Activation and Function of T and B

Cells
 Interaction of Ag with Ag-specific receptors on B or T
cells initiates a cascade of events that results in the
cells proliferation and further differentiation
 Differentiate into
 Effector cells
 Memory cells
 B cells  Ab production
 T cells 
 produce cytokines that affect different cell types
 Become directly cytotoxic
TCR
T Dev
T Helper Cell Surface Molecules
Activation of CD4+ T Cells
 APCs (such as dendritic cells) present peptide + class II MHC to
CD4+ T cells in the primary response
 B cells are likely to be the major APC in the response of primed
(memory) T cells
 V + V of the TCR on the CD4+ T cell recognize
 Peptide + class II MHC
 Contact b/w TCR and MHC is necessary but generally not
sufficient for T cell activation
 Likely due to low affinity of the interaction b/w TCR and peptide-
MHC
 Also due to small number of peptide –MHC presented on the APC
(as few as 100 per cell)
T cell
Activation
Activation of a T4-Lymphocyte by an APC
Subsets of CD4+ Cells Defined by
Cytokine Production
 Synthesis of Ag-nonspecific cytokines is a key
effector function of activated CD4+ cells
 Cytokines produced by CD4+ cells affect many cells
 T cells, B cells, myeloid cells (macrophages and
eosinophils), bone marrow precursors
 For this reason, the loss of CD4+ T cells in AIDS is so
devastating
 Ag-primed CD4+ human T cells can be divided into
three subsets
 TH0, TH1 and TH2
Differentiation of TH1 and TH2 Cells
 TH0 cells synthesize IL-2, IFN- and IL-4
 TH1 and TH2 cells are generated from Ag-driven
differentiation of TH0 cells
 TH1 cells
 Synthesize IL-2, IFN-g and TNF-b
 Activate cells involved in CMI
 CD8+ T cells, NK cells and macrophages
 TH2 cells
 Synthesize IL-4, IL-5, IL-10 and IL-13
 Trigger B cells to class switch to IgE production
 Activate eosinophils
Th0, Th1, Th2
 Activation of CD4+ Cells by Ag
 Many Ags give rise to all three subsets
 Some Ags produce more of one subset than the other
 Viruses and bacteria favor the production of TH1 cells

 TH1 cells also associated with delayed-type hypersensitivity

 Allergens and parasites favor TH2 cell induction

 TH1 cells develop when IL-12, IFN- and IL-18 are present during Ag
stimulation
 These cytokines are products of innate immune components (NK

cells and macrophages)

 TH2 cells develop in the presence of IL-4
 Produced by mast cells or NK1.1 CD4+ T cells
Th1 vs. Th2 cells
 TH1 and TH2 can regulate each
other
 Cytokines produced by TH1 can inhibit function
of TH2 and vice versa
 IFN-g from TH1 cells inhibits generation of

TH2 cells
 IL-4 and IL-10 from TH2 cells inhibit TH1

cells
 Clinical Significance
 Regulating the balance of TH1 and TH2

subsets may be a way to treat different

diseases
 CD4+ T Memory Cells
 CD4+ memory T cells – long-lived cells that
carry out the secondary response to Ag
 Express a different form of CD45 (CD45RO)

 Less need for costimulatory interactions

 Different expression of certain CD

molecules
T Cytotoxic Cell Surface Molecules
 Function of CD8+ T Cells
 Major function is to kill cells that have been infected
by pathogens, such as bacteria and viruses
 Also involved in killing transplanted foreign cells

during graft rejection and tumor cells

 Referred to as “Killer” or Cytotoxic T Lymphocytes
 Cell killed by a CTL is known as a “target”, which
can be a specialized APC such as a dendritic cell, or
any other cell in the body
 Recognize peptide + class I MHC
 Results in death of the cell presenting the foreign
peptide
 CD8+ T Cells
 Also synthesize cytokines
 IFN-g, which regulates certain viral and bacterial infections

