Antibody function and genes Flashcards
What is antibody valence?
Valence refers to the number of antigenic determinants an antibody molecule can bind.
What is an epitope/antigenetic determinant?
Not all of an antigen binds specifically to an antibody; the part that actually interacts is usually 10 to 20 amino acids long, and is called an epitope or antigenic determinant. Typical proteins have several epitopes which elicit and bind to different antibodies.
Please describe precipitation and agglutination.
The large immune complexes that are formed at or near equivalence tend to become insoluble and fall out of solution. When the antigen is a molecule, the phenomenon is called precipitation; when it’s a cell or cell-sized particle, it is called agglutination.
What is affinity?
Affinity: The strength with which an antibody molecule binds an epitope (antigenic determinant) is called its affinity.
Discuss why a line of precipitate may form in agar gel when antigen and antibody diffuse towards each other.
If you take a layer of agar gel, cut two holes in it, and put antibody in one and antigen in the other, they will begin diffusing radially out of their wells: In the area between the two wells, the situation is similar to a quantitative precipitin test; there is antigen excess near the antigen well, and antibody excess near the antibody well, so somewhere in between, eventually, equivalence must be reached. But: what happens at equivalence? The complex precipitates. We go back next day and look at our agar gel, and see… …a line of precipitate between the wells! This technique is called immunodiffusion.
Compare and contrast precipitation and agglutination in terms of the nature of the antigens involved, and sensitivity of the tests.
Agglutination is more readily detected than precipitation, and so an agglutination test is more sensitive. When the antigen is a molecule, the phenomenon is called precipitation; when it’s a cell or cell-sized particle, it is called agglutination.
List the components of complement in the order in which they become activated in the classical pathway. Name those that are also activated in the alternative pathway.
Complement is a large number of proteins, similar to the blood clotting system in that each exists in an inactive form, and when the first is activated the rest follow in a sort of cascade.There are at least three ways to activate the C cascade; the one that is most familiar is the classical pathway. More recently, an alternative and a lectin pathway have been described. Each pathway gets started differently â–ºbut all come together by C5.The classical pathway is activated by complexes of IgG or IgM antibody with antigen. There is a change in the Fc portions of the antibodies after interaction with antigen, which allows the binding and activation of C1q. C1 activates C4 and then C2, which together activate C3, which can then activate C5. â–ºClassical C counts: 1-4-2-3-5-6-7-8-9.
Discuss the different ways in which complement is activated by IgG and IgM. Identify the complement components which are: opsonizing; lytic; anaphylatoxic; and chemotactic.
â–ºThe C1q must interact with two Fcs simultaneously; it does so either by finding two IgGs close together, or a single IgM (this reinforces the point that IgM is a much more efficient a C activator than is IgG).It is LYTIC, as just described, if the membrane attack complex is activated. Neisseria (gonorrhea, meningitis) are by far the most susceptible family of bacteria to C lysis.It is OPSONIZING. One split product of activated C3, namely C3b, adheres to membranes. Phagocytic cells (PMN, macrophages) have C3b receptors, and so can get a firm grip on an antigen if it is opsonized with C3b. As we said before, IgG is also opsonizing, because phagocytes have receptors for its Fc end called FcR (there are several different FcR).It is CHEMOTACTIC. The C5 activation product, C5a, is chemotactic for phagocytes, especially neutrophils. This explains much of the inflammation in an antibody-mediated reaction, and why PMN are the hallmarks of such a reaction.It is ANAPHYLATOXIC. C3a, C4a and C5a are all capable of releasing histamine non-specifically from mast cells or basophils. This means that there will be increased blood flow in the area of antigen deposition, and a better chance for inflammatory cells to get out of the blood and into the tissues. Sometime, a person with a lot of complement activation will break out in hives, and you can confuse what’s going on with an allergic reaction.
Discuss how complement is important in immunity to bacteria even if the bacteria are resistant to lysis by C9. Identify the family of bacteria for which lysis is necessary for their clearance.
Remember - complement is both chemotactic and opsonizing. If it can’t lyse it’s subject it sure as hell can call in the guns (PMN’s and such).Regardless, here is how lysing works: The formation of the membrane attack (lytic) complex: C5, activated by any of the three pathways described, activates C6, C7, C8, and C9. C8 and C9 form a lesion on the target cell membrane which, on electron microscopy, looks like a hole; the cell loses its ability to regulate its osmotic pressure and lyses or pops.Nyseria: gonorrhea and meningitis
Discuss how complement plays roles in both innate and adaptive immunity.
The alternative pathway seems to be a more primitive, early, less-specific sort of defense, since it can work even without waiting for antibody to be made.There is also the lectin pathway of complement activation, truly part of innate immunity. The lectin pathway is mediated by mannose-binding protein, MBP or MBL, a lectin. Lectins are proteins that bind (usually foreign) carbohydrates. MBP binds certain mannose–containing structures found in carbohydrates of bacteria but not humans.Adaptive: B-cells, antibodies, and the activation of the classical pathway.
Describe how the lectin pathway of complement activation is triggered.
MBP is functionally similar to C1q in the classical complement pathway, so the lectin pathway goes MBP-4-2-3-5-6-7-8-9.
Please define toxoid.
A toxoid is a bacterial toxin whose toxicity has been weakened or suppressed either by chemical (formalin) or heat treatment, while other properties, typically immunogenicity, are maintained.
Please describe DNA recombination.
DNA recombination refers to the process that a DNA segment moves from one DNA molecule to another DNA molecule. Changing the relative positions of two pieces of DNA is called recombination.
Please define primary rna transcript.
A primary transcript is an RNA molecule that has not yet undergone any modification after its synthesis.
Please describe RNA splicing.
Splicing is a modification of an RNA after transcription, in which introns are removed and exons are joined.