2015-10-21

adaptive immunity part 1

lecture 4 adaptive immunity

Adaptive immune system: Adaptive immune system:

introduction introduction
adaptive immune system: primary immune organs
primary lymphatic organs: site of production of B, T, (bone marrow & thymus gland):
NK and other immunocytes
bone marrow: B cells, pre-T cells, NK
antigen-independent development of
thymus gland: T-cells lymphocytes
secondary lymphatic organs
site of most immune responses
lymph nodes, spleen: surrounded by connective tissue capsule
lymphatic nodules: not surrounded by capsule

Adaptive immune system: Adaptive immune system:

introduction introduction

primary immune organs (bone marrow & thymus gland):

bone marrow: major site of hematopoiesis & lymphopoiesis
various blood lineages develop
lymphocytes derived from a common self-renewing hematopoietic
stem cell
HSC: rise to all blood cell lineages
pluripotent stem cell:
common lymphoid progenitor (CLP)
common myeloid progenitor (CMP)
CLP:
progenitor T (pro-T) cell
progenitor B (pro-B) cell
B lymphocytosis: primarly in the bone marrow
T lymphocyte development: thymus at the pro-T stage

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Adaptive immune system: Adaptive immune system:

introduction introduction

secondary lymhoid organs: secondary lymhoid organs:

sites at which the host starts adaptive immune spleen
responses to foreign invaders lymph nodes
MALT
lymphoid follicules (widely distributed)

Adaptive immune system: Adaptive immune system:

introduction introduction

lymph nodes: B cells

encapsulated organ that receives antigens antigen-deriven differentiation
in primary & secondary follicles within the cortex of lymph
from subcutaneous and submucosal tissues nodes
via its afferent lymphatics memory B cells
in the germinal centers of the cortex
T cells
primarily in the diffuse cortex (or paracortex) - associate with
dendritic cells

Adaptive immune system: Adaptive immune system:

introduction introduction
T cells
cells enter the lymph nodes located primarily in
the diffuse cortex
by crossing the high endothelial layer of postcapillary venules (or paracortex),
B cells
located in the diffuse cortex concentrated in where they
primary & associate with
plasma cells secondary follicles dendritic cells
(where they
secreted antibodies exit the lymph nodes via the efferent undergo antigen-
lymphatics and enter the blood stream driven
differentiation)

memory B cells
develop within the
germinal centers
of the cortex

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Adaptive immune system: Adaptive immune system:

introduction introduction
T cells enter the
lymph nodes in spleen:
large numbers by
crossing the high receives antigens through the blood circulation
endothelial layer of
postcapillary contains areas functionally equivalent to those of the
venules located in
the diffuse cortex lymph nodes

terminally
differentiated B secreted antibodies
cells (plasma exit the lymph
cells) nodes via the
efferent lymphatics
& enter the blood
stream

Adaptive immune system: Adaptive immune system:

introduction introduction

Peyer’s patches & submucosa of the small

intestine:
other important peripheral lymphoid tissues
sites in which mucosal antibody responses induced

Adaptive immune system: Adaptive immune system:

introduction introduction

pool of long-lived small lymphocytes

recirculating lymphocytes
continually travel between the various lymphoid
tissues: blood and lymph

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Adaptive immune system: Adaptive immune system:

introduction introduction

recirculating lymphocytes
enter the lymph nodes from the blood
by crossing the high endothelial venules in the diffuse cortex
exit the lymph nodes via the efferent lymphatics
migrate via common thoracic duct to the blood stream
can exit the blood circulation by crossing the endothelium at
many locations

Adaptive immune system: Adaptive immune system:

introduction introduction

body uses both the innate & adaptive

immune systems to combat infections,
however…
… innate immune system eliminates the
majority of infections before any symptoms
begin

Adaptive immune system: Adaptive immune system:

introduction introduction

main characteristics:
specificity:
1 antibody recognizes only 1 antigen
memory:
immune system remembers the infectious agent (antigen)
can prevent it from causing disease in the future
delayed:
slow at first encounter (>96 hours)

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Adaptive immune system: Adaptive immune system:

introduction main characteristics

adaptive immunity distinguishing features: adaptive immunity distinguishing features:

acquired response to antigen, initiated by the primary response to an antigen:
recognition of antigents on foreign invaders several days
host charged by its exposure to antigen, requires:
antigen recognition
becomes „immunized” against a particular antigen
activation & proliferation of T & B lymphocytes
differentiation into populations of effector lymphocytes
antigen-specific memory T & B cells generated
(mediate secondary responses at a later time)
carefully controlled:
long-term maintenance of memory
return of lymphocytes to a nonactivated state

Adaptive immune system: Adaptive immune system:

main characteristics main characteristics

adaptive immunity:
innate adaptive
immunity immunity primary response - lag of approximately 5–7 days
before antibody levels start to rise
response time hours days
time required for:
activation of naive B cells by antigen & Th cells
specificity repeated & fixed highly diversed,
subsequent proliferation
improved during the
differentiation of the activated B cells into plasma cells
course of immune
response in the primary response antibody levels peak at
response to identical to primary much more rapid about day 14
repeated infection infection then primary then drop off (as the plasma cells begin to die)
response

