B cells effector functions and tolerance – T cell assistance

B cells, antigens, and antibodies

B lymphocytes, alongside T lymphocytes, play a critical role in adaptive immune responses. B cells
originate from and complete most of their development in bone marrow (thus the name 'B' for
bone marrow), where they become immunocompetent (able to recognize a specific antigen) and
are continually produced even in adults. They then migrate to secondary lymphoid tissues such
as spleen, lymph nodes and tonsils where they will aggregate in the form of follicles until they are
called on during an infection when they expand and form germinal centers.
B cells recognize soluble (present in blood and lymph) pathogenic antigens via antibodies
(immunoglobulins) bound to their surface which destroy extracellular pathogens and their
products. This is known as humoral response which contrasts with cellular responses occurring
inside the infected cell and mediated by T cells.
B cell development and lineage commitment
Most B cells in the adult which reside in secondary lymphoid tissues originate from the
uncommitted stem cell in the bone marrow. There is also a small proportion of B cells which
derive from early fetal progenitors and usually reside in mucosal and epithelial tissues.
Look at the figure below to get familiar with different stages of B cell development and their
localization.
B cell functions
Main function is the production of antibodies in response to challenges (infections, vaccination).
Over the course of the immune response, the characteristics of the antibody response change.
When an individual is first exposed to an antigen, B cells replicate to make more of themselves
and produce an antibody response, mostly comprised of low affinity IgM type antibodies shown
here in this maroon, red color. As the primary immune response continues, the B cells switch to
making another type of antibody called IgG and that's shown here in green. As this response
resolves, the amount of antibody decreases but doesn't return to zero and you can see that here
in green. During this response, B cells develop into memory B cells that can live for a long time.
When an individual is exposed to the same antigen again, the memory B cells are recalled to the
response making a much stronger response and this is mostly composed of high affinity class
switched antibodies. The high affinity class switched antibodies bind to their targets very strongly
and are the most effective antibodies for neutralization. This type of antibody response is the
basis of most vaccine protocols.

B cells develop in the bone marrow, where they begin rearranging the VDJ segments of the
immunoglobulin genes, and this allows them to produce a functional B Cell Receptor. During this
developmental process, B cells go through several checkpoints which you can see in the image
below. At the immature stage where the B Cell Receptor is expressed, B cells are screened for
autoreactivity. High affinity interactions at this stage of development indicate order reactivity and
these cells undergo apoptosis. If the B Cell Receptor has no interaction at all, this indicates a non-
functional B Cell Receptor and the cells die by neglect, lack of a survival signal. Those B cells with
receptors that have low affinity for self-antigen go through the rest of the maturation process and
leave the bone marrow. This kind of low affinity signaling is called tonic signaling and this is
important for keeping naive B cells alive in the periphery. As mentioned, antibody production is
the main function of mature B cells. The antibody produced by B cell is essentially the same as its
B Cell Receptor where the antibody is the secreted form, and the B Cell Receptor is on the B cell
surface. The basic structure of an antibody is shown below. The antibody is comprised of four
chains, two heavy and two light chains. These chains come together to form a single structure
with two distinct regions. The variable region here in magenta contains the most diversity as this
is where the random combination of VD and J segments come together. The constant region here
in blue is conserved for specific antibody subtypes.
- B cell survival during development depends on the affinity of the interaction between
their B Cell Receptor and self-antigens

- The constant region is responsible for the downstream that dictate the antibody function
by binding to Fc receptors.
- The variable region is the part that binds to the antigen. It dictates antigen specificity. This
is why VDJ recombination is concentrated in this region.
Antibodies have several functions. They combined two targets and therefore block or neutralize
their function. In the case of viruses, this could prevent them establishing an infection. Antibodies
can also activate the classical pathway of the Complement cascade. A process that can lead to
license of bacteria via formation of the membrane attack complex. Antibodies can also activate
innate cells such as macrophages and natural killer cells via Fc receptors which are expressed by
these immune cells. This can potentiate their activation and promote effective functions such as
phagocytosis or killing.
The avidity concept is explained a little further here in the diagrams. On the far left, a single arm
of the antibody is binding to a single antigen. This is a monovalent interaction and has low avidity.
In the middle, an antibody binding to its specific antigen using both arms, so is bivalent and this
has slightly higher avidity. On the right-hand side, we have a pentameric display of antibodies,
which has very high avidity because there are 10 arms binding the antigen. Different classes of
antibodies have different capacity to make these polyvalent structures. The IgM class can create
this pentameric structure. So, the fact that it is low affinity, is compensated for by its high avidity.
The IgG class of antibody does not come together in a polyvalent structure and thus it relies on
its high affinity for strong binding to the antigen.

