Use the action buttons

to navigate through the tutorial

Back to previous slide Go to tutorial for

information

Forward to next slide Back to previous question

Back to beginning of tutorial

Forward to next question

Forward to last slide viewed

Answer selection

Back to case description

BEGIN HERE
 Effector T Cells
Secondary Lymphoid Organs
and Lymphocyte Trafficking
Jose I. Diaz, M.D., Ph.D.
jose.diaz@med.fsu.edu
 T Cells at Work or
Effector T Cells

Activated T cells are ready to work and

become effector cells
Effector CTL have one job, to kill cells
infected with virus, bacteria or
parasites
Effector Th cells have two jobs: 1)
travel from node to node to help B cells
becoming activated and CTL to expand
and 2) exit blood and travel to tissues
to help macrophages, B cells and T cells
 T Cells at Work or
Effector T Cells

Th cells are the quarterbacks of the

immune system, they mediate
communication of the immune system
cells very precisely by secretion of
cytokines such as -IFN, TNF, IL4,
IL5, IL6, IL10, IL17, IL21
These cytokines customize response to
any particular microbe
There are three major subsets of Th
cells identified so far: Th1, Th2 and
Th17
 Dendritic Cells and Th Cells
How Th cells know which cytokine
profile is the most appropriate?
Knowing this implies knowing: 1) type of
microbe and 2) body location
Dendritic cells answer these two
questions
PAMP receptors on dendritic cells
provide information about the type of
microbe; cytokines provides a sense of
identity about which part of the body is
under attack and also about type of
microbe
 Dendritic Cells and Th Cells

How do dendritic cell pass the defense

plan to Th cells?
co-stimulatory molecules and cytokines
produced by dendritic cells convey the
game plan to the Th cells
B7 is one of the best known co-activators
but there are many others, the type of
second signal used depend on the type
of microbe
Activated dendritic cells also produce
cytokines depending on the identity of
the microbe and the location of the
fight
 Th1 Cells

When dendritic cells and macrophages

recognize bacteria or viruses, macrophages
produce certain cytokines and dendritic cells
travel to nodes to present Ag and ! IL-12
IL-12 !Th cell ! Th1 cell ! TNF and
-INF ! activates macrophage and NK
cell and ! B cell Ig class switch to IgG3
(best opsonizer); Th1 cell ! IL-2 !
reactivates NK cell
This customized cytokine package is the
most effective against viruses or bacteria
Th1 Cells
 Th2 Cells

When dendritic cells and macrophages

recognize parasite or intestinal pathogenic
bacteria, activated dendritic cells travelling to
local nodes or MALT present Ag to nave Th
cells and program ! Th2 ! IL-4, IL-5 and
IL-13
IL-4 is a GF for Th2 cells and expands the
clone; IL-4 and IL-5 ! B cells to switch to
IgE (good parasite killer) or IgA (good
intestinal bacterial killer) depending on
location; source of initial IL-4 is unknown
Th2 Cells
 Th17 Cells
Th17 subset is a recent discovery and plays
a major role in the fight to fungi and
certain types of extracellular bacteria
When attacked by fungi, activated dendritic
cells travel to local nodes !TGF- and IL-
6 and co-stimulators commanding Th cells !
Th17 ! IL-17 and IL-21
IL-21 is a GF for Th17 cells and an
excellent recruiter of neutrophils, the best
defender against fungi and some
extracellular bacteria to whom Th1 and Th2
can be ineffective
Th17 Cells
 Th0 Cells

Some Th cells remain unbiased after

activation and dont become Th1 or Th2
cells but retain their ability to produce a
wide range of cytokines
These Th0 cells are instructed where to go
but not what to do; if they encounter an
environment rich in IL-12 ! Th1 cells, if
not some other type depending on the
cytokines
Th0 cells may become Th1, Th2 or Th17
cells on demand at the battle field
Th0 Cells
 Th Cell Subset Balance