 TNF-b, which plays a role in target cell killing

 Others synthesize IL-4 similar to TH2 CD4+ T cells

 CTLs must be activated before they kill their targets

 One major pathway is via the activation of virus specific

CD4+ T cells
 Activated CD4+ T cells produce IL-2

 IL-2 + virus infected cell  activates CD8+ T cells

 Other CD8+ T cell activation does not require CD4+ T cells

 Alternative pathways involve presentation of Ag to CD8+

T cells by APC s expressing high levels of MHC class I

 Killing of Target Cells by Cytotoxic T
Cells
 Cellular interactions involved in activating CD8+ T cells 
similar to CD4+ events
 Activated CTL initiates killing by attaching to the target cell via
its TCR
 Contact is enhanced by various adhesion molecules

 Killing occurs by two pathways

 Secretion of perforin + granzymes

 Pores in target cell contributes to eventual cell death

 Granzymes (set of serine proteases)  induces apoptosis

 Cell surface Fas-Fas Ligand interaction

 Interaction activates apoptosis of the target cell via a

sequential activation of proteolytic enzymes known as

“caspases” inside the target cell
Fas
 Killing of Target Cells
 Activation of CTL and killing of the target cell are
separable events
 Can obtain virus-specific CTL from an individual that has
been infected with a virus
 These virus-specific CTLs are able to kill virus-infected
targets outside the body in vitro without addition of any
other factors
 killing of a target cell by a virus-specific CD8+ CTL
occurs via the recognition of a specific combination
of viral peptide associated with a particular MHC
class I molecule
Other Ways to Activate T Cells
 Superantigens (SAg)
 Class of Ag that activate T cells

expressing a specific V segment

 Bind only to V region of the TCR

 Presented by class II MHC but are not

bound in the peptide groove

 They are not believed to be processed

by APC
 Staphylococcus aureus and rabies

virus produce SAg

 Other Ways to Activate T Cells
 Plant proteins and Abs to T cell surface molecules
can activate many if not all clones of T lymphocytes
 These substances are referred to as polyclonal
activators or mitogens
 Ability to induce mitosis
 Plant glycoproteins (lectins)
 Concanavalin A (Con A) and phytohemagglutinin (PHA) are
particularly potent mitogens for T cells
 Pokeweed mitogens activates both T and B cells
 Bacterial LPS – mitogenic for mouse B cells
 Anti-CD3 Abs induce all T cells to proliferate
 T and B Cell Cooperation
 The generation of Ab in response to the vast
majority of proteins requires
 Ag + T cells + B cells
 Ags that require CD4+ T cells (helper T cells) are
referred to as thymus-dependent (TD) Ags
 Direct activation of B cells requires cross-linking of
the BCR
 Cross-linking Ags have multiple repeating epitopes
 Most proteins do not have multiple repeating units
and are thus TD Ags requiring CD4+ T cells for
production of Ab
 T and B Cell Cooperation
 The production of Ab in response to a TD Ag requires
that both T and B cells be activated and interact
 For T-B cooperation to occur, the T and B cells must
respond to epitopes that are physically linked in the
same Ag
 The B and T cells may respond to different epitopes
on the Ag, but these epitopes must be physically
linked in order to produce Ab
 Phenomenon of T-B cell cooperativity is also known
as “linked recognition”
 T and B Cell Cooperation
 Primary response T cells are most effectively activated by Ag
processed and presented by dendritic cells
 Activated T cells are then likely to interact with and activate B
cells in a fashion similar to the secondary response
 In secondary responses, both the relevant T and B cells have
been previously activated and expanded by Ag, and in contrast
to primary responses, very efficient T-B cell cooperation occurs,
with no requirement for a dendritic cell
 B cells internalize Ag + Ig  Process Ag  Present peptides +
class II MHC
 B cells acts as the APC in the secondary response
 To achieve linked recognition, it is important that the B cell
epitope and the T cell epitope be the same in both primary and
secondary responses.
T dep Ag
 Class Switching
 Class or isotype switching ti IgG, IgA or IgE requires
the interaction of CD40 on the B cell with its CD40L
on the T cell
 The cytokines produced by the T cell determines the
isotype of Ab synthesized by the switched B cell
 Highlights the importance of T-B cell cooperation for
a secondary response
 T-Independent Responses
 B cell Activation in the absence of T cell help can
occur with Ags (such as polysaccharides) that have
many repeating, identical epitopes on each
molecule
 Components of bacterial cell walls (LPS)
 Occurs through extensive cross-linking of BCR
 Called T-Independent (TI) responses
 Receptor cross-linking brings together more than
one BCR complex in the B cell membrane
 Cross-linking activates a cascade inside the B cell
similar to the events needs to activate CD4+ T cells
 TI Responses
 Two biologically relevant features if TI responses
 TI Ags generate primarily IgM and they do not give rise to
memory
 2nd exposure 
 same level of IgM production as the primary response
 No increase in level or speed of onset
 No class switch
 A protective response can still be made against TI Ags even
if an individual lacks T cells
 Patients with T cell immune deficiencies
 can still make protective IgM responses a/g extracellular bacterial
proteins
 Cannot make significan t
 The TCR Complex
 TCR is expressed on the surface of T cells in noncovalent
association with a complex of transmembrane polypeptides
 CD3 – 3 distinct polypeptide chains, ,  and

