COURSE: Medical Microbiology, MBIM 650/720 - Fall 2007

TOPIC: Cytokines and Immunoregulaion Lecture 14

FACULTY: Dr. Mayer

Office: Bldg. #2, Rm B18
Phone: 733-3281
Email: [email protected]

TEACHING OBJECTIVES:

SUPPLEMENTAL READING:

Male et. al.: Immunology (7th Ed.).Chpt. 1, ???

KEY WORDS:

Cytokines and Immunoregulation

I. Overview

Cytokines are a diverse group of non-antibody proteins that act as mediators between
cells. They were initially identified as products of immune cells that act as mediators and
regulators of immune processes but many cytokines are now known to be produced by
cells other than immune cells and they can have effects on non-immune cells as well.
Cytokines are currently being used clinically as biological response modifiers for the
treatment of various disorders. The term cytokine is a general term used to describe a
large group of proteins but there are other terms that are commonly used to describe
particular kinds of cytokines. These include: 1) monokines, cytokines produced by
mononuclear phagocytic cells; 2) lymphokines, cytokines produced by activated
lymphocytes, especially Th cells; and 3) interleukins, cytokines that act as mediators
between leukocytes.

Cytokines are not typically stored as preformed proteins. Rather their synthesis is
initiated by gene transcription and their mRNAs are short lived. They are produced as
needed in immune responses. Many individual cytokines are produced by many cell
types and act on many cell types (i.e., they are pleotroipic) and in many cases cytokines
have similar actions (i.e., they are redundant). Redundancy is due to the nature of the
cytokine receptors. Receptors for cytokines are heterodimers (sometimes heterotrimers)
that can be grouped into families in which one subunit is common to all members of a
given family. Some examples are shown in Figure 1. Since the subunit common to all
members of the family functions in binding cytokine and in signal transduction, a
receptor for one cytokine can often respond to another cytokine in the same family.
 Thus, an individual lacking IL-2,
for example, is not adversely
affected because other cytokines
(IL-15, IL-7, IL-9, etc.) assume
its function. Similarly, a
mutation in a cytokine receptor
subunit other than the one in
common often has little effect.
On the other hand, a mutation in
the common subunit has
profound effects. For example, a
mutation in the gene for the IL-
2R gamma subunit causes human
X-linked severe combined
immunodeficiency (XSCID) Figure 1. Receptors for various cytokines.
characterized by a complete or
nearly complete T and B cell
defects.

One cytokine often influences the synthesis of other cytokines. They can produce
cascades, or enhance or suppress production of other cytokines. In addition, they can
often influence the action of other cytokines. The effects can be: 1) antagonistic; 2)
additive; or 3) synergistic.

Cytokines bind to specific receptors on target cells with high affinity and the cells that
respond to a cytokine are either: 1) the same cell that secreted cytokine (autocrine); 2) a
nearby cell (paracrine) or 3) a distant cell reached through the circulation (endocrine).
Cellular responses to cytokines are generally slow (hours) because they require new
mRNA and protein synthesis.

II. Categories of Cytokines

Cytokines can be grouped into different categories based on their functions or their
source but it is important to remember that because they can be produced by many
different cells and act on many different cells, any attempt to categorize them will be
subject to limitations.

A. Mediators of natural immunity – Cytokines that play a major role in the innate
immune system include: TNF-α, IL-1, IL-10, IL-12, type I interferons (IFN-α and IFN-
β), IFN-γ, and chemokines.

1. TNF-α – Tumor necrosis factor alpha is produced by activated macrophages is

response to microbes, especially the lipopolysaccharide (LPS) of Gram negative
bacteria. It is an important mediator of acute inflammation. It mediates the
recruitment of neutrophils and macrophages to sites of infection by stimulating

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 endothelial cells to produce adhesion molecules and by producing chemokines
which are chemotactic cytokines. TNF- α also acts on the hypothalamus to
produce fever and it promotes the production of acute phase proteins.

2. IL-1 – Interleukin 1 is another inflammatory cytokine produced by activated

macrophages. Its effects are similar to that of TNF-α . and it also helps to
activate T cells.

3. IL-10 – Interleukin 10 is produced by activated macrophages and Th2 cells. It is

predominantly an inhibitory cytokine. It inhibits production of IFN-γ by Th1
cells, which shifts immune responses toward a Th2 type. It also inhibits
cytokine production by activated macrophages and the expression of class II
MHC and co-stimulatory molecules on macrophages, resulting in a dampening
of immune responses.

4. IL-12 – Interleukin 12 is produced by activated macrophages and dendritic cells.

It stimulates the production of IFN-γ and induces the differentiation of Th cells
to become Th1 cells. In addition, it enhances the cytolytic functions of Tc and
NK cells.

5. Type I interferons – Type I interferons (IFN-α and IFN-β) are produced by

many cell types and they function to inhibit viral replication in cells. They also
increase expression of class I MHC molecules on cells making them more
susceptible to killing by CTLs. Type I interferons also activate NK cells.