Th1, Th2 or Th17 retain their cytokine

profile and produce GF giving them
positive feedback
There is also negative feedback to
opposites; Th1 cells IFN- inhibits Th2
cells and vice versa
Cytokines act locally, which explains why
immune systems can defend the host
simultaneously against different microbes
in different locations
 Th Cell Subset Balance

Th1/Th2 cell balance may

determine the outcome of some
infectious diseases, such as
Leprosy or TB
It is important to maintain and
adequate healthy balance
between the various Th Cell
Subsets
 Dendritic Cell and Th Cell
Summary
Dendritic cells collect information about the
microbial PAMP identified and the cytokines
produced by macrophages at the infection
site, integrate this information and generate
an adequate defensive plan; acting like the
coach of the Th cells
Dendritic cells travel to local lymph nodes,
use various combinations of co-stimulators
and cytokines and instruct Th cells: 1)
where the battle is taking place and 2) what
cytokines to produce in response to a
particular pathogen
 Dendritic Cell and Th Cell
Summary

Once Th cells are committed to a particular

cytokine profile, they secrete GF
stimulating their own proliferation; Th1
cells secrete -IFN, Th2 cells secrete
IL-4, and Th17 cells secrete IL-21
Committed Th cells then either 1)
circulate in the blood and lymphatic fluid
to provide help to B cells or 2) exit
blood at sites of infection and join the
battle helping macrophages or other
phagocytes
 Dendritic Cell and Th Cell
Summary

Uncommitted Th0 cells are also delivered

to the battle scene where under the
influence of battle cytokines become the
Th cell subset most wonted; secrete a
cytokine profile appropriate to the local
scene and the encountered microbe
Once a Th cytokine profile is
established, positive and negative
feedback tend to lock in this profile
 Question
Based on what you have learned on previous slides about CD4+
Helper Lymphocytes (Th cells), which of the following
statements is most correct?
Select the best statement

Th2 cells differentiate from Th0 cells under the influence of

TGF- and secrete IL-21, which help to fight fungal infections

Th1 cells differentiate from Th0 cells under the influence of

IL-12 and secrete -IFN, TNF and IL-2, which help to fight
bacterial and viral infections
Th17 cells differentiate from Th0 cells under the influence of
unknown cytokines and secrete IL-4 and IL-5, which help to
fight parasitic infections
Th0 cells is the most important subset of Th cells; they
coach dendritic cells and determine which cytokines they
should release
 Incorrect Answer!

Go back to the previous

question by selecting the
action button below and try
again.
 Correct Answer!
Th1 cells differentiate from Th0 cells under the influence
of IL-12 secreted by dendritic cells in response to most
bacteria and viruses. They secrete -IFN and TNF, which
enhance the functions of macrophages and B cells and IL-
2, which reactivate NK cells
Th2 cells do not develop under the influence of TGF-
and secrete IL-4 and IL-5 which command B cell Ig class
switch to IgE, which activates mast cells and eosinophils,
which can kill parasites; also may command switch to IgA,
which is excellent to fight GI bacterial infections
Th17 cells differentiate from Th0 cells under the
influence of TGF- and IL-6 secreted by dendritic cells
and produce IL-17 and IL21, which recruit neutrophils,
good fighters against fungi and pyogenic bacteria
Th0 cells is an important subset of Th cells that can adapt
to produce cytokines on demand; Th0 cells do not coach
dendritic cells; dendritic cells coach all Th cell subsets
 Effector CTL Killing

Activated CTL proliferates, leave the node,

enter blood and travel to tissues to kill
infected cells by cell mediated contact
Produces perforin, which drill holes on
infected cells membrane and inject
granzyme B !activates caspases !
apoptosis (same mechanism as NK cells),
killing exclusively infected cells
CTL ! FasL ! Fas on surface of target
cells ! apoptosis
 Effector CTL Killing

