic shock
toxin (TSS1) of syndrome -Cytokine secreted by monocytes and
S.aureus macrophafes are sometime called
MONOKINES
Exfoliative dermatitis Scalded skin
-Cytokines that cause chemotaxis of leukocytes
are called CHEMOKINES
Relationship Between CellMediated and
Humoral Immunity
Antibody Production
T-Dependent Antigens:
• Antibody production requires assistance from
T helper cells.
• A macrophage cells ingest antigen and
presents it to T H cell.
• TH cell stimulates B cells specific for antigen
to become plasma cells.
• Antigens are mainly proteins on viruses,
bacteria, foreign red blood cells, and hapten-
carrier molecules.
T-Independent Antigens:
• Antibody production does not require
assistance from T cells.
• Antigens are mainly polysaccharides or
lipopolysaccharides with repeating subunits
(bacterial capsules).
• Weaker immune response than for T-
dependent antigens.
CELL MEDIATED IMMUNE RESPONSES
• Primary Function Of Cell Mediated Response
• Eliminate intracellular pathogens
• Eliminate tumor cells
• Both Ag Specific and Non-specific cells Are
Involved
• Ag Specific: CD8+ Cells (TC) And TH (DTH)
• Non-specific: Macrophages, Neutrophils, NK
• Both Specific And Non-specific Require
Cytokines
• Humoral and Cell Mediated do collaborate
• Ex. Macrophages use Abs as receptors to
recognize target cells
Major histocompatibility complex
Introduction
Major Histocompatibility complex (MHC)
• is set of surface proteins located on the cell
membrane of nucleated cells.
• It plays more important work to identify the
antigen between self and non self body,
intracellular recognition and responsible for
antigen presentation.
• Histo refers to tissues. Compatibility refers to
living together harmoniously.
• MHC molecules always recognize only T
lymphocytes. The two types of MHC are
worked in immunity.
• T helper (Th) cell recognized by MHC
molecules II,
• T cytotoxic (Tc) cells are recognized by MHC
I molecules.
Major Histocompatibility complex (MHC) is
membrane attached protein which work on
recognition of antigen between self and non self
body and antigen presentation.
History
• Peter Gorer (1930) found that four group of
MHC molecules he used the blood sample of
mice to identified the blood group antigen
which designated by I to IV group of MHC.
• Georg Snell, Jean Dausset and Bariy received
noble prize in 1980 for their contribution to the
discovery of MHC molecule.
Aspects of MHC
1. MHC molecules are membrane-bound. Classes of MHC Molecules
Recognition by T cells requires cell-cell contact.
The MHC molecules are classified in to three
2. Peptide from cytosol associates with class I classes namely:
MHC and is recognized by Tc cells. Peptide
from vesicles associates with class II MHC and 1. Class I MHC molecules
is recognized by Th cells. 2. Class II MHC molecules
3. Although there is a high degree of 3. Class III MHC molecules
polymorphism for species, an individual has
maximum of six different class I MHC products Class I MHC Molecules
and only slightly more class II MHC products.
• Class I MHC(45 KD) molecule are a group of
4. A peptide must associate with a given MHC major histocompatibility antigen.
of that individual, otherwise no immune
• They are present on the surface of all nucleated
response can occur. That is one level of control.
cells except nervous tissue and platelets.
Mature T cells must have a T cell receptor that
recognizes the peptide associated with MHC. • It present antigen to Tc cells.
This is the second level of control.
• It bind with CD-8 adhesion molecules of Tc cells.
5. Each MHC molecule has only one binding • It brings about cell mediated immune response.
site. The different peptides a given MHC
molecule can bind all bind to the same site, but Structure of Class I MHC Molecule
only one at a time.
• It consists two polypeptide chains namely α chain
6. MHC polymorphism is determined only in and β2 – micro globulin.
the germ line. There are no recombination
• α chain which is non covalently attached with β2
mechanisms for generating diversity.
microglobulin . α chain contain a transmembrane
7. Because each MHC molecule can bind many glycoprotein which is encoded by A,B and C gene
different peptides, binding is termed degenerate. of grouped HLA.
8. Cytokines (especially interferon-γ) increase
• α chain is organized by three domains such as α1,
level of expression of MHC.
α2 and α3 each domain containing 90 amino acids
9. Alleles for MHC genes are co-dominant. sequences .
Each MHC gene product is expressed on the cell
• β2 microglobulin is similar in size of α 3 and it
surface of an individual nucleated cell.
dose not contain trans membrane proteins .
