Ligand and cell signaling

pathways
Muhammad Shoaib Momen
MBBS, FCPS (Medicine), MD (Rheumatology)
BSMMU
 Overview of cell signaling
• Cells typically communicate using chemical signals
• These chemical signals- proteins or other molecules produced by
a sending cell, often secreted from the cell and released into the
extracellular space
• They can float over to neighboring cells
• Just as a journey of a thousand miles begins with a single step, so a
complex signaling pathway inside of a cell begins with a single key
event – the binding of a signaling molecule, or ligand, to its receiving
molecule, or receptor
 Overview of cell signaling
• Not all cells can “hear” a particular chemical message
• In order to detect a signal (that is, to be a target cell), a neighbor cell
must have the right receptor for that signal
• When a signaling molecule binds to its receptor, it alters the shape or
activity of the receptor, triggering a change inside of the cell
• Signaling molecules are often called ligands, a general term for
molecules that bind specifically to other molecules (such as receptors)
Ligand binding to receptor
Cell-surface receptors
• Cell-surface receptors are membrane-anchored proteins that bind to
ligands on the outside surface of the cell
• In this type of signaling, the ligand does not need to cross the plasma
membrane
• A typical cell-surface receptor has three different domains, or protein
regions:
A extracellular ("outside of cell") ligand-binding domain
A hydrophobic domain extending through the membrane
An intracellular ("inside of cell") domain, which often transmits a signal
• There are many kinds of cell-surface receptors, but here we’ll look at
three common types:
Ligand-gated ion channels
G protein-coupled receptors
Receptor tyrosine kinases
Ligand-gated ion channels
• Ligand-gated ion channels are ion channels that can open in response
to the binding of a ligand
• To form a channel, this type of cell-surface receptor has a membrane-
spanning region with a hydrophilic (water-loving) channel through the
middle of it
• The channel lets ions to cross the membrane without having to touch
the hydrophobic core of the phospholipid bilayer
Ligand-gated ion channels
• When a ligand binds to the extracellular region of the channel, the
protein’s structure changes in such a way that ions of a particular
type, such as Ca2+ or Cl− can pass through
• Changes in ion levels inside the cell can change the activity of other
molecules, such as ion-binding enzymes and voltage-sensitive
channels, to produce a response
• Neurons, or nerve cells, have ligand-gated channels that are bound by
neurotransmitters
Ligand-gated ion channels
Receptor tyrosine kinases
• Enzyme-linked receptors are cell-surface receptors with intracellular
domains that are associated with an enzyme
• In some cases, the intracellular domain of the receptor actually is an
enzyme that can catalyze a reaction
• Other enzyme-linked receptors have an intracellular domain that
interacts with an enzyme
Receptor tyrosine kinases
• Receptor tyrosine kinases (RTKs) are a class of enzyme-linked
receptors found in humans and many other species
• A kinase is just a name for an enzyme that transfers phosphate groups
to a protein or other target, and a receptor tyrosine kinase transfers
phosphate groups specifically to the amino acid tyrosine
Receptor tyrosine kinases
• Signaling molecules first bind to the extracellular domains of two
nearby receptor tyrosine kinases
• The two neighboring receptors then come together, or dimerize
• The receptors then attach phosphates to tyrosines in each others'
intracellular domains
• The phosphorylated tyrosine can transmit the signal to other
molecules in the cell
Receptor tyrosine kinases
• Receptor tyrosine kinases are crucial to many signaling processes in humans
• For instance, they bind to growth factors, signaling molecules that promote
cell division and survival
• Growth factors include platelet-derived growth factor (PDGF), which
participates in wound healing, and nerve growth factor (NGF), which must
be continually supplied to certain types of neurons to keep them alive
• Because of their role in growth factor signaling, receptor tyrosine kinases
are essential in the body, but their activity must be kept in balance:
overactive growth factor receptors are associated with some types
of cancers
• Dimerization plays a critical role in the regulation of another family of 
transmembrane proteins, the receptor tyrosine kinases. Specifically, 
ligand binding to the extracellular domain allows the intracellular
kinase domain to dimerize and cross-phosphorylate at regulatory
sites, leading to activation of the intracellular kinase domain. 
• Dimerization leads to transphosphorylation of the tyrosine kinase
domains of the receptor with signaling via downstream intracellular
pathways leading to cell activation and proliferation.
Phosphorylation
• Proteins can be activated or inactivated in a variety of ways
• One of the most common tricks for altering protein activity is the
addition of a phosphate group to one or more sites on the protein, a
process called phosphorylation
• Phosphate groups can’t be attached to just any part of a protein
Phosphorylation
• Phosphate are linked to one of
the three amino acids that have
hydroxyl (-OH) groups in their
side chains: tyrosine, threonine,
and serine
• The transfer of the phosphate
group is catalyzed by an enzyme
called a kinase, and cells contain
many different kinases that
phosphorylate different targets
RECEPTORS THAT RECRUIT MOLECULES
WITH ENZYMATIC ACTIVITY
Cytokine Receptors

