Lecture 4: Specific Immunology 1. Antibodies are also called Immunoglobulins. a. Secreted by differentiated B cells called plasma cells. b.

Contain two identical heavy and two identical light chains. i. Each has a variable region (N terminal) and a constant region (C-terminal)

c. 2. Light chains a. The constant region contains either a kappa or a lambda, not mixed. i. This constant region distinguishes the two classes of light chain. ii. Kappa is predominant in humans. 1. It demonstrates allotypy, this simply means that there are different forms of the same protein. a. Kappa chain allotypes referred to as Km. 3. Heavy chains and Antibody Isotype a. The constant region determines the antibody isotype. i. This region can be , , , , or . ii. Give rise to isotypes IgA, IgB, IgG, IgD, and IgE; respectively. 1. Depending on the constant region the biological activity of the antibody will differ. b. The variable region determines the Antibody binding site. 1. Depending on the makeup of the variable region the antigen binding capacity of the antibody will differ. ii. The antibody binding site amino acid makeup determines the specificity of the antibody.

4. Hypervariable regions exist within the variable region of both heavy and light chains. a. These regions are called the paratope and interacts with an antigens epitope, also called the antigenic determinant. b. These regions are unique to an antigens complementary epitope, i.e. they match up to one another. i. Therefore they are termed complementary determining regions (CDRs). 5. The strength of interaction between one antigen binding site and a monovalent (single) antigen is the affinity. 6. The avidity is the overall binding energy of all the antigen-binding sites with antigen. 7. Summarized, antibodies are bifunctional molecules depending on the portion of the antibody that is active. a. The constant region, Fc, contains the molecules biological activity. b. The variable region, Fab, contains the antigen binding site. 8. To figure out that antibodies were bifunctional molecules a famous experiment was performed. In the experiment an antibody was cleaved by papain and by pepsin. The resulting fragments helped us figure out the structure of an antibody. a. Papain yields three molecules, two single copies of an antigen binding region F(ab) and one crystallized Fc that cannot bind antigen. b. Pepsin yields one molecule with two antigen binding sites, Fab2 and the Fc portion is proteolytically degraded.

c. 9. Immunoglobulin A (IgA) a. Exists as a monomer, dimer, and trimer. b. Associated with the J chain and secretory piece. i. Binding to the secretory piece transports the IgA to the gut lumen. ii. The secretory piece breaks off and goes into the lumen with IgA. It NEEDS the secretory piece to survive in the MALT tissue. c. Found mostly in the gut and secretions including breast milk!

i. By being included in breast milk a child can be endowed with immunities. d. Activates Alternative complement 10. Immunoglobulin D (IgD) a. Exists only in a membrane bound form. i. Expressed by nave B cells along with IgM. b. Functions are under investigation. 11. Immunoglobulin E (IgE) a. Exists only in the monomeric form. i. Involved in allergic reactions and with helminths. b. Barely detectable in serum because it is bound to FcR waiting for cross linkage. If cross-linked by an allergen it releases the contents of the cell. 12. Immunoglobulin G (IgG) a. Exists only in the monomeric form. i. Involved in opsonization by phagocytes, ADCC, complement, and neutralization of viruses and toxins. b. Makes up 75% of the circulating immunoglobulins. c. The only Immunoglobulin that cross the placenta! d. There are four subtypes i. IgG1 highest concentrations ii. IgG2 iii. IgG3 half life of only one week, rest have three weeks iv. IgG4 lowest concentration, does not activate complement e. IgG activates classical complement but not as strongly as IgM. 13. Immunoglobulin M (IgM) a. Exists only in the pentameric form i. As a result it is the most potent activator of complement! b. Only isotype expressed on immature B cells and is also expressed on nave mature B cells along with IgD. c. The first antibody to a site of infection. 14. Antibodies are activated when the antigen binding sites are crosslinked. 15. Cross reactivity is when an antibody generated for a specific epitope binds to another very similar epitope. 16. Monoclonal antibodies a. Antibody molecules arising from a single clone that all recognize the same epitope. 17. Polyclonal antibodies a. The total antibody collection from all different clones. 18. There is a full section here about hybridomas, ELISAs, and monoclonal antibodies as therapeutic agents. This did not ever come up on any tests and is very convoluted. I would tag this as more for interest than anything else. 19. In some cases antibodies can act as antigens. a. Hypervariable region as an antigen i. Remember this region is normally called the paratope. ii. When it is an antigen it is called an idiotope.

1. Antibodies against this are called anti-idiotypic. b. Constant regions (Fc) as an antigen i. Remember this region is normally called the constant region. ii. When this is an antigen it is called an idiotype. 1. Antibodies against this are called anti-isotypic. 20. So why should you know idiotopes and idiotypes? a. This idea plays a large role in producing treatments for diseases that produce excessive amounts of antibodies. b. In most models a human antibody is introduced into an animal. The animal then mounts an immunologic response and produces antibodies to the human antibody. We can use these to clear antibodies from a human system either by Fc or Fab. c. If your goal is to create an anti-idiotypic (Fab) antibody you would: i. Put an antibody into an animal. 1. The Fc portion would be from the animal. 2. The Fab portion would be human. ii. The animals immune system would make antibodies that target the Fab portion of the antibody. iii. We can get these antibodies and use them to clear that specific paratope (Fab) from a human system. d. If your goal is to create an anti-isotypic (Fc) antibody you would: i. Put an antibody into an animal. 1. The Fc portion would be human. 2. The Fab portion would be from the animal. ii. The animals immune system would make antibodies that target the Fc portion of the antibody. iii. We can get these antibodies and use them to clear an entire isotype from a human system! 21. Rh antigens a. Patients can be either Rh+ or Rh-. b. The Rh antigen is a protein expressed on the surface of RBCs. c. If a pregnant woman is Rh- and her child is Rh+ and there is a placental bleed the mothers immune system may be introduced to the Rh+ antigen and her system will produce antibodies against it. i. This is not normally a problem for the current child, instead the second child is attacked. d. If the woman has another child and it is Rh+ her immune system through IgG crossing the placenta will attack the developing child. i. This leads to erythroblastosis fetalis or hemolytic disease of the newborn. [Anemia and Jaundice] 22. The blood system a. There are four types of blood: A, B, AB, and O. b. The name denotes whether or not the blood has a certain type of antigen. i. A contains A antigen. ii. B contains B antigen.

iii. AB contains A and B antigen iv. O has no antigen at all. c. Conversely depending on the type of blood you have you will have an antibody to the other type. i. A has antibodies to B. ii. B has antibodies to A. iii. AB has no antibodies because it contains both. iv. O has antibodies to both A and B. d. Universal donor = Blood type O i. You can give this blood to any patient because no antigen is present. As a result no matter whom you give the blood to it will not cause an immune reaction. e. Universal recipient = Blood type AB i. You can give this patient any kind of blood because it does not have antibodies to either A or B. As a result any blood can be give. Immunoglobulin high points: IgG the most prevalent immunoglobulin, activates classical complement, only one that crosses the placenta. IgM the only pentameric immunoglobulin, the most potent activator of classical complement IgE involved in allergic reactions and helminthes IgD this wont be the answer on the test, we dont know much about it. IgA present in monomeric, dimeric, and trimeric forms; has the secretory piece and is found in the digestive system, only one present in breast milk, activates alternate complement.