10.The control of transplantation,
• When the antigen is internalized and processed
autoimmunity, and the other immune responses
inside by proteosome (Ubiquitin, cytosolic
are the phenotypic consequences of the function
degradation), the peptides are produced .
of molecules encoded in the MHC.
• Peptide is further loaded on the groove of MHC I
MHC follows
molecules from endoplasmic reticulum.
 Polymorphism
Class II MHC Molecules
 Polygenism
 linkage disequilibrium
 Co- Dominance
• Class II MHC molecule are present on the surface
of antigen presenting cell and cell which engulfed
the foreign antigen.
• It binds with the exogenous(endocytic
degradation) antigens.
• It binds with CD4 adhesion molecules TH cells. •
It also consist of two polypeptide chains namely α
chain and β chain.
• Antigen is processed inside the endosome and
peptide is further loaded on groove of MHC II
molecules.
Structure of Class II MHC Molecule
• The class II MHC Molecule consists of two
polypeptide chain namely α chain (33 kDa) and β
(28kDa) chain.
• The both chain are attached noncovelantly.
• Each chain contains two units. The two units of α
chain are called α1 and α2. The two domains of β
chains are called β1 and β2.
• β2 and α2 are transmembrane domains anchoring
the MHC to plasma membrane.
• The α1 and β1 domains jointly bear a peptide
binding groove.
Class III MHC Molecules
• Class III proteins are secreted proteins that have
an immune function, but they are not expressed on
cell surfaces
• code for complement proteins and cytokines such
as tumor necrosis factor
HLA - Human Leukocyte Antigen
• HLA is the MHC molecules present in human
beings.
• HLA is a set of surface protein present on the
surface of all nucleated cells.
• They are responsible for graft rejection, adaptive
immunity, defense against infection, some time it is
expressed on cancer cell destruction, certain
autoimmune diseases and certain complements.
Note: MHC is the general term referring to the cell
surface antigen of vertebrates
H-2 Complex Of Mouse
• The major histocompatibility complex (MHC) of
mouse
• H-2 complex is a cluster of genes responsible for
the production of antigens located of nucleated cells
and complement components.
• This complex is located in the short arm of the
chromosome number 17.
• It consists of a set of structural genes .
• The genes, that make up a given
histocompatibility complex, are called halotypes.
Function of MHC Molecules
• MHC molecules are loaded with a bit of sample
peptide fragment derived from the degradation of
proteins present inside the cell. This peptide is the
mirror image of proteins present inside the cell. •
MHC molecules contain self as well as nonself
(foreign) antigen.
• They bring about defense against infections and
diseases.
• They mediate certain autoimmune diseases.
• They are responsible for individual smell of
people.
The complement system
HISTORY
• Research on complement began in the 1890s,
when Jules Bordet at the Institute Pasteur in
Paris showed that sheep antiserum to the
bacterium Vibrio cholerae caused lysis of the
bacteria and that heating the antiserum
destroyed its bacteriolytic activity.
• He named those substances as Alexins.
• Paul Ehrlich coined the term complement.
INTRODUCTION
• It is named “complement system” because it
was first identified as a heat-labile component
of serum that “complemented or augment”
antibodies in the killing of bacteria.
• Consists of serum and cell surface proteins
involved in defense against pathogens and tissue
damage mediated by antibodies
• The Complement system is the major effector
of cellular and humoral branch of immune
system.
• Plays major role in both innate and adaptive
immunity.
• Complement system represents a group of
about 20 proteins which augment or
complement the immune response.
• Most of these proteins are found in serum or
on cell surfaces.
• Synthesized in liver as inactive precursors and
are activated by proteolysis during their
interaction in a sequential manner.
• Also produced by blood monocytes, tissue
macrophages and epithelial cells of he
gastrointestinal and genitourinary tract.
General properties
• Present in serum of all animals but its
concentration is maximum in serum of guinea
pig.
• Complement of one species are able to react
with antibodies of other species but not to the
same extent.
• C- proteins constitute about 5% of normal
serum protein .
• Are glycoproteins.
• Are synthesized rapidly in inflammatory
responses – hence are called acute phase
proteins.
• Heat labile and lost activity at 56⁰ C for 30
mins and inactivated.