• Cytokines are soluble peptide regulators with profound

effects on immune homeostasis and autoinflammatory
disease
• Cytokine receptors are organized into four classes
• Class I receptors are
Receptors for interleukin (IL)-2,
IL-6, IL-7, IL-15, and also for hormones such as erythropoietin and
prolactin
Cytokine Receptors
Class II receptors include those for type I and II interferon (IFN), IL-10, IL-20, IL-22,
IL-26, IL-28, and IL-29
Tumor necrosis factor (TNF) receptors
IL-1 receptors can be considered analogous to Toll like receptors (TLRs)
• Whereas class I and II cytokine receptors use the Janus kinase (JAK)/signal
transducer and activator of transcription (STAT) signaling pathway, TNFRs use
adaptor molecules called TNFR-associated factors (TRAFs) to recruit complexes
with different functions
JAK/STAT Pathway

• The Janus kinase-signal transducer and activator of transcription

(JAK-STAT) pathway plays a major role in transferring of signals from
cell-membrane receptors to the nucleus 
JAK/STAT Pathway

• The JAK family includes four tyrosine kinases:

JAK1, JAK2, JAK3, and tyrosine kinase 2 (tyk2)
JAK1 and JAK2 are involved in growth and development,
hematopoiesis and inflammation
• JAK3 and Tyk2 are predominantly important in the immune response
• Upon phosphorylation, STATs undergo conformational changes and
detach from the receptor
JAK/STAT Pathway