Cells that die by necrosis release lysosomes

and other contents damaging surrounding
cells and tissues
Cells that die by apoptosis release contents
enclosed in vesicles, so called apoptotic
bodies, which are cleared by the
macrophages garbage collector function
As virus infected cell dies on apoptosis, all
cell and viral DNA/RNA is equally destroyed;
watch the figure on next slide
 Effector CTL Killing
By inducing apoptosis on infected cells, CTL get
rid of virus infected cells without causing much
collateral damage to other cells, acting as a very
selective killer
 Secondary Lymphoid Organs
Immune Response Phases:
1) Recognition of danger
2) Production of weapons appropriate to the
attack
(type microbe, magnitude and site of attack)
3) Transport of weapons to site of attack
Recognition takes place in secondary lymphoid
organs:
Lymph Nodes
MALT (mucosal associated lymphoid tissue)
Spleen
Primary or central lymphoid organs such as the
Bone Marrow (BM) produce the immune cells; the
 The Logic of Secondary
Lymphoid Organs
Secondary lymphoid organs are strategically
placed to intercept microbes entering the
body at various locations and routes;
examples
Skin to local lymph nodes
Intestine to Peyers Patches
Blood to Spleen
They mobilize the most appropriate weapons
for the kind of microbes in those locations
Cytokine environments in various secondary
lymphoid organs determine the local
characteristics of the immune response
 The Logic of Lymph Nodes

Lymph nodes are lymph filters

detecting Ag from infected tissues and
providing a focused environment with a
concentration effect
Nodes facilitate the encounter of Ag,
APC and lymphocytes, the activation of
Nave B and T cells B and the
restimulation of experienced B and T
cells
 Lymphocyte Trafficking
How do lymphocytes know if they should go
to secondary lymphoid organs or tissues?
500 billions lymphocytes circulate each day;
traffic is carefully orchestrated to maximize
chances to find the microbes
The traffic of nave lymphocytes is very
different from the traffic of experienced
lymphocytes
T cells are born in the BM and educated in
the thymus; they express adhesion
molecules on their surfaces which are
passports for their destination
 Lymphocyte Trafficking
Nave T cells express L-selectin which binds
to GlyCAM-1 on HEV of nodes (lymph node
passport); similarly express 47 which binds
to MadCAM-1 on HEV of Peyers patches
draining to mesenteric nodes
If they dont encounter Ag in T cell area they re-
enter blood via lymph or directly in spleen; it
takes 12 to 24 hours for a complete round
and after many rounds of the same, they die in
apoptosis in about 6 weeks if never encounter Ag
If they do find Ag, they become activated,
proliferate and yield experienced T cells
 Lymphocyte Trafficking
Experienced T cells acquire restricted
passports depending on where they have
become activated; during activation
expression of certain adhesion molecules
decreases while other increases
T cells activated in Peyers patches express
high levels of gut-specific 47 and low L-
selecting and tend to return to Peyers
patches where there is a great chance to be
restimulated
Experienced T cells must also have combat
passports
 Lymphocyte Trafficking
Experienced T cells acquire adhesion
molecules to roll, sniff, stop and exit
blood to enter inflamed tissues; same
system used by neutrophils
T cells activated in a mucosa express high
levels of integrin E7 that binds to
addressin in blood vessels of inflamed
mucosa; battling chemokines direct them
Nave T cells have passports to visit
secondary lymphoid organs but DO NOT
have passport to visit sites of
inflammation, increasing their chance
become activated
 Lymphocyte Trafficking
Experienced T cells have restricted
passports to return to the secondary
lymphoid organs where they were
activated, increasing their chance to be
restimulated
Experienced T cells have also passports to
exit blood at sites of inflammation, increasing
their chance to go to the battle field and
destroy or help destroying microbes
Experienced B cells are not so migratory as
experienced T cells and settle in secondary
lymphoid organs or BM, produce Ab and let
this doing the travelling
 Lymphocyte Traffic in Nodes
Incoming lymphatics bring lymph which flows
from marginal to medullary sinuses exiting
through outgoing lymphatics
Incoming arterioles bring blood exiting on
veins; high endothelial veins or HEV are in
paracortex, which is where B and T cells
leave blood to enter the node
B and T cells can also enter the node from
lymph while travelling from one node to
another but can only leave the node via
efferent lymphatics, eventually enter blood
from the lymphatic system
 Immune Cell
Topography in Nodes
Nodes are highly organized into specific
zones hosting APC, T cells and B cells
FDC produce chemokine CXCL13 attracting
nave B cells express receptors for this
chemokine to lymphoid follicles
Activated Th cells upregulate CCR7 border
receptor guiding them to edges of the follicle
to meet B cells, then lose CCR7 and upregulate
a germinal center receptor
Up and down regulation of chemokine
receptors and chemokine production controls
traffic of immune cells in the node
 Ag and Dendritic Cell
Trafficking in Nodes
Ag enters node escorted in dendritic
cells, opsonized or bound to Ig, or free
in lymph
A lymph node is a lymph filter and a dating
place for APC and lymphocytes to meet;
is also the center for lymphocyte
activation
As Th cells pass through paracortex, there
is a small chance they will encounter a
dendritic cell presenting his specific Ag
and become activated
 Th Cell Traffic in Nodes
Before immune system produce Abs there
are four requirements
Dendritic cell presents Ag to Th cells
Th cells with specific TCR recognize specific
Ag
Opsonized Ag is presented to B cells by FDC
B cells with specific BCR recognize native Ag
It takes few days for Th cells to build
their clone; activated Th cells leave the
node via lymph, re-enter other nodes
from afferent lymphatics or via HEV from
blood quickly recirculating throughout the
body
 Th Cell Traffic in Nodes
The probability that a Th cell recognizing a
specific Ag peptide arrives to the node at
the same time than the B cell with same Ag
specificity is very small
This is why activated Th cells proliferate
and recirculate to many nodes giving a
much better chance to find their partner
B cell for same Ag who needs help for
activation
When Th cells recirculate to other nodes
and reencounter Ag, they become
restimulated and provide help activating
more B cells
 CTL Traffic in Nodes