• Immunoglobulins are not inactivated at this
temperature.
• Binds with Fc potion of immunoglobulns .
Three main effects of complement
1. Lysis of cells (bacteria, allografts, tumor
cells)
2. Generation of mediators of inflammation
3. Opsonization – enhancement of phagocytosis
Complement System
• Complement proteins: are proenzymes -
activation by cleavage.
Complement Pathway
Three pathway of complement activation
1. Classical pathway:- Is antibody dependent
pathway and triggered by formation of soluble
antigen-antibody complex or by binding of the
antibody to the antigen present on the target cell
surface.
2. Alternative pathway:- Is antibody
independent pathway stimulated by antigen
directly eg. Bacterial cell surface components.
3. Lectin Pathway:- Also antibody independent
but resembles classical pathway.
Stages of complement activation
• Three main stages in the activation of
complement by any pathway are
 Formation of C3 convertase Definition of terms
 Formation C5 convertase
 Formation of membrane attack complex • C-activation: alteration of C proteins such
(MAC) that they interact with the next component

• The initiation and formation of C3 • C-fixation: utilization of C by Ag-Ab

convertase are different in classical and
alternative pathway . These then follow the
parallel route to merge at C5 convertase
stage and finally generate the MAC by a
common route.
• Sequential activation of complement
components occurs via one of three
pathways:
complexes
• C-inactivation: denaturation (usually by
heat) of an early C-component resulting in
loss of hemolytic activity
• Convertase/esterase: altered C-protein
which acts as a proteolytic enzyme for
another C-component.

 the classic pathway,

 the lectin pathway, and CLASSICAL PATHWAY
 the alternative pathway .
Native Active Functions
• Of these pathways, the lectin and the Component Components
alternative pathways are more important the C1(q,r,s) C1q Binds to
antibody that
first time we are infected by a
has bound
microorganism because the antibody antigen,
required to trigger the classic pathway is not activates C1r
present. C1r Cleaves C1s to
activate
• The lectin pathway and the alternative protease
pathway are, therefore, participants in the function.
innate arm of the immune system. C1s Cleaves C2 and
C4
• All three pathways lead to the production C2 C2a Unknown
of C3b, the central molecule of the C2b Active enzyme
complement cascade. of classical
pathway;
• The presence of C3b on the surface of a cleaves C3 and
microbe marks it as foreign and targets it for C5.
destruction. C3 C3a Mediates
inflammation;
• C3b has two important functions: anaphylatoxin
C3b Binds C5 for
(1) It combines with other complement cleavage by
components to generate C5 convertase, the C2b. Binds cell
enzyme that leads to the production of the surfaces for
opsonization
membrane attack complex and
and activation
(2) It opsonizes bacteria because phagocytes of alternate
pathway.
have receptors for C3b on their surface.
C4 C4a Mediates
 inflammation • Other activators include:
C4b Binds C2 for
cleavage by 1. Complexes containing IgA
C1s. Binds cell
surfaces for 2. Some virus-infected cells (e.g. EBV)
opsonization.
3. Many gram negative and gram positive
C5 C5a Mediates organisms
inflammation;
anaphylatoxin, 4. Parasites – Trypanosomes, Leishmania
chemotaxin.
C5b Initates 5. Erythrocytes
assembly of the
membrane- 6. Carbohydrates (agarose)
attack complex
(MAC) ALTERNATIVE PATHWAY
C6 C6 Binds C5b,
forms acceptor Native Active Functions
for C7. Component Components
C7 C7 Binds C5b6, C3 C3a Mediates
insets into inflammation;
membrane, anaphylatoxin
forms acceptor C3b Binds cell
for C8 surface for
C8 C8 Binds C5b67, opsonizaiton
initiates C9 and activation
polymerization. of alternate
C9 C9n Polymerizes pathway.
around C5b678 Factor B B Binds
to form membrane
channel that bound C3b.
causes cell Cleaved by
lysis. Factor D
Ba Unknown
Bb Cleaved form
Alternative pathway stabilized by
P produces
• Ab independnt pathway. C3
convertase.
• In the alternative pathway, many unrelated Factor D D Cleaves
cell surface substances, e.g., bacterial Factor B
lipopolysaccharides (endotoxin), fungal cell when bound
walls, and viral envelopes, can initiate the to C3b
Properdin P Binds and
process by binding C3 and factor B.
stabilizes
• This complex is cleaved by a protease, membrane
bound C3bBb
factor D, to produce C3bBb - acts as a C3
convertase to generate more C3b.