 Members of the Ig superfamily

 The  chain associates with both  and 

 “chaperone” role in transporting newly synthesized TCR

molecules to the cell surface

 Expressed exclusively on T cells

 Two identical  (zeta) chains

 16 kDa

 Found on T cells, macrophages and NK cells

 Mice also can have a(eta) form

 TCR Complex
 CD3 and  polypeptides
 Do not bind Ag

 When a + b bind Ag  change in CD3 and  that transfers

signal into the T cell

 Change in gene expression in the nucleus

 Crucial role as transducer molecules

 Both contain immunoreceptor tyrosine-based activation motif

(ITAM)
 Docking sites for protein kinases that activate the T cell
 CD4 and CD8 Molecules
 Transmebrane molecules
 Referred to as co-receptor or accessory molecules
 Member of the Ig superfamily
 T cells are either
 CD4  CD4(+)CD8(-)
 CD8  CD4(-)CD8(+)
 Immature T cells in the thymus can be CD4+CD8+
 In blood and tissue
 CD4+ to CD8+ ratio is slightly >2 to 1
 CD4+ constitute about 50-60% of total blood lymphocytes
 CD8+ about 20-25%
 CD4 and CD8 Functions
 Extracellular portion bind to the invariant portion of
the MHC on the APC  adhesion molecule
 MHC restriction
 CD4  binds Class II MHC
 CD8  binds Class I MHC
 Act as signal transducers
 Linked to specific kinases which are activated after
binding of MHC + peptide
 CD4 molecule binds HIV  allowing for virus entry
Interaction of the TCR with MHC
 The CDRs of the TCR V regions bind
both peptide and MHC
 The  T cells
 Some T cells express a TCR distinct from 
 Associated in association with CD3 and 
 NOT to be confused with the  and  chains of the
CD3
 Generally  cells lack CD4
 Some  do express CD8
 In normal adults found in much lower
numbers than  cells
 Numbers are increased by infectious agents
 The  Tcells
 The  and  lineages diverge early in intrathymic development
 Individual  cells appear in the thymus before  bearing cells
 Two subpopulations
 Skin or epithelial areas such as ling and intestine

 Function not well understood

 Do not respond to a wide range of protein Ags
 Some have been found to be activated by mycobacterial Ags and heat
shock proteins
 Synthesize cytokines in response to Ag
 May provide a first line of defense against invading pathogens
The  T cells
 Do not respond to complexes of peptide + MHC
 Some can recognize native Ag (such as viral
proteins)
 Some interact with other cell surface
structures
 Ag binding may be more like Ag binding to Ab
than to  TCRs
Genes coding for TCRs
 The  and  chains are constructed from V and J gene segments (like
light chains of Ab)
 The  and  chains are constructed from V, D and J genes
 The  and  genes are found on different chromosomes
 Gene segments of  and  loci are interspersed on the same
chromosome
 The mechanisms fro generating TCR diversity are very similar to the
mechanisms of generating diversity of the BCR
 Multiple germ line V genes
 Random combination of chains
 *Junctional and insertional variability
 NO somatic hypermutation after Ag stimulation
 Allelic exclusion
 Repertoire of different TCRs is believed to be as large or larger than Ig
(1015 –1018)
T Cell Differentiation in the Thymus
 Thymus (primary lymphoid organ) is
absolutely required for differentiation into T
cells
 Removal of thymus shortly after birth (mice)
 drastically deplete T cells
 Removal later in development  less impact
 Differentiation of T cells in the thymus
 MHC restriction of T cells
 T cells are made self-tolerant
 Thymocytes Interact with Thymic
Nonlymphoid cells
 T cell differentiation in the thymus is a complex multi-step
process
 At every stage of thymic maturation, developing T lymphocytes
(thymocytes) are in contact with a mesh of nonlymphoid
(stromal) cells of the thymus
 Trickle through this mesh from top (cortex)  medulla
 Nonlymphoid cells provide critical cell surface interactions for
development
 Also produce IL-7
 Most important thymic nonlymphoid cells
 Cortical epithelial cells