CTL are activated in the paracortex

upon encountering specific Ag
presented by dendritic cells
Many activated CTL migrate to
inflamed tissues to kill infected
cells
Others re-enter other nodes and
recirculate
 Ag, Dendritic cell
and B and T cell Trafficking
Watch the animation on the next two slides and
identify the following:
Ag escorted by dendritic cells
Ag drained by lymphatic fluid
Migration of nave T cells from blood into
lymph nodes trough HEV; Th cell to B cell
zones and CTL to paracortex
Activation of CTL and Th cells and migration
of activated CTL and Th cells to other nodes
and infected tissues; recirculation of nave
CTL and nave Th cells to other nodes
If you need to watch this several times click back to
previous slide
C
D
4
8
CC
DD
84
 Lymph Nodes Swell
during Infection

Why nodes draining a site of infection swell?

This is due to:
1) proliferation and increase number of
lymphocytes
2) arrival and increased number of
macrophages to medullary sinus, which are
recruited by cytokines
3) retention of fluid due to inflammation
When microbe is eradicated, there is no enough
Ag to maintain activated lymphocytes and B and
T cells die from lack of Ag stimulation; swelling
goes away
 Question
Based on what you have learned on previous slides about T cell
trafficking, which of the following statements is LEAST correct?

Select the worst statement

Lymphatic fluid and free Ag arrives to the nodes from afferent
lymphatics, drain to subscapular and medullary sinuses and exit
the node though efferent lymphatics

Lymph nodes are lymph filters that concentrate Ag from

infected tissues facilitating their encounter with lymphocytes

Nave CTL and Th cells enter the nodes from blood through
capillaries, migrate to paracortex and if they dont recognize Ag
continue their journey to other nodes and tissues in search of Ag

Activated Th cells leave the nodes from efferent lymphatics,

travel to other nodes where they get re-stimulated and co-
activate B cells with same Ag specificity and eventually
reach the infected tissues
 Incorrect Answer!

Go back to the previous

question by selecting the
action button below and try
again.
 Correct Answer!
Nave CTL and Th cells enter the nodes from blood
through HEV, migrate to paracortex and if they dont
recognize Ag continue their journey to other nodes
and search of Ag. However, nave lymphocytes in
general only circulate through secondary lymphoid
organs and do not migrate to tissues unless they
become activated (there might be some exceptions to
this rule)