Lectin pathway
• Usually activated by products of micro-
organisms like endotoxin • Also known as the MBL Pathway
• In the lectin pathway, mannan-binding
lectin (MBL) (also known as mannose-
binding protein) binds to the surface of
microbes bearing mannan (a polymer of
the sugar, mannose).
• Binding causes activation of MASP
(MBP- associated serine proteases)
cleave C2 and C4 and activate the
classic pathway.
• Note that this process bypasses the
antibody-requiring step and so is
protective early in infection before
antibody is formed.
Membrane attack complex
• Cleavage of C5 into C5a and C5b.
• C5 (structurally homologous to C3 and
C4, lacks internal thioester bond )
• C5b initiates formation of MAC
(complex of C5b, C6, C7, C8 and
multiple C9 molecules ) binds to C6, and
C7 , recruits C8 and complex penetrates
more deeply into the membrane.
• C9, a pore-forming molecule with
homology to perforin.
• The complex of C5b678 forms a nidus for
C9 binding and polymerization
• Penetrates membrane bilayers to form
pores Disrupt the osmotic barrier, leading to
swelling and lysis of susceptible cells
Biologic Effects of complement
1. Opsonization
• C3b & C1q; enhance phagocytosis
2. Chemotaxis
• C5a and C5,6,7 complex
• attract neutrophils
• C5a – enhance adhesiveness of neutrophils
to the endothelium
3. Anaphylatoxin (C3a, C4a, C5a)
• Cause degranulation of mast cells
• Bind directly to smooth muscles of
bronchioles bronchospasm
4. Cytolysis (MAC)
• Disrupt the membrane & the entry of
water and electrolytes into the cell
5. Enhancement of antibody production
• Binding of C3b to its receptors on the
surface of activated B cells
• enhanced antibody production
C3B is an opsonin
-molecules that bind both to bacteria and
phagocytes
-opsonization increases phagocytosis by
1,000 fold.
Regulation of Complement System
1. C1 inhibitor
 Important regulator of CLASSICAL
PATHWAY.
 A serine protease inhibitor (serpin)
 Irreversibly binds to and inactivates
C1r and C1s, as well as MASP in
lectin pathway
2. Factor H
 Regulate alternative pathway
 Reduce amount of C5 convertase
available
 With both cofactor activity for the
factor I- mediated C3b cleavage, and
decay accelerating activity against
C3bBb (C3 convertase)
3. Properdin
Protects C3b and stabilizes C3 convertase
4. Factor I
 Cleaves cell-bound or fluid phase
C3b and C4b
 Inactivates C3b and C4b
5. Decay accelerating factor
 Glycoprotein on surface of human
cells
 Prevents assembly of performed
convertase  no formation of MAC
Acts on both classical and alternative
6. C4b-binding protein (C4BP)
 Inhibits the action of C4b in
classical pathway
 Splits C4 convertase and is a
cofactor for factor I
7. Complement Receptor 1 (CR-1)
 Co-factor for factor I, together with
CD46
8. Protein (CD59) and Vitronectin (S
protein)
 Inhibits formation of MAC by
binding C5b678
Present on “self” cells to prevent
complement from damaging them.
Clinical Aspects of Complement
1. Deficiency of C5-C8 and Mannan-
binding lectin
 Predispose to severe Neisseria
bacteremia
2. Deficiency of C3
 Severe, recurrent pyogenic sinus and
resp. tract infections
3. Deficiency of C1 esterase inhibitor
 Angioedema  inc. capillary
permeability and edema
4. Deficiency of DAF
 Increased complement-mediated
hemolysis Paroxysmal nocturnal
hemoglobinuria
5. Transfusion mismatches
 Activation of complement 
generate large amounts of
anaphylatoxins and MAC red cell
hemolysis.
6. Autoimmune diseases
 Immune complexes bind
complement  low complement
levels + activate
inflammationtissue damage.
7. Severe liver disease
 Deficient complement proteins
predispose to infection with
pyogenic bacteria
8. Factor I deficiency
 Low levels of C3 in plasma due to
unregulated activation of alternative
pathwayrecurrent bacterial
infections in children.
 Mutations in factor I gene
implicated in development of
HEMOLYTIC UREMIC
SYNDROME.