 Interdigitating dendritic cells (IDC)

 Found predominantly at the junction of cortex and medulla

 TCR Gene Rearrangement
 Early stages of differentiation in cortex  TCR  and  chain
genes start to rearrange more or less simultaneously
 If  chain rearrangement is successful  then d also start to

rearrange
 If both  and d rearrange “productively”   T cells

 If  and/or  rearrangements are not functional   gene

rearrangement continues
 If productive  rearrangement the  polypeptide is

expressed on the surface with an invariant molecule – pT

(or gp33) + CD3 = pre-T-cell receptor
 As a consequence of signaling through the pre-TCR further

rearrangement of  genes stops

 Ensures only one type of  chain expressed (allelic

exclusion)
 TCR Gene Rearrangement
 The  lineage first expresses both CD4 and CD8 on its
surface (CD4+CD8+)
 These cells are found in the cortex and forms the
majority of thymocytes in the young mammalian
thymus
 These cells express TCRs with specificities for all Ags
 both foreign and self
 These CD4+CD8+ T cells then undergo thymic
selection
 Thymic Selection
 1st stage “positive selection”
 TCR of the CD4+CD8+ T cell interacts with MHC on
epithelial cells in the cortex
 No interaction with thymic epithelial cells  apoptosis
 Positive selection results in the proliferation and
expansion of the CD4+CD8+ T cell and terminates any
further gene rearrangement
 As a consequence of this positive selection the  T cell
becomes “educated” to the MHC expressed on the thymic
cortical epithelial cells
 From this point on will recognize Ag bound to the type of
MHC that the developing T cell encountered in the thymus
 MHC restriction of T cell responses
 Thymic Selection
To prevent autoreactive T cells from leaving the
thymus  negative selection (2nd selection step)
 Interaction with interdigitating dendritic cells (IDCs) at
the corticomedullary junction
 TCR on the CD4+CD8+ T cell interact with MHC class
I and class II + self peptide presented on the IDC
 Presentation of “self” peptides result in high affinity
interactions  deleted by apoptosis
 Presentation of non-self peptides result in low affinity
interactions  survive to constitute the body’s CMI
 Thymic Selection
 Also as a consequence of the interaction with the IDC
 thymocytes that survive negative selection, down
regulate expression of CD4 or CD8
 CD4+CD8- or CD8+CD4- T cells leave the thymus to
circulate through the body  these are the
peripheral CD4+ or CD8+ T cells
 These CD4+ and CD8+ cells are
 MHC restricted
 Self tolerant
 Lymphocyte Trafficking
 Mature lymphocytes leave the thymus and circulate
via the blood to secondary lymphoid organs
 To leave circulation  involves multiple adhesive
interactions b/w circulating leukocytes and
endothelial cells at the boundaries of tissues
 Different lymphocytes preferentially enter different
tissues (homing)
 Naïve lymphocytes home to peripheral and mucosal
lymph nodes
 Activated and memory T cells move out of the nodes
and to sites in the skin and other tissues
adhesion
adh2
Interleukin-Induced Proliferation and Differentiation of an
Activated T4-Lymphocyte
Proliferation of a T4-Lymphocyte after
Activation
 An MHC- II Molecule on an Antigen-Presenting Cell
Presenting a Bound Viral Epitope to a T4-Lymphocyte with a
Matching TCR/CD4 on its Surface
An MHC- II Molecule on a B-Lymphocyte Presenting a Bound
Viral Epitope to an Activated T4-Lymphocyte with a Matching
TCR/CD4 on its Surface