Antigen Processing and

Presentation
 Antigen Recognition

• T-cells cannot recognise antigen in solution

• Must have the antigen processed and presented
to it
• Specialised molecules present antigens to T cells
• MHC molecules
T cell’s response
Antigen Presentation by MHC
molecules

• Two types of MHC molecules

• MHC class I
• MHC class II
• Two types of T cells
• CD4 T helper cells
• CD8 T cytotoxic cells
• Two types of antigen
• Endogenous (from within the cell)
• Exogenous (from outside the cell)
 Antigen Presentation

• Cells that express MHC class I, present

endogenous ag
• Are killed by CD8+ cells
• Are called Target cells
• Cells that express MHC class II, present
exogenous ag
• Activate the CD4 T cell response
• Are called Antigen presenting cells
 Target & APCs

• Target cells
• All nucleated cells express MHC class I
• Can be a target cell if virally infected/neoplastic
• Antigen presenting cells
• Limited number of cells express MHC class II
• Must sample extracellular environment
• Monocytes, Macrophages, B cells
 Stages of Exogenous
Pathway
• UPTAKE
• Access of native antigens and pathogens to intracellular
pathways of degradation
• DEGRADATION
• Limited proteolysis of antigens to peptides
• ANTIGEN-MHC COMPLEX FORMATION
• Loading of peptides onto MHC molecules
• ANTIGEN PRESENTATION
• Transport and expression of peptide-MHC complexes on the
surface of cells for recognition by T cells
 Endocytic (exogenous)Pathway

• APCs can internalize antigen by phagocytosis &/or

endocytosis.
• Macrophages do both
• B cells use receptor-mediated endocytosis.
• After antigen is internalized, it is degraded into
peptides.
 Exogenous Pathway
• The pathway begins by phagocytosis by the cell of a foreign
agent, an organism, bacteria, etc
• The antigen is now in a phagosome. A lysosome will fuse with
the phagosome to become a phagolysosome.
• The antigen will be degraded into smaller peptides.
• With the help of sorting signals from the invariant chain (that’s
attached to the MHC class 2), the MHC class two will migrate
to the phagolysosome, where it will bind to components that
are 13 – 18 amino acids in size. Once bound, the MHC class 2
will migrate to the membrane to display the antigen.
• A helper T cell will recognize the complex and trigger the
appropriate response, such as secreting cytokines and
chemokines to control whatever kind of infection is taking
place.
 Uptake of exogenous antigens
Membrane Ig
receptor mediated
uptake
Phagocytosis
Y
Complement receptor
mediated phagocytosis

Pinocytosis

Y Fc receptor mediated phagocytosis

Uptake mechanisms direct antigen into intracellular vesicles

for exogenous antigen processing
 Endocytic Pathway

• Internalized antigen takes 1-3 hours to traverse

the endocytic pathway & appear on cell
membrane in the form of peptide-class II MHC
complexes.
• Internalized antigen moves from early to late
endosomes & finally to lysosomes where they are
hydrolyzed into oligopeptides of about 13-18
residues that bind to class II MHCs.
 Exogenous pathway
Cell surface

Protein antigens
Uptake In endosome

Endosomes

Increase
in acidity

To lysosomes

Cathepsin B, D and L proteases are activated by the decrease in pH

Proteases produce peptides from antigens
Drugs that raise the pH of endosomes inhibit antigen processing
MHC class II maturation and invariant chain
In the endoplasmic reticulum

Need to prevent newly Invariant chain stabilises MHC class II by

synthesised, unfolded self non- covalently binding to the immature
proteins from binding to MHC class II molecule and forming a
immature MHC nonomeric complex
 Invariant chain CLIP peptide

 and b chains of MHC class

II molecules

CLIP

A peptide of the invariant chain blocks the MHC molecule binding site.
This peptide is called the CLass II associated Invariant chain Peptide (CLIP)
 Class II associated invariant chain peptide (CLIP)

Cell surface

Endosomes
Uptake

(binv)3 complexes Cathepsin L degrades Invariant MHC Class II

directed towards chain containing vesicles
endosomes by CLIP blocks groove in MHC fuse with antigen
invariant chain molecule containing vesicles
 Removal of CLIP

How can the peptide stably bind to a binding site?

Competition between large number of peptides
 HLA-DM catalyses the removal of CLIP
HLA-DM
Replaces CLIP with a peptide
antigen using a catalytic
mechanism (i.e. efficient at
sub-stoichiometric levels)
Discovered using mutant cell
lines that failed to present
antigen
HLA-DO also play a role in
regulating DM
HLA-DR HLA-DM

Sequence in cytoplasmic tail

retains HLA-DM in endosomes
MIIC compartment
 Surface expression of MHC class II-
peptide complexes
Exported to the cell surface (t1/2 = 50hr)

Sent to lysosomes for degradation

MIIC compartment sorts peptide-MHC complexes for surface expression or

lysosomal degradation
Endogenous Pathway
 Endogenous Pathway
• UPTAKE
• Antigens/pathogens already present in cell
• DEGRADATION
• Antigens synthesised in the cytoplasm undergo limited
proteolytic degradation in the cytoplasm
• ANTIGEN-MHC COMPLEX FORMATION
• Loading of peptide antigens onto MHC class I molecules is
different to the loading of MHC class II molecules
• PRESENTATION
• Transport and expression of antigen-MHC complexes on the
surface of cells for recognition by T cells
 Endogenous Pathway

• We start with an antigen that’s already in the cell. It will be

broken down into smaller peptides by a protease.
• The peptides will be transported into the endoplasmic reticulum
where MHC class 1 is located.
• The 8 – 10 amino acid residues will bind with MHC class 1 and
once that happens, the MHC class 1 and antigen will migrate to
the cell surface, where it will present the antigen.
• Cytotoxic T cells will recognize this complex and initiate the
appropriate immune response to kill this cell.
Peptide antigens produced in the cytoplasm are physically
separated from newly formed MHC class I

ENDOPLASMIC RETICULUM

Newly synthesised
MHC class I molecules

Peptides need
access to the ER in
CYTOSOL order to be loaded onto MHC
class I molecules
 Transporters associated with
antigen processing (TAP1 & 2)
Hydrophobic
transmembrane
Lumen of ER domain
Peptide

ER membrane

Cytosol Peptide
Peptide

Peptide antigens ATP-binding cassette

from proteasome (ABC) domain

Transporter has preference for >8 amino acid peptides

with hydrophobic C termini.
 Maturation and loading of MHC class I

Peptide
Peptide

Peptide

Endoplasmic reticulum

Calnexin binds B2-M Tapasin, calreticulin, TAP 1 Cytoplasmic peptides are

to nascent binds and & 2 form a complex with the loaded onto the MHC
class I chain stabilises floppy MHC molecule and the
until b2-M binds floppy structure becomes
MHC compact
 Fate of MHC class I

Exported to the cell surface

Sent to lysosomes for degradation

Evasion of immunity by interference with endogenous antigen
processing

Peptide

Peptide

Endoplasmic reticulum
Sent to lysosomes
for degradation

HSV protein blocks transport

of viral peptides into ER
Evasion of immunity by interference with
endogenous antigen processing

Normally exported to the cell surface

Adenoviral
protein
retains MHC
class I in the ER

Sent to lysosomes for degradation