There are five main types of antibodies and each of these has their own unique characteristics
aside from their ability to form polyvalent or monovalent structures. IgM is well known because
of that pentameric structure, but it is also the first antibody produced during the primary immune
response. It is very good at activating complement and is also the B Cell Receptor through which
B cells are first activated. IgG is the main antibody switched to during an immune response and is
the main antibody type produced in secondary immune responses. It is monomeric and it tends
to be high affinity. Can activate complement very well and can also interact with immune cells via
Fc receptors. It's also very good at neutralizing toxins and viruses. IgA has a dimeric structure and
is strongly associated with mucosal tissues. It has an important function in maintaining the
balance of commencing bacteria. IgE is strongly associated with parasitic and helminth infections
where it is protective, but also with allergic responses such as asthma and atopic dermatitis,
where it is disease causing. IgD is expressed mainly by mature B cells that have not undergone
class switching. We don't really know what it does, but it may play a role in based on maturation
or homeostasis.
They each have their own effective functions, and this is mediated by the constant region. The
constant region binds to Fc receptors, which are specific for the constant region type and the
receptors themselves are expressed on a wide range of immune cells.

Let's start with Fc Gamma receptors. Fc Gamma receptors are specific for IgG antibodies, and they
can have different affinities for IgG subclasses:
- Fc Gamma receptor 1, also called CD64, is expressed on macrophages and has high affinity
for IgG, is important for promoting phagocytosis.
- Fc Gamma R3, also called CD16, is expressed by many innate immune cells. In the case of
NK cells, the recognition of IgG via NK cell expressed Fc gamma R3 allows for targeted
cytotoxicity, called antibody dependent cell-mediated cytotoxicity or ADCC. Not all Fc
receptors are activators though.
- Fc Gamma RIIB, which is expressed by B cells, is an important negative regulator of
antibody production. Mutation or loss of Fc Gamma RIIB is strongly associated with
increased risk of autoimmune diseases such as lupus.
There are two main receptors for IgE:
- The high-affinity Fc Epsilon receptor 1 is expressed mainly by basophils and mast cells and
additionally on eosinophils and monocytes in humans. Activation of Fc Epsilon R1 by
antibody cross-linking results in activation of Fc receptor bearing cells, typically causing
release of pro-inflammatory molecules.
- Fc Epsilon R2 has lower affinity for IgE than Fc Epsilon R1. It is also called CD23 and is
expressed on many immune cells and some stromal cells. On B cells, Fc Epsilon R2 is a
negative regulator of IgE production.
There are also two receptors for IgA:
- Fc Alpha receptor 1 promotes the activation of several immune cells. This includes
monocytes, neutrophils, and eosinophils. This enhances their phagocytic ability.
- The poly Ig receptor: The major function of IgA is to maintain the balance of commensal
bacteria. To do this, the IgA must reach the mucosal surface. This is achieved via the poly
Ig receptor. This receptor doesn't have any effect of function. Its only role is to actively
transport IgA across the barrier wall to the mucosal site. It is not the only antibody
transporter.
- The neonatal Fc receptor transports maternal IgG from the breast milk into the neonate,
as well as across the placenta into the fetus. The neonatal Fc receptor also regulates IgG
and serum albumin turnover, extending the half-life of these proteins by protecting them
from lysosomal destruction and allowing them to be recycled.
Not all Fc receptors are activating. Whether an Fc receptor's activating or inhibitory is
determined by the intracellular signaling cascades that stem from their cytoplasmic tails. This
is essentially controlled by immunoreceptor tyrosine motifs that associate with the Fc
receptors cytoplasmic tail. This is outlined in these diagrams. If you focus on the left image,
you can see the activating receptor Fc Gamma R1 in pink and the cytoplasmic tail is associated
with the immunoreceptor tyrosine activating motifs or ITAMs in green. When the receptor is
ligated, the ITAM motif is phosphorylated, and this leads to intracellular signaling. In the case
of inhibitory receptors such as Fc Gamma RIIB here in blue on the right-hand side. The
cytoplasmic tail is associated with immunoreceptor tyrosine inhibitory motifs or ITIMs also in
blue, which recruit phosphatases that essentially shut down the signaling process.