Adaptive immune system: Adaptive immune system:

main characteristics main characteristics
adaptive immunity secondary response
lag much shorter (only 1–2 days)
antibodies:
levels much higher (100x to 1000x higher)
sustained for much longer
reflects the activity of the clonally expanded population of
memory B cells
memory cells respond to the antigen more rapidly
than naive B cells
more memory cells
than there were naive B cells for the primary response
more plasma cells
generated in the secondary response

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Adaptive immune system: Adaptive immune system:

main characteristics main characteristics

primary adaptive response secondary adaptive

response
lag phase primary immune response to an lag period shorter for the secondary
antigen appears only after an initial response
lag phase
amount of smaller greater in the secondary response
antibody

class of primary antibody responses mostly secondary antibody responses consist

immunoglobulin immunoglobulin M (IgM) primarly of non-IgM classes, especially
IgG

change in the class of antibody produced = isotype switching = results in new functions associated
with the same antibody specificity

duration shorter longer

longer duration of the secondary resonse = large number of memory B cells actived and the
longer half-life of IgG compared to IgM

Adaptive immune system: Adaptive immune system:

main characteristics BCR and TCR

adaptive immunity: T and B lymphocytes:

immunity mediated by lymphocytes or the antibodies they primary mediators of adaptive immunity
produce can be transferred from an immune host to a recognize antigenic determinants by their cell surface
naive recipient antigen receptors
transfer of antibodies called: passive immunization receptors with specificity for autoantigens expressed
transfer of passive immunity from the mother to the neonate during the development of the adaptive immune
transfer of maternal serum IgG from the mother to the offspring system
in utero
most lymphocytes with autoreactive receptors deleted
after birth (ingestion of immunoglobulin-rich colostrum by the some autoreactive lymphocytes survive (their activation carefully
neonate) controlled)
adaptive immunization: transfer of immune cells

Adaptive immune system: Adaptive immune system:

BCR and TCR BCR and TCR

B cell receptors predominantly recognize soluble BCR

native antigens membrane-bound antibodies & antibodies that B
T cell receptors recognize foreign antigens only on cells secrete
the surface of APC able to recognize:
many of the effector cells and molecules in adaptive proteins
lipids
immunity - the same as in innate immunity
carbohydrates
nucleic acids
simple small chemical groups
parts of macromolecules

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Adaptive immune system: Adaptive immune system:

BCR and TCR BCR and TCR

BCR BCR
immunoglobulin associated with 'accessory' heterodimers interact with the transmembrane parts of the
molecules immunoglobulin receptor
on the B cell surface cellular activation (like the separate molecular components of the
'B cell antigen receptor complex' (BCR) TCR/CD3 complex)

'accessory' molecules consist of disulfide-bonded intracellular domains of CD79a/b have immunoreceptor

tyrosine-based activation motifs (ITAMs)
heterodimers of:
Igα (CD79a) BCR interaction with specific antigen:
ITAM phosphorylation
Igβ (CD79b)
downstream cascade of the activation-related changes in gene
expression

Adaptive immune system: Adaptive immune system:

BCR and TCR BCR and TCR

BCR
two identical heavy (H) chains & two identical light (L) chains
secondary components (Igα and Igβ)
closely associated with the primary receptor
thought to couple it to intracellular signaling pathways
circulating antibodies structurally identical
to the primary B cell antigen receptors
lack the transmembrane & intracytoplasmic sections

Adaptive immune system: Adaptive immune system:

BCR and TCR BCR and TCR

T cells can „see” in general: antigen-presenting cells (APCs):

only peptides specialized system for
capturing antigen
only when these peptides displayed on antigen presenting
bringing to the organs where T cell responses initiated
cells (APCs)
bound to membrane proteins dendritic cells especially important for activating naive T cells
encoded in the major histocompatibility complex (MHC) gene present in all tissues
locus capture antigens
TCRs exist only as membrane receptors of T cells migrate to the same regions of lymphoid organs where
recirculating T cells localize
maximizing the chance of T cells of a particular specificity finding the
antigen

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Adaptive immune system: Adaptive immune system:

BCR and TCR TCR

some types of T lymphocytes require interaction with: T cell antigen receptors designed to see antigens
dendritic cells displayed by host cell surface molecules
macrophages
B lymphocytes T cells (in general) interact only with other host cells
but:
other types of T lymphocytes must be able to interact not directly with microbes
with: not with antigens on microbes
infected host cell not with antigens that are free in the circulation or
extracellular fluids

Adaptive immune system: Adaptive immune system:

TCR TCR

B lymphocytes receptor (BCR) displaying host cell-associated antigens

antigen receptors & secreted products antibodies, for recognition by CD4+ and CD8+ T cells performed
can recognize antigens on: by
microbial surfaces
soluble antigens major histocompatibility complex (MHC)
cell-associated antigens molecules

expressed on the surfaces of host cells

Adaptive immune system: Adaptive immune system:

BCR and TCR BCR and TCR

T cells respond to antigens in different compartments

T cells defense against viruses

in the circulation
mediated by antibodies
production of antibodies requires CD4+ helper T cells
in the tissue cells
usually inaccessible to the antibody
eradication requires CD8+ cytotoxic T lymphocytes (CTLs)

APCs handle antigens in extracellular & intracellular locations

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Adaptive immune system: Adaptive immune system:

BCR and TCR BCR and TCR

B cells can recognize T lymphocytes generally recognize only short

peptides linear peptides
proteins haptens
can bind to self proteins
nucleic acids
hapten-conjugated peptides might be recognized by T cells
carbohydrates some T cells specific for small chemical haptens such as:
lipids dinitrophenol
small chemicals urushiol of poison ivy
β−lactams of penicillin antibiotics
peptide specificity of T cells is true for CD4+ and
CD8+ cells
small populations of T cells capable of recognizing
nonprotein antigens

Adaptive immune system: Adaptive immune system:

TCR TCR
Features of Antigens Recognized by T Cells Explanation
T cells recognize only peptides:
antigen receptors of CD4+ & CD8+ T cells specific for
Most T cells recognize peptides and no other Only peptides bind to MHC molecules.
molecules.
antigens displayed by MHC molecules
MHC can bind peptides but no other chemical structures
T cells recognize linear peptides & Linear peptides bind to clefts of MHC molecules,
not conformational determinants of protein & protein conformation is lost during the
MHC restriction:
antigens. generation of these peptides. single T cell recognize a specific peptide displayed by
T cells recognize cell-associated &
not soluble antigens.
T cell receptors recognize only MHC-like shapes,
and MHC molecules are membrane proteins that
only 1 of the large number of different MHC molecules
display stably bound peptides on cell surfaces.
CD4+ and CD8+ T cells preferentially Pathways of assembly of MHC molecules ensure
recognize antigens sampled from the that class II molecules display peptides derived
extracellular and cytosolic pools, respectively. from extracellular proteins and taken up into
vesicles in APCs and that class I molecules
present peptides from cytosolic proteins;
CD4 and CD8 bind to nonpolymorphic regions of
class II and class I MHC molecules, respectively.

Adaptive immune system: Adaptive immune system:

TCR BCR and TCR

antigen–specific T cell receptor (TCR):

αβ or γδ heterodimer polypeptides
each polypeptide of TCR contains variable &
constant domains
genetically & molecularly distinct from immunoglobulins
choice of αβ or γδ heterodimer made early in T cell
development

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Adaptive immune system: Adaptive immune system:

TCR BCR and TCR

TCR bound to the membrane of T cell

short cytoplasmatic tails of αβ or γδ chains

unable to initiate activation signal to nucleus due to lack of:
signaling sequences
immunoreceptor tyrosine activation motifs (ITAMs)

activation signal to nucleus provided by CD3 complex

molecules (CD3γ, CD3δ, CD3ε and CD247 – ζ chain)
noncovalently associate with TCR

Adaptive immune system: Adaptive immune system:

TCR TCR

TCR: TCR variable & constant regions:

TCR cannot bind soluble epitopes each polypeptide chain of TCR contains:
variable region domain (Vα or Vβ; Vγ or Vδ)
(unlike antibodies)
constant region domain (Cα or Cβ; Cγ or Cδ)
TCR can bind only to fragment of larger together variable regions of α and β (or γ and δ) chains form
molecules within the groove of MHC class I or hypervariable or complimentary-determining regions
MHC class II molecules (CDRs)
peptide-MHC (pMHC) complexes
interaction of the TCR with pMHC: each T cell
stabilized by associated interaction of CD4 or CD8 with expresses a unique TCR (like immunoglobulins)
constant domains of MHC class II or MHC class I must „see” pMHC
molecules do not recognize soluble peptides (unlike
immunoglobulins)

Adaptive immune system:

TCR

part 2

APC
antigen presenting cell

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Adaptive immune system: Adaptive immune system:

APC APC

APC: dendritic cells, macrophages & B lymphocytes

different cells which can activate naive & previously capable of activating CD4+ T lymphocytes because
differentiated effector T cells they express:
class II MHC molecules
dendritic cells: most effective APCs for activating
other molecules involved in stimulating T cells
naive T cells
macrophages & B lymphocytes also function as 3 above cell types: professional APCs
APCs (term sometimes used to dendritic cells only)
mostly for previously activated CD4+ helper T cells only cell dedicated to capture & present antigens
(rather than for naive T cells)
only APC capable of initiating primary T cell responses

Adaptive immune system: Adaptive immune system:

APC APC

APC:
display peptide-MHC complexes for T cells
provide additional stimuli (second signals) required
for the full responses of the T cells
second signals more important for activation of naive T
cells than for previously activated effector & memory cells
membrane-bound molecules of APCs that serve to
activate T cells: costimulators

critical roles in T cell differentiation into effector

cells: secreted cytokines

Adaptive immune system: Adaptive immune system:

APC APC

APC: APC:
antigen-presenting function enhanced by dendritic cells activated by microbes express
exposure to microbial products chemokine receptors
dendritic cells & macrophages express (stimulate migration to sites with T cells)
Toll-like receptors optimal T cell responses to protein antigens
other microbial sensors
requires antigens with adjuvants
respond to microbes
products of microbes (killed mycobacteria)
increasing the expression of MHC molecules & costimulators
products that mimic microbes & enhance the expression of
improving the efficiency of antigen presentation
costimulators & cytokines
activating the APCs to produce cytokines

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Adaptive immune system: Adaptive immune system:

APC APC

APC: APC:
APCs presenting antigens to T cells combination of CD40 signals & cytokines activates
receive signals from T cells the APCs
(enhance antigen-presenting function) increased ability to process & present antigens
increased expression of costimulators
secretion of cytokines that activate the T cells
CD4+ T cells activated by antigen recognition &
costimulation express surface molecule
CD40 ligand (CD154) - binds to CD40 bidirectional interaction
on dendritic cells & macrophages between APCs displaying the antigen and
T lymphocytes that recognize the antigen
T cells secrete cytokines such as interferon-γ (IFN-γ) that functions as a
bind to their receptors on these APCs positive feedback loop

Adaptive immune system: Adaptive immune system:

properties and functions of APC APC
cell type class II MHC expression of principal function
costimulators role of dendritic cells in antigen capture & display:
dendritic cells constitutive; increases with constitutive; increases with initiation of T cell responses
primary responses of naive CD4+ T cells initiated in the
maturation; increased by maturation; inducible by to protein antigens peripheral lymphoid organs
IFN-γ IFN-γ, CD40-CD40L (priming)
interactions protein antigens transported to peripheral lymphoid organs
macrophages low or negative; inducible inducible by LPS, IFN-γ, effector phase of cell- after being collected from
by IFN-γ CD40-CD40L interactions mediated immune
responses (T cell-enhanced
skin
killing of phagocytosed epithelia of the GI & respiratory systems
microbes)
B lymphocytes constitutive; increased by induced by T cells (CD40- antigen presentation to
antigens
IL-4 CD40L interactions), CD4+ helper T cells in produced in any tissue colonized or infected by a microbe
antigen receptor cross- humoral immune responses
linking (cognate T cell-B cell concentrated in lymph nodes
interactions) filters that sample the lymph before it reaches the blood
vascular endothelial cells inducible by IFN-γ; constitutive (inducible in may promote activation of in blood may be similarly sampled by the spleen
constitutive in humans mice) antigen-specific T cells at
site of antigen exposure
various epithelial and inducible by IFN-γ probably none no known physiologic
mesenchymal cells function

Adaptive immune system: Adaptive immune system:

APC APC

morphology & populations of dendritic cells:

DCs arise from bone marrow precursors
most related in lineage to mononuclear phagocytes
several subsets of DCs
distinguished by the expression of various cell surface markers
play different roles in immune responses
two main types:
conventional DCs
plasmacytoid DCs

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Adaptive immune system: Adaptive immune system:

APC APC
conventional DCs tissue-derived conventional DCs:
identified by morphology & ability to stimulate T cells Langerhans cell type
most numerous DC
DCs in epithelia
from bone marrow progenitors
functions: skin-draining lymph nodes
resident tissue population of DCs Langerhans cells of the epidermis
on activation (with microbes or cytokines) mature & migrate occupy as much as 25% of the surface area of the epidermis
into draining lymph nodes
constitute less than 1% of the cell population
initiate T cell responses
interstitial/dermal type
most other tissues

Adaptive immune system: Adaptive immune system:

APC APC

conventional DCs plasmacytoid DCs

surface markers: morphologically resemble plasma cells
CD11c high only after activation acquire morphology & functional
CD11b high properties of DCs
growth factors for in vitro derivation: develop in the bone marrow
GM-CSF, Flt3-ligand found:
expression of Toll-like receptors (TLRs): blood
TLRs 4, 5, 8 high lymphoid organs (T cell zones of the spleen and lymph nodes)
major cytokines produced:
TNF, IL-6 major function of plasmacytoid DCs
postulated major function: secretion of large amounts of type I interferons in response to
induction of T cell responses against most antigens viral infections
play a role in presenting antigens to T lymphocytes

Adaptive immune system: Adaptive immune system:

APC antigen capture & transport by dendritic cells

plasmacytoid DCs
surface markers:
CD11c low
CD11b negative
B220 high
growth factors for in vitro derivation:
Flt3 ligand
expression of Toll-like receptors (TLRs):
TLRs 7, 9 high
major cytokines produced:
type I interferons
postulated major function:
innate immunity and induction of T cell responses against viruses

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Adaptive immune system: Adaptive immune system:

antigen capture & transport by dendritic cells antigen capture and transport by dendritic cells

DCs resident in epithelia & tissues: DCs innate immune response:

capture protein antigens activated by:
transport the antigens to draining lymph nodes microbial products recognized by TLRs in the DCs & other cells
cytokines (i.e. TNF)

resting (immature) DCs express membrane

activated DCs („mature” DCs):
receptors (i.e. C-type lectins) that bind microbes lose their adhesiveness for epithelia or tissues
process ingested proteins into peptides capable of binding to migrate into lymph nodes
MHC molecules
express a CCR7 chemokine receptor (specific for
receptor-mediated endocytosis & phagocytosis
chemokines CCL19 & CCL21) produced in the T cell zones of
micropinocytosis & macropinocytosis
lymph nodes
chemokines attract the DCs with microbial antigens into the T
cell zones of the lymph nodes

Adaptive immune system: Adaptive immune system:

antigen capture and transport by dendritic cells antigen capture & transport by dendritic cells

naive T cells antigens:

also express CCR7 may be transported to lymphoid organs in soluble form of
maximize the chance of T cells with receptors for the antigen find that lymph- & blood-borne antigens
antigen may be captured by resident DCs in the lymph nodes & spleen
migrate to the same regions of lymph nodes where antigen-bearing lymph enters a lymph node through an afferent lymphatic vessel
DCs concentrated drains into the subcapsular sinus

activated „mature” DCs:

B cells in the node may also recognize & internalize soluble
express high levels of MHC molecules with peptides & costimulators
antigens
become resident in lymph nodes (turned into APCs: ability to activate
T lymphocytes)
DCs, macrophages, and B cells present antigens to:
naive T cells recirculating through lymph nodes eventually
naive T cells
encounter APCs - T cells specific for peptide-MHC complexes
effector T cells (generated by previous antigen stimulation)
become activated

Adaptive immune system: Adaptive immune system:

antigen capture and transport by dendritic cells antigen capture and transport by dendritic cells

DCs: the most efficient APCs for initiation of primary

T cell responses
located at the sites of entry of microbes & foreign
antigens
(in epithelia or tissues)
express receptors enabling to capture
& respond to microbes
migrate to the T cell zones of lymph nodes
(where naive T lymphocytes circulate)
mature DCs express high levels of peptide-MHC
complexes, costimulators, and cytokines
(needed to activate naive T lymphocytes)

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Adaptive immune system: Adaptive immune system:

antigen capture & transport by dendritic cells APC

DCs: functions of other APCs:

can ingest infected cells macrophages:
present antigens from these cells to CD8+ T lymphocytes present the antigens of phagocytosed microbes to effector
CD8+ lymphocytes recognize antigens produced in T cells
respond by activating the macrophages to kill the microbes
any cell type infected by a virus (in cell-mediated immune response)
not necessarily DCs B lymphocytes:
specialized DCs: internalize protein antigens & derived present peptides to
helper T cells
ability to ingest virus-infected cells or cellular fragments
in humoral immune responses
present antigens to CD8+ T lymphocytes
process is called cross-presentation (cross-priming)

Adaptive immune system:

APC

functions of other APCs:

all nucleated cells can present peptides derived from
cytosolic protein antigens to CD8+ T lymphocytes
part 3
phagocytosed microbes may be recognized by CD8+ CTLs
if these microbes or their antigens escape
from phagocytic vesicles into the cytosol

vascular endothelial cells in humans express class II MHC MHC

molecules
present antigens to blood T cells
contribute to the recruitment & activation of effector T cells

Adaptive immune system: Adaptive immune system:

MHC MHC

major histocompatibility complex (MHC) = human major histocompatibility complex (MHC) = human leukocyte antigen
leukocyte antigen (HLA): (HLA)
segment of chromosome 6 containing several genes critical to
immune function MHC class I:
codominantly expressed 45 kDa molecules
include genes encoding various enzymes & structural
together with β2 microglobulin (12 kDa) found on surfaces of all
molecules needed for activation & function of B & T nucleated cells
lymphocytes include 3 genes called: HLA-A, HLA-B, HLA-C
3 groups (or classes) known as MHC class I, II & III molecules encode 3 class I MHC molecules with the same names
MHC class III molecules include complement components C4, Bf
3 genetic loci highly polymorphic: >100 alleles at each locus
and C2
up to 6 different class I molecules (if heterozygous at all three loci) can
be simultaneously displayed on each cell

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Adaptive immune system: Adaptive immune system:

MHC MHC

class I MHC: class II MHC molecules normally expressed on surfaces:

fold to form a cleft (groove) between α1 and α2 domains: dentritic cells
non-covalently binds 8 to 9-amino-acid peptide macrophages
B-lymphocytes
additional („non-classical” or class Ib) class I some activated T-lymphocytes
molecules: some specialized epithelial cells in thymus & intestine
HLA-E, -F, -G, -H loci 3 class II HLA gene loci called HLA-DP, HLA-DQ, and HLA-DR
limited variability & tissue distribution each class II MHC molecule
present carbohydrate & peptide fragments composed of a heterodimer of α and β polypeptides
DP, DQ, and DR loci each contain separate genes designated A or B,
encoding α and β chains

Adaptive immune system: Adaptive immune system:

MHC MHC

class II HLA molecules: class II HLA molecules:

codominantly expressed as noncovalent heterodimers: after synthesis MHC class II molecules
α chain and β chain form a binding groove (α1 and β1 domains) α and β chain combines only with the others encoded by
can bind peptides of 18 do 20-aminoacids lenght the same region
(eg. DPα associates only with DPβ but never with DQβ or
encoded within HLA-DP, HLA-DQ and HLA-DR regions are
DRβ etc.)
both α and β loci (DPα, DPβ, DQα, DQβ, etc.)
α chains may combine either with β chain encoded on the same
chromosome (cis) or on the member of the chromosome pair
(trans)
termed cis-trans complementation

Adaptive immune system: Adaptive immune system:

MHC MHC

MHC genes codominantly expressed in

each individual
for a given MHC gene, each individual expresses
the alleles inherited from each of the two
parents
maximizes nr of MHC molecules available to bind
peptides for T cells presentation

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Adaptive immune system: Adaptive immune system:

MHC MHC

MHC locus contains two types of polymorphic class I MHC molecules display peptides to
MHC genes: class I and class II MHC genes CD8+ T cells
encode 2 groups of structurally distinct but homologous class II MHC molecules display peptides to
proteins CD4+ T cells
other nonpolymorphic genes whose products involved in
antigen presentation each of these T cell types serves different functions in
protection against microbes

Adaptive immune system: Adaptive immune system:

MHC MHC

MHC class I: MHC class II:

function of class I-restricted CD8+ CTLs: kill cells class II-restricted CD4+ helper T lymphocytes require
infected with intracellular microbes, such as viruses recognizing antigen presented by a more limited number of
cell types
tumors that express tumor antigens
naive CD4+ T cells
expression of class I MHC molecules on antigens captured & presented by dendritic cells in lymphoid
nucleated cells serves provides organs

a display system for viral & tumor antigens differentiated CD4+ helper T lymphocytes
activate (or help) macrophages to eliminate extracellular
phagocytosed microbes
activate B lymphocytes to make antibodies against extracellular
microbes
class II molecules
expressed mainly on these cell types
display of peptides derived from extracellular microbes & proteins

Adaptive immune system: Adaptive immune system:

MHC MHC

expression of MHC molecules increased by expression of MHC molecules increased by cytokines

cytokines MHC class II
produced during both innate & adaptive immune responses IFN-γ:
interferons: produced during the early innate immune principal cytokine involved in
response to viruses stimulating expression of MHC class II in APCs
MHC class I dendritic cells
macrophages
interferons IFN-α, IFN-β, and IFN-γ increase the level of
vascular endothelial cells & other nonimmune cell types
expression of MHC class I molecules
IFNs may be produced by
NK cells
during innate immune reactions
antigen-activated T cells
during adaptive immune reactions

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Adaptive immune system: Adaptive immune system:

MHC MHC

MHC class II expression increases in response to: expression of MHC molecules

signals from Toll-like receptors MHC molecule synthesis & expression on the cell surface
responding to microbial components dependant on rate of transcription
B lymphocytes protein: class II transcription activator (CIITA)
constitutively express class II molecules
synthesized in response to IFN-γ
can increase expression in response to
antigen recognition & cytokines produced by helper T cells mutations in transcription factors cause of
human immunodeficiency diseases
with defective expression of MHC: bare lymphocyte syndrome

Adaptive immune system: Adaptive immune system:

MHC MHC

general properties of MHC molecules:

MHC molecule:
an extracellular peptide-binding cleft (or groove)
immunoglobulin (Ig)-like domains
transmembrane domain
cytoplasmic domains

Adaptive immune system: Adaptive immune system:

MHC MHC

general properties of MHC molecules: general properties of MHC molecules:

polymorphic amino acid residues CD4 binds selectively to class II MHC molecules
of MHC molecules located in and adjacent to CD4+ helper T cells recognize class II MHC
peptide-binding cleft molecules displaying peptides
nonpolymorphic Ig-like domains of MHC molecules
contain binding sites for the T cell molecules CD4 CD8 binds to class I MHC molecules
and CD8 CD8+ T cells recognize class I MHC molecules with
CD4 and CD8 expressed on distinct subpopulations of bound peptides
mature T lymphocytes

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Adaptive immune system: Adaptive immune system:

MHC MHC

general properties of MHC molecules: class I MHC molecules consist of

2 noncovalently linked polypeptide chains
CD4+ T cells are class II MHC restricted α chain (or heavy chain) MHC-encoded 44- to 47-kD
CD8+ T cells are class I MHC restricted β2-microglobulin (non-MHC-encoded 12-kD subunit)
peptide-binding cleft of MHC class I molecules
bind peptides of 8 to 11 amino acids
ends of the class I peptide-binding cleft are closed
(larger peptides have to be "processed" to fragments small enough)
α3 segment folds into an Ig domain
amino acid sequence conserved among all MHC class I molecules
α3 segment contains the binding site for CD8
β2-microglobulin (the light chain of class I molecules)
encoded by a gene outside the MHC
interacts noncovalently with the α3 domain of the α chain
structurally homologous to an Ig domain
invariant among all class I molecules

Adaptive immune system: Adaptive immune system:

MHC MHC

class I MHC molecules

fully assembled class I molecule: heterotrimer
consisting of 3 parts:
α chain
β2-microglobulin
bound antigenic peptide
stable expression of MHC class I molecules - presence of all 3
components of the heterotrimer

Adaptive immune system: Adaptive immune system:

MHC MHC

class II MHC molecules class II MHC molecules: amino-terminal α1 and β1

composed of 2 noncovalently associated polypeptide chains segments of the class II MHC form the peptide-
α chain (32- to 34-kD)
β chain (29- to 32-kD)
binding cleft
unlike class I molecules, the genes encoding both chains of class structurally similar to the cleft of MHC class I molecules
4 strands of the floor of the cleft & 1 of the α-helical walls are
II molecules are polymorphic and present in the MHC locus
formed by the α1 segment
other 4 strands of the floor & the second wall are formed by the
β1 segment
ends of the peptide-binding cleft open:
peptides of 30 residues or more can fit

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Adaptive immune system: Adaptive immune system:

MHC MHC

class II MHC molecules

β2 segment of class II MHC contains the binding site for
CD4
similar to the binding site for CD8 in the α3 segment of the class I
heavy chain
fully assembled class II molecule heterotrimer consisting of:
α chain
β chain
bound antigenic peptide
stable expression of class II molecules on cell surfaces
requires the presence of all three components of the
heterotrimer

Adaptive immune system: Adaptive immune system:

MHC characteristics of peptide-MHC interactions

MHC molecules show a broad specificity for peptide

binding
in contrast to the specificity of the antigen receptors of
lymphocytes
class I or class II MHC molecule has a single
peptide-binding cleft that binds one peptide at a
time
each MHC molecule can bind many different
peptides
peptides that bind to MHC molecules share
structural features that promote this interaction

Adaptive immune system: Adaptive immune system:

characteristics of peptide-MHC interactions MHC
MHC molecules acquire their peptide cargo during
their biosynthesis & assembly inside cells
association of antigenic peptides and MHC molecules
- saturable interaction with a very slow off-rate
very small numbers of peptide-MHC complexes
capable of activating specific T lymphocyte
MHC molecules do not discriminate between
foreign peptides (e.g., those derived from microbial
proteins) and peptides derived from self antigens

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Adaptive immune system: Adaptive immune system:

MHC MHC

processing of peptide antigens processing of peptide antigens:

pathways of antigen processing protein antigens
convert protein antigens present in the cytosol (synthesized in the cell) generate
present in the cytosol or internalized from the MHC class I peptides
extracellular environment recognized by CD8+ T cells
into peptides
internalized from the extracellular environment into the
load these peptides onto MHC molecules vesicles of APCs generate peptides displayed by
for display to T lymphocytes MHC class II molecules
recognized by CD4+ T cells

Adaptive immune system: Adaptive immune system:

MHC MHC

class I MHC pathway:

most cytosolic protein antigens synthesized within cells (some
phagocytosed & transported into the cytosol)
products of viruses
products of intracellular microbes (that infect such cells)
some microbes damage phagosome membranes & antigens enter the cytosol
Listeria monocytogenes (listeriolysin)
mutated or overexpressed genes protein antigens in tumor cells
in dendritic cells, some antigens ingested into vesicles enter the
cytosolic class I pathway - process called cross-presentation
other sources of peptides in the cytosol
misfolded proteins in the ER (ER-associated degradation)

Adaptive immune system: Adaptive immune system:

MHC MHC

class I MHC pathway:

proteolytic digestion of cytosolic proteins
proteolysis - major mechanism for the generation of
peptides from cytosolic protein antigens
proteasome: multiprotein enzyme complexes with
proteolytic activity found in the cytoplasm & nuclei of most
cells
two inner β rings (3 of 7 subunits β1, β2, β5): catalytic
two outer α rings: structural
ubiquitin
INF-γ (increased transcription & synthesis of 3 catalytic
subunits: β1i, β2i, β5i)

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Adaptive immune system: Adaptive immune system:

MHC MHC

class I MHC pathway: transport of peptides from the class I MHC pathway for processing and presentation of cytosolic
proteins:
cytosol to ER assembly of peptide-class I MHC complexes in the ER
peptides generated in the cytosol translocated by a peptides translocated into the ER bind to class I MHC molecules
specialized transporter into the ER associated with TAP through tapasin
where newly synthesized available class I MHC molecules chaperone proteins (membrane: calnexin, luminal chaperone: calreticulin)
newly formed empty class I dimers remain linked to the TAP complex
mediated by protein: transporter associated with antigen
empty class I MHC molecules, tapasin, and TAP part of a larger peptide-
processing (TAP) loading complex (also calnexin, calreticulin, & the oxidoreductase Erp57, all
TAP protein located in the ER membrane take part in assembly & loading)
peptides that enter the ER through TAP & peptides produced in the ER (as
mediates ATP transport of peptides from the cytosol into the ER signal peptides) often trimmed to the appropriate size for MHC binding by the
optimally transports peptides from 8 to 16 amino acids and ER-resident aminopeptidase ERAP
containing carboxyl termini class I MHC molecules loaded with peptide, peptide-class I complex is
on the luminal side of the ER membrane, TAP protein associates released from tapasin, able to exit the ER and be transported to the cell
surface
with a protein called tapasin
peptides transported into the ER preferentially bind to MHC class I
affinity for newly synthesized empty class I MHC molecules but not class II MHC molecules
tapasin brings TAP transporter into a complex with the class I
MHC molecules

Adaptive immune system: Adaptive immune system:

MHC MHC

class I MHC pathway for processing & presentation of cytosolic

proteins:
surface expression of peptide-class I MHC complexes
class I MHC molecules with bound peptides structurally stable
& expressed on the cell surface
may be recognized by peptide antigen-specific CD8+ T cells

Adaptive immune system: Adaptive immune system:

MHC MHC

class II MHC pathway for processing and presentation

of vesicular proteins
most class II-associated peptides derived from
protein antigens
captured from extracellular environment
internalized into endosomes by specialized APCs

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Adaptive immune system: Adaptive immune system:

MHC MHC

class II MHC pathway for processing and presentation class II MHC pathway for processing and presentation
of vesicular proteins of vesicular proteins
different APCs bind protein antigens: internalization:
dendritic cells & macrophages: surface receptors protein antigens in intracellular membrane-bound vesicles called
recognizing structures shared by many microbes endosomes
macrophages have receptors for: communicates with lysosomes
Fc portions of antibodies microbes internalized into vesicles called phagosomes, may fuse
complement protein C3b with lysosomes, producing phagolysosomes or secondary
B cells: surface immunoglobulin lysosomes
high affinity for antigens some microbes (Mycobacterium and Leishmania) may survive and
mediate the internalization of proteins present at very low replicate within phagosomes or endosomes
concentrations in the extracellular fluid

Adaptive immune system: Adaptive immune system:

MHC MHC

class II MHC pathway: class II MHC pathway:

α and β chains of class II MHC molecules synthesized &
internalized proteins degraded enzymatically in late
associate with each other in the ER
endosomes & lysosomes
structurally unstable
generated peptides able to bind to the peptide-binding clefts folding & assembly aided by ER-resident chaperones (calnexin)
of class II MHC molecules invariant chain (Ii):
process mediated by proteases promotes folding & assembly of class II molecules
cathepsins (thiol & aspartyl proteases) directs newly formed class II molecules to the late endosomes &
lysosomes
partially degraded or cleaved proteins bind to the open-ended clefts
invariant chain (Ii): trimer composed of 30-kD subunits, 3 of which bind one
of class II MHC molecules and cut enzymatically to their final size newly synthesized class II αβ heterodimer = blocks the peptide-binding cleft
class II MHC molecules class II MHC molecules transported in exocytic vesicles toward the cell
synthesized in the ER surface
vesicles taking class II molecules out of the ER fuse with the
transported to endosomes with an associated protein, the
endocytic vesicles containing antigens
invariant chain (Ii)
peptide-MHC association occurs in the vesicles
occupies the peptide-binding clefts of the newly synthesized class II
molecules

Adaptive immune system: Adaptive immune system:

MHC MHC
class II MHC pathway:
invariant chain (Ii) dissociates from class II MHC molecules due class II MHC pathway:
to action of proteolytic enzymes and the HLA-DM molecule expression of peptide-class II MHC complexes on the cell
antigenic peptides then able to bind to the available peptide-binding surface
clefts of the class II MHC
stabilized by the bound peptides
invariant chain removed before complexes of peptide & MHC class II
form stable peptide-class II complexes delivered to the surface of the
proteolytic enzymes (i.e.cathepsins) act on the Ii, degrading it & leaving only a
APC
24-amino acid remnant called class II-associated invariant chain peptide displayed for recognition by CD4+ T cells
(CLIP)
CD4 coreceptor playing an essential role by binding to
CLIP has to be removed so that the cleft becomes accessible to nonpolymorphic regions of the class II molecule
antigenic peptides produced from extracellular proteins.
molecules involved in antigen presentation, such as DM, stay in
removal is due to a molecule called HLA-DM
the vesicles and not expressed in the plasma membrane
HLA-DM molecules differ from class II MHC molecules:
not polymorphic
not expressed on the cell surface
acts as a peptide exchanger

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Adaptive immune system: Adaptive immune system:

MHC MHC

class II MHC pathway:

cross-presentation: dendritic cells - ability to capture &
ingest virus-infected cells (or tumor cells) & present the
viral or tumor antigens to naive CD8+ T lymphocytes
ingested antigens transported from vesicles to the cytosol, from
where peptides enter the class I pathway
most ingested proteins do not enter the cytosolic class I
pathway
protein traffic from endosomal vesicles to the cytosol unique to
dendritic cells
process called cross-presentation or cross-priming
one cell type (dendritic cell) can present antigens from another
cell (virus-infected or tumor cell) & prime (activate) T cells
specific for these antigens

Adaptive immune system: Adaptive immune system:

MHC MHC

physiologic significance of MHC antigen presentation: physiologic significance of MHC antigen presentation:
presentation of cytosolic vs. vesicular proteins by the class I or extracellular antigens (endosomal vesicles) activate class II-
class II MHC, determines which subsets of T cells will restricted CD4+ T cells
respond to antigens vesicular proteins processed into class II-binding peptides
CD4+ T cells function as helpers to:
endogenously synthesized antigens (viral & tumor) located B cells (to produce antibodies)
in the cytoplasm & recognized by class I-restricted CD8+ macrophages (to enhance their phagocytic activity)
CTLs
CTLs kill the cells producing the intracellular antigens

Adaptive immune system: Adaptive immune system:

MHC MHC

presentation of nonprotein antigens to subsets

of T cells:
small populations of T cells able to recognize
nonprotein antigens without class I or class II MHC
molecules:
NKT
γδT cells

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Adaptive immune system: Adaptive immune system:

MHC MHC

presentation of nonprotein antigens to subsets of T presentation of nonprotein antigens to subsets of T

cells: NKT cells cells: NKT cells
both natural killer (NK) cells & T lymphocytes markers several CD1 proteins expressed in humans and mice
express αβTCR with very limited diversity all the CD1 molecules bind & display lipids by a unique
recognize lipids & glycolipids displayed by the pathway
class I-like "non-classical" MHC molecule called CD1 CD1 pick up cellular lipids & carry these to the cell surface
CD1-lipid complexes - into endosomes or lysosomes
CD1 molecules acquire endocytosed lipid antigens during recycling &
present antigens without apparent processing
NKT cells that recognize the lipid antigens may play a role
in defense against microbes (especially Mycobacterium)

Adaptive immune system:

MHC

presentation of nonprotein antigens to subsets of T cells:

γδT cells:
small population of T cells
express antigen receptor proteins similar
Thank you
but not identical to those of CD4+ and CD8+ T cells
recognize many different types of antigens
proteins
lipids
small phosphorylated molecules
alkyl amines
antigens not displayed by MHC molecules, so
γδT cells are not MHC restricted
not known if a particular cell type or antigen display system
required for presenting antigens to these cells

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