• JAK-STAT pathway has a critical role in the fate of T helper cells (Th)
• CD4+ Th cells can differentiate into multiple effector subsets, including
Th1, Th2, Th17, regulatory T cells (Tregs) and so on
• Th1 cells involve in the elimination of intracellular pathogens and
their hallmark cytokine is IFN-γ (activating of cell mediated immunity) 
• Th2 cells are major players in immunity against parasites especially
helminths and also contribute to allergic reactions. Th2 cells produce
IL-4, IL-13 (stimulate B cells to produce IgE, persitaltism enhancement
and mucous production) and IL-5 (eosinophil recruitment and
development)
• Th17 cells eradicate extracellular bacteria (neutrophil enriched
inflammations) and participate in antifungal responses
• Treg cells regulate other T cell subsets and suppress potentially
pathological immune responses. Indeed, their function is tolerance
maintenance
Inhibitors of the JAK-STAT pathway
• Cytokine responses and the JAK-STAT pathway are
under tight control of molecules, including
Protein tyrosine phosphatases (PTPs)
Protein inhibitor of activated STATs (PIAS)
Suppressor of cytokine signaling (SOCS) proteins
Protein inhibitor of activated STATs (PIAS)
• PIAS inhibit the STATs by interfering with their DNA binding function
Suppressors of Cytokine Signaling (SOCSs)
• SOCS protein family are eight members, including SOCS1, SOCS2,
SOCS3, SOCS4, SOCS5, SOCS6, SOCS7 and cytokine-inducible SH2
domain protein (CIS or CISH)
• SOCS1-7 and cytokine-induced STAT inhibitors (CISs), which
block STAT activation
• SOCS3 acts mainly by binding to the JAK and the cytokine receptor,
which leads to inhibition of STAT3 activation
• Altogether, SOCS proteins preferentially regulate the termination of
JAK-STAT signaling process
Therapeutic approaches
• Most of cytokine receptors play their important roles in the initiation
and development of various immunological diseases through the JAK-
STAT pathway
• Monoclonal antibodies are effective in the regulation of cytokines and
their receptors extracellularly but intracellular signaling proteins can
also be targeted in the clinical settings
Tofacinitib
• Tofacitinib is effective for controlling both innate and adaptive
immunity
• JAK3 is affected more than JAK1 and JAK2 is the least affected JAK by
Tofacitinib
• Tofacitinib ameliorates murine lupus and improves vascular
dysfunction
• Therefore, it may be beneficial in SLE and its associated vascular
dysfunction
IL-7 receptor and signaling, common γ
chain (blue) and IL-7 receptor-α (green)
• Since JAK3 unlike the other JAKs
is limited to cytokines of the
common γ chain family;
therefore, Tofacitinib inhibits IL-
2, IL-4. IL-7, IL-9, IL-15 and IL-21
cytokines, which use common γ
chain receptor
Tofacinitib
• JAK3 only signals in combination with JAK1
• These cytokines are integral to lymphocyte activation, function, and
proliferation
• Hormone-like cytokines erythropoietin,
thrombopoietin, growth hormone, granulocyte-macrophage
colony-stimulating factor (GM-CSF), IL-3, and IL-5 all signal through JAK2
• IL-6, IL-10, IL-11, IL-19, IL-20, IL-22, and IFN-α, IFN-β, and IFN-γ signal through
JAK1
• Baricitinib preferentially inhibits JAK1/JAK 2
Tofacinitib
• Tofacinitib has substantial nanomolar inhibitory potency in enzymatic assays against all JAK family
kinases except TYK2,
but in cellular assays it shows functional specificity for JAK1
and JAK1/3 versus JAK2
• Most now consider tofacitinib to be a pan-JAK inhibitor
• Reduction in NK cells has been demonstrated with tofacitinib treatment.53 It has been postulated that
IL-6 inhibition, which signals through JAK1, may play a role in the
efficacy of the molecule as well as AEs such as neutropenia
and hyperlipidemia, which have been seen with tocilizumab, a monoclonal antibody to the IL-6 receptor
• Tofacitinib does not block IL-1 or
TNF signaling
Tumor Necrosis Factor Receptor Signaling