Fc receptors can also act via recruitment and interaction with immune cells. Typically, by cross-
linking cell surface Fc receptors. In antibody dependent cellular cytotoxicity-ADCC, antibodies
bind to antigens on the surface of a virally infected cell. NK cells recognize the bound antibody
via Fc receptors, leading to cross-linking of the Fc receptors. This leads to formation of an
immune synapse shown in the diagram. At the immune synapse, the directed release of lytic
enzymes occurs, allowing delivery of cytotoxic molecules such as perforin. This induces
apoptosis in the target cell.
A similar process occurs with cross-linking of Fc Epsilon receptors on mast cells, but with a
very different outcome. In this case, IgE is pre-bound to the Fc Epsilon receptor on the mast
cell. When an antigen comes along, for example, following pollen exposure in a sensitized
individual, the antigen binds to the IgE, which remember, is already bound to the Fc Epsilon
receptor. This cross-links the IgE Fc Epsilon receptor complex and that causes mast cell
degranulation. This degranulation releases lots of pro-inflammatory substances such as
histamine, which are a major cause of allergies such as hay fever.

It is also possible for the antibodies on the basal surface, the B cell receptor, to be cross-linked
by antigen binding. The signaling pathway for this is quite complex, but the result is the
activation of B cells and the phagocytosis of antigens. After they phagocytose antigen B-cells
can process this and present it by MHC Class II to CD4 positive T cells. By doing this, the B cell
receives help from CD4 T cells, and this help promotes the proliferation, survival, and
differentiation. The T cell of that provides help to B cells is a specialized subset of CD4 T cells
called follicular helper T cells. These cells are antigen-experienced, and they express the
chemokine receptor, CXCR5, which promotes the localization near the B cell area so they can
interact with B cells. Tfh express many co-stimulatory molecules and cytokines that together
promote B cell activation. A key co-stimulatory molecule is called CD40 ligand. CD40 is
signaling in B cells is crucial for their activation and isotype-class switching. Deficiency in CD40
ligand leads to high pyogenes syndrome, where class switching is impaired, and these
individuals are highly susceptible to infections.
When B cells recognize antigen, they become activated. This alongside co-stimulation and
cytokine help from T cells promotes B cell proliferation. As this process continues, B cells
ultimately differentiate into plasma cells or memory B cells. B cells need a bit of help to do all
these things, and this help comes from the Tfh cell. Tfh cells provide help to B cells at multiple
stages of the B cell response.