5. INF-γ – Interferon gamma

is an important cytokine
produced by primarily by
Th1 cells, although it can
also be produced by Tc
and NK cells to a lesser
extent. It has numerous
functions in both the
innate and adaptive
immune systems as
depicted in Figure 2.

6. Chemokines – chemokines
are chemotactic cytokines
produced by many kinds Figure 2. Functions of IFN-γ.
of leukocytes and other
cell types. They represent a large family of molecules that function to recruit
leukocytes to sites of infection and play a role in lymphocyte trafficking.

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B. Mediators of adaptive immunity – Cytokines that play a major role in the adaptive
immune system include: IL-2, IL-4, IL-5, TGF-β, IL-10 and IFN-γ.

1. IL-2 – Interleukin 2 is
produced by Th cells,
although it can also be
produced by Tc cells to a
lesser extent. It is the major
growth factor for T cells. It
also promotes the growth of
B cells and can activate NK
cells and monocytes as
depicted in Figure 3. IL-2
acts on T cells in an autocrine
fashion. Activation of T cells
results in expression of IL-2R
and the production of IL-2.
The IL-2 binds to the IL-R Figure 3. Functions of IL-2.
and promotes cell division.
When the T cells are no longer
being stimulated by antigen, the
IL-2R will eventually decay and
the proliferative phase ends Figure
4.

2. IL-4 – Interleukin 4 is produced

by macrophages and Th2 cells. It
stimulates the development of Th2
cells from naïve Th cells and it
promotes the growth of
differentiated Th2 cells resulting in
the production of an antibody
response. It also stimulates Ig
class switching to the IgE isotype.
Figure 4. Stimulation of T cell proliferation
by IL-2.
3. IL-5 – Interleukin 5 is produced by Th2 cells and it functions to promote the
growth and differentiation of B cells and eosinophiles. It also activates mature
eosinophiles.

4. TGF-β – Transforming growth factor beta is produced by T cells and many other
cell types. It is primarily an inhibitory cytokine. It inhibits the proliferation of T
cells and the activation of macrophages. It also acts on PMNs and endothelial
cells to block the effects of pro-inflammatory cytokines.

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C. Stimulators of hematopoesis – Some cytokines stimulate the differentiation of
hematopoetic cells. These include GM-CSF which promotes the differentiation of bone
marrow progenitors, M-CSF, which promotes growth and differentiation of progenitors
into monocytes and macrophages and G-CSF, which promotes production of PMNs.

III. Cytokine Networks

Although the focus has been on the production and action of cytokines on cells of the
immune system, it is important to remember that many of them have effects on other cells
and organ systems. A schematic diagram showing some of the interactions in the
cytokine network is presented in Figure 5.

Figure 5. The cytokine network

IV. Immunoregulation

The magnitude of an immune response is determined by the balance between antigen-

driven activation of lymphocytes and negative regulatory influences that prevent or
dampen the response. Regulatory mechanisms can act at the recognition, activation or
effector phases of an immune response. Examples of regulation that have already been
discussed include: 1) recognition of antigen in the absence of co-stimulation resulting in
anergy; 2) recognition of antigen with CTLA-4 engagement of B7 resulting in down
regulation of T cell activation; 3) cytokines with stimulatory or inhibitory activities on
immune cells; and 4) idiotype/anti-idiotype interactions leading to stimulation or
inhibition of immune responses. In addition to these there are other ways in which
immune responses can be regulated.

A. Regulation by antibody (Figure 6) – Soluble antibody can compete with antigen

receptors on B cells and block or prevent B cell activation. In addition antigen
antibody complexes can bind to Fc receptors on B cells, sending an inhibitory signal
to B cells.

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 Insert Fig 11.5 from Male et al.

B. Regulation by regulatory T cells (Tregs) – Regulatory T cells (Tregs) are a recently

described population of cells that can regulate immune responses. They do not
prevent initial T cell activation; rather, they inhibit a sustained response and prevent
chronic and potentially damaging responses. They do no have characteristics of Th1
or Th2 cells but they can suppress both Th1 and Th2 responses.

1. Naturally occurring Tregs – The thymus gives rise to a CD4+/CD25+/Foxp3+

cells that functions as a Treg. These Tregs suppress immune responses in a cell
contact dependent manner but the mechanism of suppression has not been
established.

2. Induced T regs – In the periphery some T cells are induced to become Tregs by
antigen and either IL-10 or TGF-β. Tregs induced by IL-10 are
CD4+/CD25+/Foxp3- and are referred to as Tr1 cells. These cells suppress
immune responses by secretion of IL10. Tregs induced by TGF-β are
CD4+/CD25+/Foxp3+ and are referred to as induced Tregs. These cells
suppress by secretion of TGF-β

3. CD8+ Tregs – Some CD8+ cells can also be induced by antigen and IL-10 to
become a Treg cell. These cells are CD8+/Foxp3+ and they suppress by a cell
contact dependent mechanism or by secretion of cytokines. These cells have
been demonstrated in vitro but it is not known whether they exist in vivo.