• The TNF superfamily consists of 19 ligands and 29 receptors, which

play varied roles in inflammation, apoptosis, and proliferation
• TNF members generally have proinflammatory functions mediated in
part through the nuclear factor-κB (NF-κB) pathway
• TNF binds to two distinct receptors: TNFR1 and TNFR2
Tumor Necrosis Factor (TNF) superfamily
Tumor Necrosis Factor Receptor Signaling
• Receptors of the TNF superfamily can be categorized based on the
presence or absence of an intracellular death domain (DD), of which
the D-containing receptors are ubiquitous in expression
• TNFR1 contains a DD and is expressed in virtually all cell types,
whereas TNFR2 is expressed mainly on immune cells, endothelial
cells, and nerve cells
Tumor Necrosis Factor Receptor Signaling
• TNF signaling induces several signaling pathways, including activation
of NF-κB, MAPK, and apoptosis
• Upon TNF binding to the TNFR1, the DD recruits the TNFR-associated
death domain (TRADD) protein, which subsequently recruits the
Fas-associated protein with death domain (FADD)
• TNF can activate NF-κB, leading to cell survival and proliferation
Tumor Necrosis Factor Receptor Signaling
• NF-κB activation leads to the production of pro-inflammatory
cytokines, such as IL-6, IL-18, chemokines, cyclooxygenase 2 (COX2),
and 5-lipoxygenase (5-LOX)
• TNF signaling in addition induces pro-inflammatory cytokines
such as IL-6, and extra-cellular matrix-degrading enzymes
known as matrix metalloproteinases (MMP), which function
in cellular activation and migration
Tumor Necrosis Factor Receptor Signaling
• Ligands that belong to tumor
necrosis factor (TNF) superfamily
are originally expressed on cell
surfaces
• Most of them are oligomerized
into trimers and later cleaved by
metalloproteases
Tumor Necrosis Factor Receptor Signaling
• On cell plasma membrane, the
binding between TNF ligands and
their receptors in the tumor necrosis
factor receptor (TNFR) superfamily
initiates multiple intracellular
signaling pathways
• The cytoplasmic region of TNFR after
ligand binding will recruit the
scaffold protein called TNF receptor
associated factor (TRAF), which will
further turn on the NF-κB signaling
pathway
Tumor Necrosis Factor Receptor Signaling
• NF-κB is an important transcription
factor in inflammation as an
activator of numerous pro-
inflammatory cytokines and
chemokines
• The red dashed arrow in the figure
indicates the cleavage of TNF ligands
into their soluble forms, while the
black arrows illustrate the
downstream signaling pathways
activated by TNF ligand-receptor
interactions
Anti TNF Drugs
An antibody molecule: The Fc region is responsible for the effector functions of the
antibody, and the Fab region forms the antigen-binding site. Within the variable
regions are small areas of hypervariability, which determine antigen specificity
Infliximab is a chimeric monoclonal antibody with a murine variable region (shown
in red) fused to a human Fcγ1 Ig. Adalimumab and golimumab are fully human
monoclonal antibodies. Certolizumab is a humanized Fab’ fragment bound to
polyethylene glycol (PEG) molecules. Etanercept is a TNF receptor– Fcγ1 fusion
protein. Potentially immunogenic areas within each antibody construct are
indicated in red
Tumor Necrosis Factor Receptor Signaling
• Tumor necrosis factor (TNF), interleukin (IL)-1, and IL-6 are secreted
by various cell types, although within the inflamed synovium it is
generally considered that the activated macrophages that are present
in the synovium are probably the predominant source of
inflammatory cytokines in patients with RA
• IL-1 and TNF activate the endothelium, which is important in
recruiting inflammatory cells into the joint
• They also directly activate osteoclasts that cause bony erosions and
damaged bone -- a hallmark of uncontrolled rheumatoid
inflammation
T Cell Receptor
Antigen presentation stimulates T cells to become
either "cytotoxic" CD8+ cells or "helper" CD4+ cells
Signal One
• T cells are generated in the Thymus and are programmed to be
specific for one particular foreign particle (antigen)