Here on the left, they promote B cell activation and class switching before B cells enter the
germinal center response. This happens at the border between the B cell follicle and the T cell
zone. After a successful interaction here, Tfh cells and B cells then move into the B cell follicle
to initiate the germinal center, shown in the middle. In the germinal center, Tfh cells provide
ongoing help to B cells while they undergo affinity maturation. On the right-hand side image
Tfh also promote the differentiation of B cells into memory B cells or plasma B cells. The
ultimate output of the germinal center response are these cell types. At each of the stages, B
cells produce antibodies that help fight infections.
Early on, plasma cells are made via the extra follicular pathway, which means they don't go
through the germinal center response. These plasma cells are short-lived, although they do
produce high amounts of low affinity antibodies, which is important to combat an ongoing
infection. B cells can also enter the germinal center response. The germinal center is a
transient cellular niche that allows the proliferation, somatic hypermutation, and selection of
B cells. Together, these processes lead to affinity maturation. The result is the production of
memory B cells and long-lived plasma cells that produce high affinity class switched
antibodies. This response can last a long time and essentially mediates protection from
secondary infection.
Evidence of the importance of B cells comes from people with deficiencies in B cell function.
This normally results from reduced antibody levels and increased susceptibility to infections.
Examples of this include hyper IgM syndrome, where class switching is impaired. There's also
a deficiency in recombination activating gene-RAG which results in poor or no B cell receptor
gene rearrangement. As well as splenectomy or removal of the spleen. B cells undergo part
of their maturation in the spleen, so when the spleen is removed B cells do not mature
properly and can have impaired responses to infection or vaccination. B cells can also
contribute to or cause diseases. In the case of allergy, B cells react to environmental antigens
that are not pathogenic. Examples of this include allergic rhinitis, atopic dermatitis, and
asthma. As you may recall, B cells are also the causative agent in many autoimmune diseases.
In these cases, antibodies are directed against tissue self-antigens. This leads to chronic auto
inflammatory responses that can be incredibly debilitating. There are many examples such as:
autoimmune hemolytic anemia, ANCA vasculitis, myasthenia gravis, and lupus. Each of these
diseases have auto-antibodies, auto-antigens that are targeted by antibodies and cause
symptoms and disease.
B Cell Tolerance
Tolerance is an important immunological process that removes potentially auto-reactive cells
from the immune system. This is a continual process. This takes place in two distinct phases
called central and peripheral tolerance:
- Central tolerance takes place in primary lymphoid tissue, the bone marrow for B cells, and
the thymus for T cells.
- Peripheral tolerance occurs outside these tissues, typically in secondary lymphoid organs.
Tolerance is an important process as it ensures that the immune cells do not respond to self-
antigens and thus limits autoimmune disease. Tolerance also helps limit reactions to
exogenous environmental antigens that are harmless. For example, food or airborne antigens
like pollen. From a therapeutic perspective, understanding tolerance will help us find a better
way to ensure acceptance of organ transplants in those individuals who need them. The ability
to break tolerance could also be of use in cancer where activating the immune response
against self-tissue would be clinically beneficial.

The stages of B cell tolerance can also be thought of in terms of tolerance checkpoints. There
are specific cellular and molecular interactions that must occur to determine B cell survival
and maturation. B cells develop in the bone marrow going through several stages of
development, each of which has important checkpoints. We estimate that around 90 percent
of B cells made in your bone marrow every day die before they're able to emerge from the
bone marrow into the periphery. This suggests that B cells are actively removed. Auto-reactive
B cells are recognized as such and deleted from the system before they can cause damage. In
the bone marrow, there are several mechanisms through which B cells can be tolerized.

The major mechanism is the deletion of self-reactive B cells by apoptosis, shown on the left.
In this case, self-antigens are displayed on bone marrow stromal cells. If a B cell receptor binds
to the self-antigen with high affinity, then the results in B cell receptors signaling leads to
inhibition of survival proteins such as Bcl2 triggering apoptosis, or the activation of cell death
pathway receptors such as Fas. B cells can also undergo a process called B cell receptor editing,
shown in the middle on the left, which changes the B cell receptor specificity. This can reduce
B cells affinity and can occur either through recombination of a second B cell receptor locus
or through hypermutation. The fate of a B cell is ultimately determined by BCR signal strength.
This can also be influenced by factors that are not directly linked to the BCR itself. These are
shown on the right-hand side images. For example, B cells can fine tune the level of BCR
signaling by altering expression of co-receptors or internal sibling components. This is what
we call B cell intrinsic regulation. B cell extrinsic factors can also influence B cell tolerance,
provision or limitation of co-stimulatory ligands or cell survival factors can influence the
induction of apoptosis and thus B cell survival in the bone marrow.
There are additional checkpoints on basal tolerance in the periphery. The germinal center is
one such stage where B cell receptor affinity is constantly checked and selected for as part of
affinity maturation. After activation, B cells will receive help from Tfh cells into the germinal
center. In the GC, B cells undergo somatic hypermutation to increase affinity for the antigen.
Because this is a random process, the BCR must be checked to ensure it can still bind the
antigen, and this occurs in a competitive environment. The checkpoint involves two other cell
types. The B cell interacts with follicular dendritic cells to capture antigen from them.
Follicular dendritic cells are a fibroblast-like mesenchymal cell that acts as a depot for antigen
in the germinal center.

