Structure and some properties of ig classes and subclasses

IgG

Structure The structures of the IgG subclasses are presented in figure 7. All IgG's are monomers (7S immunoglobulin). The subclasses differ in the number of disulfide bonds and length of the hinge region.

Properties IgG is the most versatile immunoglobulin because it is capable of carrying out all of the functions of immunoglobulin molecules.

a) IgG is the major Ig in serum - 75% of serum Ig is IgG

b) IgG is the major Ig in extra vascular spaces

c) Placental transfer - IgG is the only class of Ig that crosses the placenta. Transfer is mediated by a receptor on placental cells for the Fc region of IgG. Not all subclasses cross equally well; IgG2 does not cross well.

d) Fixes complement - Not all subclasses fix equally well; IgG4 does not fix complement

e) Binding to cells - Macrophages, monocytes, PMNs and some lymphocytes have Fc receptors for the Fc region of IgG. Not all subclasses bind equally well; IgG2 and IgG4 do not bind to Fc receptors. A consequence of binding to the Fc receptors on PMNs, monocytes and macrophages is that the cell can now internalize the antigen better. The antibody has prepared the antigen for eating by the phagocytic cells. The term opsonin is used to describe substances that enhance phagocytosis. IgG is a good opsonin. Binding of IgG to Fc receptors on other types of cells results in the activation of other functions.

 IgM

Structure The structure of IgM is presented in figure 8. IgM normally exists as a pentamer (19S immunoglobulin) but it can also exist as a monomer. In the pentameric form all heavy chains are identical and all light chains are identical. Thus, the valence is theoretically 10. IgM has an extra domain on the mu chain (C_H4) and it has another protein covalently bound via a S-S bond called the J chain. This chain functions in polymerization of the molecule into a pentamer.

Properties

a) IgM is the third most common serum Ig.

b) IgM is the first Ig to be made by the fetus and the first Ig to be made by a virgin B cells when it is stimulated by antigen.

c) As a consequence of its pentameric structure, IgM is a good complement fixing Ig. Thus, IgM antibodies are very efficient in leading to the lysis of microorganisms.

d) As a consequence of its structure, IgM is also a good agglutinating Ig . Thus, IgM antibodies are very good in clumping microorganisms for eventual elimination from the body.

e) IgM binds to some cells via Fc receptors.

f) B cell surface Ig  Surface IgM exists as a monomer and lacks J chain but it has an extra 20 amino acids at the C-terminus to anchor it into the membrane (figure 9). Cell surface IgM functions as a receptor for antigen on B cells. Surface IgM is noncovalently associated with two additional proteins in the membrane of the B cell called Ig-alpha and Ig-beta as indicated in figure 10. These additional proteins act as signal transducing molecules since the cytoplasmic tail of the Ig molecule itself is too short to transduce a signal. Contact between surface immunoglobulin and an antigen is required before a signal can be transduced by the Ig-alpha and Ig-beta chains. In the case of T-independent antigens, contact between the antigen and surface immunoglobulin is sufficient to activate B cells to differentiate into antibody secreting plasma cells. However, for T-dependent antigens, a second signal provided by helper T cells is required before B cells are activated.

 IgA

Structure Serum IgA is a monomer but IgA found in secretions is a dimer as presented in Figure 11. When IgA exits as a dimer, a J chain is associated with it.

When IgA is found in secretions is also has another protein associated with it called the secretory piece or T piece; sIgA is sometimes referred to as 11S immunoglobulin. Unlike the remainder of the IgA which is made in the plasma cell, the secretory piece is made in epithelial cells and is added to the IgA as it passes into the secretions (Figure 12). The secretory piece helps IgA to be transported across mucosa and also protects it from degradation in the secretions.

Properties

a) IgA is the 2nd most common serum Ig.

b) IgA is the major class of Ig in secretions - tears, saliva, colostrum, mucus. Since it is found in secretions secretory IgA is important in local (mucosal) immunity.

c) Normally IgA does not fix complement, unless aggregated.

d) IgA can binding to some cells - PMN's and some lymphocytes.

IgD

Structure The structure of IgD is presented in the Figure 13. IgD exists only as a monomer.

Properties

a) IgD is found in low levels in serum; its role in serum uncertain.

b) IgD is primarily found on B cell surfaces where it functions as a receptor for antigen. IgD on the surface of B cells has extra amino acids at C-terminal end for anchoring to the membrane. It also associates with the Ig-alpha and Ig-beta chains.

c) IgD does not bind complement.

 IgE

Structure The structure of IgE is presented in Figure 14. IgE exists as a monomer and has an extra domain in the constant region.

Properties

a) IgE is the least common serum Ig since it binds very tightly to Fc receptors on basophils and mast cells even before interacting with antigen.

b) Involved in allergic reactions - As a consequence of its binding to basophils an mast cells, IgE is involved in allergic reactions. Binding of the allergen to the IgE on the cells results in the release of various pharmacological mediators that result in allergic symptoms.

c) IgE also plays a role in parasitic helminth diseases. Since serum IgE levels rise in parasitic diseases, measuring IgE levels is helpful in diagnosing parasitic infections. Eosinophils have Fc receptors for IgE and binding of eosinophils to IgE-coated helminths results in killing of the parasite.

d) IgE does not fix complement.

9. Describe general characteristics of the specific immune response;

10. Methods of antigen administration to mice.

Manual restraint of a mouse using both hands.

The mouse is lifted by the base of the tail and placed on the cage lid or other solid surface with one hand and then its tail is pulled gently back. It is quickly and firmly picked up by the scruff of the neck behind the ears with the thumb and index finger of the other hand. The tail is transferred from the first hand to between the palm and little or ring finger of the other hand, then fixed. The mouse is restrained.

Preparation of the site

The area for administration is clipped or cleaned with warm water if necessary before cleaning the skin with alcohol- or disinfectant-moistened cotton. Where aseptic skin is necessary; the fur must be clipped followed by a three-stage surgical preparation: surgical soap, alcoholic rinse and surgical preparation solution. The skin is dried immediately before administration

Method intravenous (IV) administration of antigen to mice

Compounds that are poorly absorbed by the digestive tract may be given intravenously. The syringe plus needle or the catheter must first be filled with the solution to remove air bubbles. Administrations are usually made into the lateral tail veins not into the dorsal tail vein, as it is not straight. The lateral veins are readily visualized, but have quite small diameters.

The mouse is either placed in the restrainer or anesthetized and the tail is then armed with a lamp or warm towel, or immersed in warm water (40–45ºC) in order to dilate the vessels. The tail is swabbed with 70% alcohol on a gauze sponge or swab. Insert the needle parallel to the tail vein penetrating 2–4 mm into the lumen while keeping the bevel of the needle face upwards. The solution is then injected slowly and no resistance should be felt if the solution is properly administered. The injected solution temporarily replaces the blood but then should be washed away by the blood stream. If this does not happen the position of the needle is certainly not in the vein but in the surrounding tissue so it must be moved in the surrounding tissue in such a way that it then enters the vein or a new try must be made. When the intravenous administration is finished or the cannula is pulled out, the injection site must be pressed firmly with a swab or fingers to prevent backflow of the administered solution and/or blood . The recommended volume is ˂0.2 ml.

The ophthalmic plexus route is also used for intravenous administration. The technique resembles the blood collection by retroorbital sinus puncture .