• Once they leave the thymus, they circulate throughout the body until
they recognise their antigen on the surface of antigen presenting
cells (APCs)
Signal Two
• In addition to TCR binding to antigen-loaded MHC, both helper T
cells and cytotoxic T cells require a number of secondary signals to
become activated and respond to the threat
• In the case of helper T cells, the first of these is provided by CD28
• This molecule on the T cell binds to one of two molecules on the APC
– B7.1 (CD80) or B7.2 (CD86) – and initiates T-cell proliferation
• This process leads to the production of many millions of T cells that
recognise the antigen
Signal Two
• In order to control the response, stimulation of CD28 by B7 induces
the production of CTLA-4 (cytotoxic T-lymphocyte-associated protein
4) (CD152)
• This molecule competes with CD28 for B7 and so reduces activation
signals to the T cell and winds down the immune response
• The molecules ICOS, 4-1BB and OX40 are found on the T-cell surface
and are stimulated by their respective ligands which are typically
found on APCs
• Unlike CD28 and the TCR, ICOS, OX40 and 4-1BB are not constitutively
expressed on T cells
Signal Two
• Likewise, their respective ligands are only expressed on APCs
following pathogen recognition
• This is important because it ensures T cells are only activated by APCs
which have encountered a pathogen and responded
• Interaction of the TCR with peptide-MHC in the absence of co-
stimulation switches the T cells off, so they do not respond
inappropriately
Signal three
• Once the T cell has received a specific antigen signal and a general
signal two, it receives more instructions in the form of cytokines
• These determine which type of responder the cell will become – in
the case of helper T cells, it will push them into Th1 type (cells
exposed to the cytokine IL-12), Th2 (IL-4), or IL-17 (IL-6, IL-23)
• Each one of these cells performs a specific task in the tissue and in
developing further immune responses
• The resulting cell population moves out to the site of the infection or
inflammation in order to deal with the pathogen
Signal Three
• T cells have another receptor, or coreceptor, on their surface that
binds to the MHC molecule and provides additional strength to the
bond between the T cell and the target cell
• Helper T cells display a coreceptor called CD4, which binds to class II
MHC molecules, and cytotoxic T cells have on their surfaces the
coreceptor CD8, which recognizes class I MHC molecules
• These accessory receptors add strength to the bond between the T
cell and the target cell
Signal Three
• The T-cell receptor is associated with a group of molecules called
the CD3 complex, or simply CD3, which is also necessary for T-cell
activation
• These molecules are agents that help transduce, or convert, the
extracellular binding of the antigen and receptor into internal cellular
signals; thus, they are called signal transducers
• Similar signal transducing molecules are associated with B-cell
receptors
Helper-T-cell activation
• Helper T cells do not directly kill infected cells, as cytotoxic T cells do
• Instead they help activate cytotoxic T cells and macrophages to attack
infected cells, or they stimulate B cells to secrete antibodies
• Helper T cells become activated by interacting with antigen-
presenting cells, such as macrophages
• Antigen-presenting cells ingest a microbe, partially degrade it, and
export fragments of the microbe—i.e., antigens—to the cell surface,
where they are presented in association with class II MHC molecules
Helper-T-cell activation
• A receptor on the surface of the helper T cell then binds to the MHC-
antigen complex
• Other signals are required to activate helper T cell, provided by two
ways:
Stimulation by a cytokine or
Through a costimulatory reaction between the signaling protein, B7,
found on the surface of the antigen-presenting cell, and the receptor
protein, CD28, on the surface of the helper T cell
Helper-T-cell activation
• Helper T cells are divided into two general subpopulations—TH1