The higher the affinity BCR for the antigen, the more likely the B cell will acquire it and acquire
more of that antigen. The B cell then internalizes the antigen and presents it to follicular
helper T cells. Tfh cells then provide help in the form of survival signals and co-stimulation. B
cells with high affinity BCRs will acquire more antigen and therefore present more antigen to
Tfh cells, so they will compete better for that T cell help and this ensures their survival. The
ultimate output of the germinal center is long-lived plasma cells and memory B cells. You can
see that the germinal center reaction can be considered a form of peripheral tolerance. The
combined action of hypermutation and selection leads to increase B cell receptor affinity, and
the B cells with higher affinity have a competitive advantage. They outcompete lower affinity
B cell receptors and receive more T cell help. You can appreciate again that this is a feed -
forward loop that improves affinity for foreign antigen. If this becomes dysregulated, however,
the GC process can promote autoimmunity. While germinal center is normally formed in
secondary lymphoid organs, they can also occur in peripheral tissue. This typically happens in
chronic inflammatory conditions and is particularly noted in autoimmune diseases and in
cancer. In both these settings, the present of ectopic germinal centers is associated with more
potent immune responses. In autoimmunity, this is detrimental and in cancer this is beneficial.
The development of ectopic germinal centers in these settings can be considered a form of
breaking tolerance. The idea is that when the germinal centers develop outside of secondary
lymphoid tissue, they are separated from the normal tolerance checkpoints that are present
in that secondary lymphoid tissue. This could lead to excessive exposure to self-antigens and
inflammatory mediators. That together will promote B cell activation and survival and
ultimately exacerbation of autoimmune disease via the breakdown of tolerance. It is
important to note that we don't really understand the etiology of ectopic germinal center
formation, although they are of great interest in chronic inflammatory diseases, B cell
depletion therapy is a well-recognized treatments for auto-immune diseases. By depleting B
cells, several things that are known to promote auto-immune disease severity are targeted.
Ectopic germinal centers are disrupted, limiting the ongoing breakdown of tolerance.
Regulatory T cells can be better induced, leading to reductions in inflammatory cytokines such
as tumor necrosis factor and induction of regulatory cytokines such as Interleukin 10. The
changes to cytokine profiles could be directly due to depletion of B cells. Activated B cells can
make high quantities of TNF. There is a unique subset of B cells called regulatory B cells that
make IL-10. In fact, in mouse models of multiple sclerosis, B cell derived IL-10 has been shown
to be essential for the suppression of the T cell driven autoimmune response. This highlights
the B cells do more than just make antibodies.
B cell responses to antigens
Mature B lymphocytes, equipped with B-cell receptor (BCR), also known as surface
immunoglobulin, migrate between secondary lymphoid organs (SLO) in pursuit of the one
type of antigen it binds to, its cognate antigen.On binding to a microbial antigen, B cells, which
also act as antigen presenting cells, receive help from T cells. These B cells can either
differentiate into IgM producing short-lived plasma cells or form the germinal center (GC). In
GCs, localized to SLO follicles, B cells undergo several processes which generate antibodies
that are more effective in eliminating pathogens. These processes include somatic
hypermutation (SHM), which mutates the genes used to produce antibodies, enabling affinity
maturation that 'fine-tunes' B cell specificity, by increasing their affinity to a particular antigen.
Selected B cells can also undergo class switching (e.g. from IgM to IgG antibody isotype),
which leads to the production of antibodies with different functions. Eventually, memory B
cells and plasma cells expressing high affinity BCRs with switched isotypes leave the germinal
center (GC).
T cell activation, T helper subsets and receptors
T lymphocytes originate from hematopoietic stem cells in bone marrow and migrate to the
thymus, a gland they derive their name from, to mature. There are two major subsets of T
cells (identified by their ‘cluster of differentiation’ or ‘CD’):
- CD4+ "helper" T cells, and
- CD8+ "killer" T cells.
T helper cells (Th)
T helper (Th) cells, which 'help' other immune cells in neutralizing infections. They recognize
a potential pathogen in the form of an antigen displayed in the MHC II complexes on the
surface of antigen presenting cell via their highly variable T cell receptors (TCR). T cells play
an essential role in the recognition of extracellular pathogenic microorganisms (certain
bacteria, protozoa, helminths).
White Blood Cells
- Function: T cells recognize antigens (substances that prompt an immune response) that
the body has encountered before. However, unlike another immune cell called a B cell, T
cells do not make antibodies, or proteins that correspond to specific antigens and help
fight infection. Instead, some T cells help B cells make antibodies and others directly kill
an infected cell or a cancerous cell. T cells also help prevent the immune response from
attacking normal host cells inappropriately.
- Location: T cells originate in the bone marrow and become specialized in the thymus, a
small organ just under the breastplate. They then recirculate between the bloodstream
and the lymphatic system and patrol the tissues. T cells are also called T lymphocytes.
T helper cells are a subset of T cells that express the CD4 co-receptor. They are an essential
component in the development of an adaptive immune response. The two main populations
of T lymphocytes are CD4 T helper cells and CD8 cytotoxic or killer T cells.

