Unit 14: Effector response of B lymphocytes: antibodies

ANTIBODY ISOTYPES
Antibodies are the effector molecules of B lymphocytes. They are secreted by them and, in greater
abundance, by plasma cells. Antibodies are also called immunoglobulins (Ig). There are five isotypes of
antibodies: IgG, IgA, IgM, IgD and IgE, exposed in order of decreasing abundance.
The isotype of antibody found mostly in the blood of an infected patient can guide you about the type of
infection you have and when it occurred. Recall that B lymphocytes secrete almost exclusively IgM in the
first phase of activation, while in a later phase -in the germinal center- other isotypes of Ig can be produced
and the generation of plasma cells and memory B lymphocytes:
- The presence of abundant IgM indicates the first phase of an infection.
- The presence of IgG, IgE, or IgA indicates the end of a current infection or an old infection. In the
second case, the presence of antibodies is due to long-lived plasma cells that were generated in the
second phase of activation to prevent a possible second infection.
- IgD is very low in the blood and its function is unknown.
When comparing the affinity and avidity characteristics of the different antibody isotypes we find the
following:
- IgM binds to the antigen most avidly, as it is a pentameric protein and therefore possesses ten
antigen binding sites (each Ig has two binding sites for the antigen). However, antigen binding is
low affinity.
- IgG, IgE and IgA can bind to the antigen with high affinity.
- IgA shows greater avidity than IgG and IgE because it is a dimer and therefore presents four antigen
binding sites.
These comparisons are useful between IgM and the other Ig, since at any given time the respective
responses occur against the same antigen. However, it is more difficult to compare IgG with IgE or IgA since
they are usually generated against different antigens, even against different types of pathogens.
The effector function of antibodies depends on the region involved:
- The Fab (antigen-binding) region of the antibodies serves to neutralize the antigens. For example,
in the case of toxins, when the antibody binds to the toxin, it can no longer cause damage because
it is blocked. All antibody isotypes have this function.
- The Fc region serves two purposes:
o Activation of the add-in system; IgM and IgG isotypes possess this capability.
o Binding to Fc receptors present in different cell types. IgG binds to the Fcγ receptor, IgE binds to
the Fcɛ receptor, and IgA binds to the Fcα receptor. The binding of the antigen-antibody with
the Fc receptor has different consequences depending on the type of cell.

ROLE OF IGM
B lymphocytes of types B 1, B 2ZM (marginal zone of the spleen) and B 2F (follicular) and plasma cells
derived from them secrete IgM after the first phase of activation. The production of IgM is induced by any
type of pathogen, regardless of whether it is extracellular or intracellular, phagocytable or non-
phagocytable.
The binding of IgM to the antigen is of low affinity, since the B lymphocytes have not reached the affinity
maturation process. At the same time, it is a highly avidity binding since IgM is pentameric and therefore
has ten antigen binding sites.
The functions of IgM are neutralization and activation of the complement:
- Neutralization is done by the Fab region of the IgM; by binding to the antigen, IgM prevents the
action of that antigen. In this way, toxins, degrading enzymes or adhesion molecules of different
pathogens can be neutralized. In the case of adhesion molecules or adhesins, IgM neutralizes the
pathogen and prevents it from binding to host cells and, therefore, its penetration into the cell
 interior. After neutralization, the antigen must be removed. The binding of IgM and antigen
becomes an immunocomplex. This immune complex then binds to the complement system.
Erythrocytes are able to bind to the set formed by the immune complex and the complement
system, thanks to complement receptors. Subsequently, these erythrocytes circulate through the
blood and reach the liver and spleen. In these organs, erythrocytes yield the immunocomplex-
complement set to macrophages of the spleen and liver, which eliminate them by phagocytosis.
- Complement activation is carried out by the Fc region of the IgM antibody. First, the IgM antibody
binds to the surface of the pathogen. Second, complement is associated with the antigen-antibody
complex and activated. Finally, complement collaborates in the elimination of the pathogen
through three processes: 1) the opsonization of the pathogen for its subsequent phagocytosis, 2)
the lysis of the pathogen through the creation of pores on its surface, and 3) inflammation.

IGG AGAINST EXTRACELLULAR PATHOGENS AND AGAINST INTRACELLULAR PATHOGENS

IgG is generated in the second phase of activation, which is unique to 2F B lymphocytes. It has a very high
affinity for the antigen, since it occurs after the maturation of affinity and the change of isotype. However,
binding is not highly avid, as it only possesses two binding sites for the antigen. IgG are the most abundant
immunoglobulins in serum as they are synthesized against multiple pathogens, which can be extracellular
phagocytable or intracellular.

Role of IgG against phagocytable extracellular

pathogens.
IgG antibodies participate (a) in the opsonization of
pathogens by bridging antigens (Ag) on the surface
of the pathogen and Fcγ receptors (RFcγ) on the
surface of phagocytes. Other receptors involved in
phagocytosis are Scavenger receptors (RSc), PAMP
recognition receptors (RRRP) and complement
receptors (CR). IgG also (b) activates the
complement system, which in turn opsonizes and
creates pores in the pathogen and induces
inflammation. IgG also neutralizes (c).