and TH2 cells—that have significantly different chemistry and function
• TH1 cells primarily produce the cytokines gamma interferon, tumour
necrosis factor-beta, and interleukin-2 (IL-2)
• TH2 cells mainly synthesize the interleukins IL-4, IL-5, IL-6, IL-9, IL-10,
and IL-13
• The main role of the TH1 cells is to stimulate cell-mediated responses
(those involving cytotoxic T cells and macrophages), while TH2 cells
primarily assist in stimulating B cells to make antibodies
B Cell Receptor
The B cell receptor (BCR) is a transmembrane protein on the surface of a B cell. A B cell
receptor includes both CD79 and the immunoglobulin. The plasma membrane of a B cell is
indicated by the green phospholipids. The B cell receptor extends both outside the cell
(above the plasma membrane) and inside the cell (below the membrane)
B Cell Receptor
• The general structure of the B-
cell receptor includes
a membrane
bound immunoglobulin molecul
e and a signal transduction
region
• Disulfide bridges connect the
immunoglobulin isotype and the
signal transduction region
Schematic representation of the B cell receptor signaling pathways. Aggregation of the BCR quickly
activate Src family kinases, including Blk, LYN, and FYN and the SYK and BTK tyrosine kinases. As
such, the process catalyzes the formation of a ‘signalosome’ that consists of the aforementioned
tyrosine kinases, the BCR and the adaptor proteins, for instance, BLNK and CD19, as well as
signaling molecules, such as P13K, PLCy2, and VAV
Rituximab
• Biologic agents that can deplete B cells (eg, rituximab) or inhibit
factors that activate B cells (belimumab) are used for the treatment of
a range of rheumatic and other diseases
• In addition to their central role in antibody production, B cells present
antigen to T cells, activate T cells, and promote the production of
proinflammatory cytokines, including interleukin (IL)-1, 4, 6, 8, 10, and
12; tumor necrosis factor (TNF)-alpha; vascular endothelial growth
factor (VEGF), etc
Rituximab
• It eliminates CD20-positive B cells, induces complement mediated
cytotoxicity, and stimulates apoptosis but has little and/or nonspecific
effects on autoantibody titers
• It is probable that some combination of these effects account for its
activity in autoimmune disease
• CD20 may play a role in Ca2+ influx across plasma membranes,
maintaining intracellular Ca2+ concentration and allowing activation of
B cells
Rituximab
• Rituximab (RTX) targets the transmembrane protein CD20 molecule on the
surfaces of some but not all B cells (eg, pro-B cells, plasma cells)
• RTX by binding to CD20, that is expressed on pre-B and mature B
lymphocytes, leads to apoptosis of these cells with antibody- and
complement-dependent cytotoxicity
• This mechanism of action leads to a selective peripheral B cell depletion for
more than 24 weeks
• However, other niches of B cells (eg, those in the synovium) are variably
depleted
• Repopulation of peripheral B cells occurs after 6–9 months from RTX course
Rituximab mechanisms of action; the three major independent mechanisms are (1)
antibody dependent cellular cytotoxicity (ADCC), (2) complement mediated
cytotoxicity (CMC), and (3) apoptosis; subset panel illustrates a schematic view of
CD20 structure and rituximab
Antibody dependent cell mediated
cytotoxicity (ADCC)
• Antibody dependent cell mediated
cytotoxicity (ADCC) is a cooperative
response involvingbinding of
antibodies produced by B-cells to
antigens
• Recognition of the constant fragment
(Fc) by effector cells
• Release of damaging substances by
effector cells (NK cells, macrophage,
eosinophil, etc)
• Destruction of the pathogenic
organism, tumor cells
Rituximab
• RTX has different mechanisms of
action through
Activation of the complement cascade
which leads to a direct lyse B cells by
complement-mediated cytotoxicity
The recognition by both Fcγ receptors
and complement receptors 1 and 3 on
macrophages causes phagocytosis
Antibody-dependent cell-mediated
cytotoxicity and interaction with NK
cells via FcγRIII and complement
receptor 3
Co-stimulatory Receptors