Both cell types have antigen specific receptors called T cell receptors on their cell surface,
which recognize small protein fragments called peptides bound to major histocompatibility
complex proteins or MHC molecules. MHC Class II molecules are generally only expressed by
professional antigen presenting cells, such as dendritic cells. The peptide fragments presented
by MHC Class II molecules are derived from exogenous or extracellular proteins. For example,
a dendritic cell which phagocytosis and engulfs bacterial antigens will degrade these bacterial
proteins into short peptide fragments, which can then be presented to CD4 helper cells on
MHC Class II molecules.

When a naive CD4 T cell is activated by an antigen presenting cell, there are multiple outcomes
that are possible. Examples of T helper subsets include follicular helper T cells as well as Th1,
Th2, Th17, Th22, Th9 and inducible regulatory T cells. Following activation, the type of T
helper cell subset that develops plays a key role in defining the subsequent adaptive immune
response.

The term T helper cell came from the observation that generation of an efficient B cell
antibody response requires T cell help. This B cell help is provided by a specialized subset of
CD4 T cells named follicular helper T cells. Once they have encountered an antigen, dendritic
cells migrate to T cell zones of secondary lymphoid organs such as lymph nodes, where they
activate naive CD4 T cells. Production of the cytokines IL6 by the antigen presenting cell drives
expression of the transcription factor BC IL6 in the activating T helper cell and results in
commitment to the follicular helper T cell lineage.
Follicular helper cells then express high levels of the chemokine receptor, CXCR5, which
enables them to migrate towards the B cell zone of the lymphoid organ. Here, follicular helper
T cells can provide a costimulatory signal to the differentiating B cells and secrete the cytokine
IL21, which further drives B cell proliferation.

In addition to their well characterized role in supporting antibody responses, T helper cells
are also known to be important in the generation of effective CD8 T cell memory. For example,
studies of viral infection in mice that cannot mount a T helper response because they don't
have any MHC Class II molecules, find that the numbers of virus specific CD8 T cells are
dramatically reduced. T helper cells support a CD8 T cell response by conditioning antigen
presenting cells to deliver costimulatory and cytokine signals to the cytotoxic T cell. However,
there is also evidence that in some cases, direct interactions between T helper and CD8 T cells
may also be important. T cell development occurs in the thymus, but CD4 cells don't emerge
from the thymus as one subtype or another, but as a naive CD4 T cell whose fate is not yet
determined, and which can be heavily influenced by the microenvironment. Cytokines
present in the local environment can polarize differentiating CD4 T cells, pushing them to
become one sub-type over another.

For example, presence of the cytokine IL12 results in polarization into Th1 cells that express
the Th1 defining transcription factor T-bet on which secrete interferon Gamma. The presence
of IL4 on the other hand results in polarization into Th2 cells that express the transcription
factor GATA3 and which secrete Th2 cytokines such as IL4, IL5, and IL13. Whilst the local
cytokine environment is one of the key drivers of T helper cell polarization, which is by no
means the only factor. Many different elements act synergistically as well as antagonistically
to achieve polarization of a T helper immune response, so that it is hopefully appropriate and
useful.