Role of IgG in antibody-dependent cellular

cytotoxicity (CCDA) against intracellular pathogens.
IgG antibodies participate (a) in cellular cytotoxicity
against viruses, bridging viral antigens (Ag) present on
the surface of infected cells and Fcγ receptors (RFcγ)
present on the surface of NK lymphocytes. RFcγ and
altered HLA receptors are NK activating receptors.
Healthy HLA receptors are inhibitory receptors. If the
activating signals (+) are greater than the inhibitory
ones (−), CCDA is induced. Ac IgG also directly
neutralize pathogens (b) by binding to their antigens.

The effector functions of IgG against both types of pathogens are neutralization and immunity in the
newborn.
- Neutralization occurs similarly as with IgM. They bind to antigens via the Fab region and form
immune complexes.
 - Immunity in the newborn is provided by maternal IgG, as it is the only immunoglobulin capable of
crossing the placenta. This is a passive, uninduced immunity in the newborn and can last up to the
first six or nine months of life. From birth, the newborn generates its own antibodies.
- In addition, there are other functions performed by IgG exclusively against phagocytable
extracellular pathogens. They are the opsonization and activation of the complement. In both
cases, phagocytosis is favored and, in the latter, the lysis of the pathogen.
- Also, exclusively against intracellular pathogens, IgG induces cytotoxicity of the infected cell in a
process called antibody-dependent cellular cytotoxicity. This process happens as follows:
o Cells infected by some viruses transform when the virus, leaving the cell, leaves viral proteins
on the surface of the host cell.
o These proteins act as antigens on the cell surface. The antigens are recognized by Fc receptors
present on natural killer (NK) lymphocytes.
o By recognizing the antigen, NK lymphocytes are activated and induce the death of the infected
cell, by apoptosis.

IGE VS. HELMINTHS

IgE is the least abundant antibody in the blood. It is produced by 2F B lymphocytes after the change of
isotype and the maturation of affinity, so it is of high affinity. It has only two antigen binding sites, which
promotes not very high avidity binding. IgE is typically generated against helminths.
The effector functions of IgE are neutralization and antibody-dependent cell-type cytotoxicity for
helminths.
- Neutralization occurs against the degrading enzymes released by helminths, among other
substances.
- Antibody-dependent cellular cytotoxicity occurs as follows:
o IgE binds to antigens on the helminth surface.
o Antigens are recognized by their Fc region by receptors present on mast cells, eosinophils and
basophils.
o Such cells are activated and release the contents of the granules to destroy the helminth.

Role of IgE against helminths.

IgE antibodies participate (a) in cytotoxicity against helminths,
bridging antigens (Ag) on the helminth surface and Fcɛ receptors
(RFcɛ) on mast cells and eosinophils. Binding induces the activation
of mast cells and eosinophils, which release the contents of their
granules in the process of antibody-dependent cellular cytotoxicity
(CCDA). Ac IgE also neutralize (b).

IgA in front of diners

IgA is the second immunoglobulin in abundance in the blood. It is found in all mucous secretions since its
main characteristic is its ability to cross the epithelial cells of the mucous membranes from inside the
organism towards the lumen of the cavities covered by the mucous membranes. It is generated by 2F B
lymphocytes after the change of isotype and the maturation of affinity, so they are of high affinity. In
addition, they have high avidity, since they form dimers and can use four antigen binding sites.
 Role of IgA in mucous membranes.
IgA antibodies participate in the neutralization of
commensal or pathogens, because they can cross the
epithelial layer of cells from the subepithelial zone, to reach
the lumen of the mucous membranes (digestive, respiratory
and genitourinary ).

IgA is generated against both commensal microorganisms and pathogens. Its effector function is
neutralization. By passing through the mucosa, IgA is directed towards microorganisms and, by binding to
them, prevents them from entering the body.

APPLICATIONS OF ANTIBODIES IN DIAGNOSIS AND THERAPY

There are many applications of antibodies that make them useful in the diagnosis of diseases:
- Detection of present or past infections.
- Diagnosis of chronic immune-based and autoimmune diseases.
- Evolution of chronic diseases.
- Diagnosis of tumors.
The use of antibodies for diagnostic purposes is carried out with different techniques. In almost all of them,
the antibody binds to the antigen and then the antigen-antibody complex is quantified. Examples of some
of these techniques are ELISA (enzyme-linked immunosorbent assay) or indirect immunofluorescence.
Immunoscintigraphy administers antibodies labeled with a radioactive isotope detectable from the outside
to the patient, so the presence of antigens can be tracked in vivo.
In addition, antibodies are increasingly used in the treatment of various diseases. Monoclonal antibodies
are often used for this. They are so called because the same antibody is obtained in great abundance
against an antigen. The generation process is laborious, but offers excellent results.
The first monoclonal antibodies began to be used in the 80s. There are, for example, inflammatory
cytokine-blocking monoclonal antibodies used against rheumatoid arthritis. This is a systemic chronic
autoimmune disease that causes various characteristic injuries to the joints. Blocking the inflammatory
cytokine TNF-α (tumor necrosis factor α) with antibodies slows the progress of the disease. These
antibodies are also used after transplants to prevent rejection and in the treatment of some malignant
tumors. Many of these antibodies continue to be investigated today.