• The activation of specialised immune cells from the adaptive immune

response (i.e. B cells and T cells) is controlled by their specific antigen
receptor but also by co-receptors
T cell co-receptors
• In order to be fully activated and respond to intruders or damaged cells, naïve
T cells (that haven’t contacted antigen) need to be activated by two signals
• The first one is from the T-cell receptor (TCR)
• As the TCR recognises a small part of the antigen (called peptide), the antigen
will be activated
• The second signal (co-stimulatory signal) is provided by a costimulatory
molecule
• The better characterised one is CD28, among others (ICOS, OX40, CD46…)
• Without receiving a second signal, the T cells remain mainly unresponsive,
becoming anergic cells, or die
T cell co-receptors
• The adaptive  immune response
starts when an antigen
presenting cell (APC) recognises a
danger signal (virus, bacteria,
damaged self…)
• This induces its maturation, the
cutting of the antigen into small
peptides for presentation in a
complex with MHC, as well as the
induction of ligands of
costimulatory molecules (step 1)
T cell co-receptors
• This allows the recognition of
the peptide by the TCR of the
specific T cell, and its activation
by costimulatory molecules
• As a result, T cells proliferate and
are fully activated (step 2)
• Activated T cells then express co-
inhibitory receptors (CTLA-4, PD-
1) (step 3), which will terminate
the T cell response (step 4)
B Cell Co-stimulation
• The specific B-cell receptor (BCR, an immunoglubulin molecule
attached to the membrane of the B cell) recognises the antigen in
native form, as opposed to the TCR that recognises antigenic peptides
• Costimulation of B cells is notably achieved by the molecule CD40
• CD40 ligand is expressed on T cells. As B cells can also present
peptides to T cells, there is a dialogue between T- and B cells that
allows continuous B-cell proliferation and optimal activation leading
to their differentiation into plasmocytes, which are mature B cells
producing high affinity antibodies against the original antigen.
Abatacept
• Abatacept is a T cell co-stimulation modulator approved for the treatment
of rheumatoid arthritis, JIA, psoriatic arthritis
• The cytokines secreted by activated T cells are thought to both initiate and
propagate the immunologically driven inflammation associated with
rheumatoid arthritis
• Cytotoxic T lymphocyte antigen (CTLA)4 is closely related to CD28 in
structure, can bind to B7 molecules but with much higher
avidity, and curbs the activation and proliferation of T
cells.
• Abatacept prevents CD28 from binding to its counter-receptor, CD80/CD86,
due to its higher affinity for CD80/CD86
• A biologic agent, abatacept, that targets a key pathway in T-cell costimulation
• CD28 and CTLA-4 (CD152) are important in the regulation of T-cell activation and function and
antigen-presenting cells, on which they interact with their ligands, CD80 and CD86 (also termed
B7-1 and B7-2)
• CD28 binding has been found to decrease the activity of regulatory T cells (T reg) and to increase
T effector cell activity, while CTLA-4 may, by contrast, ultimately activate T effector cells without
suppressing T reg activity. Thus, targeting of these pathways can impact immune function.
• Abatacept — Abatacept is a soluble fusion protein comprising CTLA-4 and the Fc portion of
immunoglobulin G1 (IgG1) (CTLA4-Ig). Abatacept is available for use in the treatment of
rheumatoid arthritis, juvenile idiopathic arthritis, and psoriatic arthritis
• Abatacept prevents CD28 from binding to its counter-receptor, CD80/CD86, due to its higher
affinity for CD80/CD86. This prevents the suppression of T reg activity and prevents increased T
effector cell activity
INNATE RECEPTOR SIGNALING
• Several components of the innate arm recognize what is foreign by
detecting certain carbohydrates or lipids on the surface of microorganisms
that are different from those on human cells
• Components of the innate arm have receptors called pattern-recognition
receptors that recognize a molecular pattern called a pathogen-associated
molecular pattern (PAMP) that is present on the surface of many
microbes but not present on human cells
• By using this strategy, these components of the innate arm do not have to
have a highly specific receptor for every different microbe but can still
distinguish between what is foreign and what is self
INNATE RECEPTOR SIGNALING
• There are two classes of receptors on the surface of cells
(Toll-like receptors and C-type lectin receptors) that recognize
microbes outside of cells and two classes of receptors in the
cytoplasm of cells (NOD receptors and RIG-I helicase receptors) that
recognize microbes that have invaded the cell’s cytoplasm within
cells
• Mutations in the genes encoding these pattern receptors result in a
failure to recognize the pathogen and predispose to severe bacterial,
viral, and fungal infections
Toll-Like Receptors
• The most important of these PRRs are the Toll-like receptors (TLRs)
• This is a family of 10 receptors found on the surface of many cells, including
epithelial cells and innate immune cells, such as macrophages and dendritic
cells
• Each of the ten TLRs recognizes a core microbial building block (e.g.,
endotoxin or peptidoglycan), and the resulting signal activates transcription
factors that enhance the synthesis of proinflammatory
cytokines and cell surface molecules
• The result is a rapid innate immune response, triggered by a particular
microbe in a particular location
Toll-Like Receptors

• Endotoxin is a lipopolysaccharide (LPS) found on the surface of most gram-negative