Th1 immune responses and production of the pro-inflammatory cytokine interferon Gamma
are typically associated with responses to bacteria and viruses. But there are also situations
in which a polarized T helper response is damaging to the host and can be a cause rather than
a resolution of disease. In the pathogenesis of type 1 diabetes for example, a model of Th1
driven pathology has been proposed where polarized Th1 cells drive the destruction of
pancreatic Beta cells through recruitment and activation of other inflammatory cells. There's
much data to support this model, but also other studies that suggest that this model is too
simplistic and the other T helper subsets also play a role.

Another example of a disease that was historically described as Th1 mediated is multiple
sclerosis. However, in the late 1990s, studies using a mouse model demonstrated that mice
lacking interferon Gamma or IL-2 signaling could still be susceptible to MS like disease. In 2005
a new T helper subset termed Th17 was described. Th17 cells were shown to be able to drive
auto-immune inflammation. And their discovery was a huge leap forward in our
understanding of the complexity of T helper cell biology. We now know that naive CD4 T cells
can be polarized towards a Th17 phenotype by the presence of cytokines such as TGF Beta
and IL-6 and that they produce the effect of cytokine IL-17. The transcription factor ROR
Gamma T is the master transcriptional regulator of this cell phenotype and Th17 cells have
been implicated in many studies as playing a key role in both health and disease.

Th2 responses are commonly generated in response to helminths and other extracellular
parasites, but inappropriate development of a Th2 immune response can also lead to
pathology. Allergic asthma is the prototypic example of a Th2 mediated disease. Many studies
show activated Th2 cells to be highly abundant in the airways of patients with allergic asthma.
The Th2 effector cytokines, IL-4, IL-5, and IL-13 can both initiate and maintain many of the key
pathological features of this disease. Other more recent T helper subsets that have been
described include Th22 and Th9 cells. Th9 cells can produce several prototypic Th2 cytokines,
such as IL-5 and IL-13, but also secrete high levels of IL-9. They have been linked to defense
against parasites as well as in the pathogenesis of allergic asthma and some auto -immune
diseases. Th22 cells are more closely related to the Th17 subset. Like Th17 cells, they are
polarized by IL-6 and secrete IL-22, but they do not secrete IL-17. Th22 cells are associated
with antimicrobial responses and have been demonstrated to have protective effects in
several disease models, such as asthma and inflammatory bowel disease.

Much of T helper cell biology depends upon the interpretation of environmental cues to direct
context appropriate T helper cell subset development. One of the more important examples
of this challenge relates to the balance between Th17 cells and peripheral or inducible T-REx.
Development of both is driven by the presence of TGF Beta in the local cytokine micro -
environment. However, the downstream actions of these two cells are for the most part the
opposite of each other, with Th17 cells being pro-inflammatory and associated with
autoimmunity while peripheral T-Rex are anti-inflammatory and play an important role in
maintaining immune tolerance. It is the additional presence of pro-inflammatory cytokines,
IL-6 that tips the balance in favor of Th17 differentiation. At the start of our understanding of
T helper cell subsets, it was believed that subset differentiation was a permanent state. But
there are now many examples of T helper cells being more flexible in their phenotypes, and
they are producing different combinations of cytokines at any one time, such as simultaneous
secretion of IL-17 and interferon Gamma. Understanding the rules governing plasticity versus
stability of T helper subsets is key to a more complete understanding of the role that these
cells play in adaptive immunity.
T helper cell subsets and their functions
Here are the T helper cell subsets, their associated transcription factors, and their functions:
- TfH (Bcl6) helps B cells in antibody production, affinity maturation and antibody class
switching.
- Th1 (T-bet) fights against intracellular invaders, activates the action of cytotoxic T cells and
microphages.
- Th2 (Gata3) fights against extracellular invaders, stimulates naïve B cells to attack invaders
via antibodies.
- iTreg (Foxp3 supports immune homeostasis, and suppressive functions in immune
tolerance.
- Th9 promotes T cell proliferation, along with IgE and IgG production.
- Th17 (RORyδ) defend against extracellular bacteria and fungi.
- Th22 protects epithelial barrier organs (e.g. skin, lungs), and modulates inflamed and