bacteria (but not on human cells)
• When released from the bacterial surface, LPS combines with LPS-binding protein, a
normal component of plasma, which transfers LPS to a receptor on the surface of
macrophages called CD14
• LPS then stimulates a PRR called Toll-like receptor 4 (TLR4), which transmits a signal
to the nucleus of the cell
• This induces the production of cytokines and surface proteins that are required to
activate helper T cells and to produce antibodies
• Excessive macrophage PRR activation is an important cause
of septic shock and death in hospitalized patients
Toll-Like Receptors
• Intracellular TLRs recognize nucleic acids derived from bacteria and
viruses, and also recognize self-nucleic acids in disease conditions
such as autoimmunity
• All TLRs are synthesized in the ER, traffic to the Golgi, and are
recruited to the cell surface or to intracellular compartments such as
endosomes
• Intracellular localization of TLRs is thought to be critical for ligand
recognition as well as for preventing TLRs from coming into contact
with self-nucleic acids, which could cause autoimmunity
Toll-Like Receptors
• Cellular location of TLRs and the
identity of their ligands/agonists
• The stimulation of surface TLRs
(TLR-2, TLR-4, and TLR-5) with
appropriate ligands results in the
activation of NFκB
• The ensuing increase in levels of
pro-inflammatory cytokines and
the influx of inflammatory cells
protects against both virus and
bacterial challenge
Toll-Like Receptors
• Activation of intracellular TLRs
(TLR-3, TLR-7, TLR-8, and TLR-9)
leads to IRF (Interferon
regulatory factor) activation and
the production of Type 1 IFNs
and pro-inflammatory cytokines,
again providing an environment
not conducive for pathogens
Toll-Like Receptors
• When activated, TLRs recruit adapter
molecules within the cytoplasm of cells
in order to propagate a signal
• Four adapter molecules are known to be
involved in signaling known as MyD88,
Tirap (also called Mal), Trif, and Tram
• The adapters activate other molecules
within the cell, including certain protein
kinases (IRAK1, IRAK4, TBK1, and IKKi)
that amplify the signal, and ultimately
lead to the induction or suppression of
genes that orchestrate the inflammatory
response
INTRA-CELLULAR RECEPTOR SIGNALING

• Intracellular receptors are receptor proteins found inside of the cell,

typically in the cytoplasm or nucleus
• In most cases, the ligands of intracellular receptors are small,
hydrophobic (water-hating) molecules, since they must be able to cross
the plasma membrane in order to reach their receptors
• For example, the primary receptors for hydrophobic steroid hormones
synthesized from cholesterol in the adrenals and gonads, and vitamin D
• When a hormone enters a cell and binds to its receptor, it causes the
receptor to change shape, allowing the receptor-hormone complex to
enter the nucleus and regulate gene activity and alters transcription
INTRA-CELLULAR RECEPTOR
SIGNALING
• Because they are lipid soluble, these hormones can diffuse through
plasma membranes to the nucleus to interact with receptors that
directly modulate transcription, making them potent and common
drug targets
• Almost half of the roughly 50 mammalian nuclear receptors have
poorly characterized or multiple low-affinity ligands and are therefore
known as orphan receptors
INTRA-CELLULAR RECEPTOR
SIGNALING
• Nuclear hormone receptors are broadly divided into two classes
• Type I are retained in the cytoplasm until ligand binding drives a
conformational change that releases binding partners, after which
they translocate to the nucleus
• Examples include the glucocorticoid, androgen, and estrogen receptors
• Glucocorticoid agonists, such as dexamethasone and prednisolone, are
important immunosuppressants
• Activation not only drives gene expression changes following nuclear
translocation but also transrepression of pro-inflammatory transcriptional
complexes, such as AP-1 and NF-κB
INTRA-CELLULAR RECEPTOR
SIGNALING
• Type II nuclear receptors generally reside on deoxyribonucleic acid and are inhibited
by co-repressor proteins until ligand detection
• Examples include thyroid hormone receptor (TR), retinoic acid receptor (RAR), and
vitamin D receptor (VDR)
• They act in conjunction with cor-receptors, the most common of which is the
retinoid X receptor (RXR)
• Although its major action is to regulate calcium absorption in the intestine and bone
remodeling, vitamin D has increasingly been recognized as
an immune modulator
• Macrophages within granulomas convert vitamin D into its active form as part of an
innate immune regulatory cascade
Conclusion
• Discovery of these signaling pathways has provided valuable
insights into immune pathogenesis of autoimmune and
immunodeficiency diseases
• Many receptors and/or signaling pathways are potential therapeutic
targets for rheumatologic diseases