WVSOM Antibody lectures 5&6 immuno
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| What is a selectin? | cell adhesion molecule that helps with cell to cell interaction
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| What are integrins? | cell adhesion molecules that help with cell to cell interaction, it has two subunits alpha and beta
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| How can we determine if a vaccine has worked? | measure the amount of antibody against the vaccine
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| So what are immunoglobulins? | they are part of normal plasma proteins produced by B cells, if you take the blood serum or plasma and put in an electrical field the proteins will separate according to their size and charge, most of antibodies will be down near the negative end
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| What is an immunogen? | any substance capable of inducing an immune response
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| What is an antigen? | any substance that can serve as the target of an immune response
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| When you immunize someone you give what? | an immunogen
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| Can immunogen and antigen be the same? | sometimes
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| What are the determinants of immunogenicity? | in general it has to have large size, chemical complexity, solubility and biodegradability, and if its foreign
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| What is a Hapten? | a substance, usually very small, that alone is not immunogenic, but after binding to a carrier protein or cell, becomes immunogenic.
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| After the hapten is bound what does the antibody bind to? | to the hapten alone, the carrier, or both
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| What are a couple examples of hapten? | substance in poison ivy combining with proteins on your skin, penicillin combining with your own proteins to give an adverse drug reaction
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| What does an antibody recognize and what are the terms were using? | antigen
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| What is an epitope? | the actual molecular substances that your immune system will recognize and react against, they are on the antigen
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| Can antigens have multiple epitopes, if so does it have a name? | yes they can have different epitopes or multiple copies of the same epitope, this would be called a multivalent antigen
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| What types of epitopes what are they? | a linear or discontinuous sequence of proteins that antibodies recognize and react against, the linear is in a line, discontinuous is more 3d
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| Which would the antibody still recognize if it has been denatured, linear or discontinuous epitope? | linear, discontinuous is more of a 3d structure making it harder for the antibody to bind if denatured
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| You have a Hypothetical virus has a lot of different glycoprotiens each could be an antigen, how many epitopes does the antigens have to respond to? | each antigen can have several epitopes in which the immune system may respond to.
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| Are all epitopes recognized by the immune system to purge the antigen? | Some epitopes may protect the body in helping the body to eliminate the organism, some may not mean anything at all
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| What are the biological roles for antibody? | neutralization, opsonization, complement activation
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| What is neutralization? | antibody can bind to the surface of an organism to prevent it from binding to your cells to prevent infection, can bind to toxin produced by a bacterium to neutralize that
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| What is opsonization? | antibody can coat the surface allowing for phagocytosis
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| What is complement activation? | classical pathway, B cell produces antibody, antibody binds to the surface of antigen, C1 protein comes in complement is activated on that surface
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| What does the humoral immune response lead to? | antibody production response
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| What does the cellular immune response lead to? | production of T-cells and macrophages things other than antibody
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| Does the humoral and cellular immune response talk to one another? | yes they interact
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| Are the antibodies produced in the primary response the same as the antibodies produced in the secondary response? | no they are a little different
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| Which is the larger response to an immunogen, primary or secondary? | secondary is larger and faster
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| What is the formula for the basic protein chain structure of immunoglobulin? | 2H + 2L = Ig, 2 identical protein heavy chains, 2 identical protein light chains
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| Are the chains of antibody arranged in a linear sequence? | no they are folded back and forth to form globular domains
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| What make up the different regions of an immunoglobulin? | antigen binding sites, variable region, light chain (212 residues), hinge region, and heavy chain (450 residues)
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| The light chains have what two regions? | variable region and constant region
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| In the heavy chains what regions do they have? | one variable region and at least 3 constant regions
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| What are the proteins in the antibody held together with? | disulfide bonds
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| What is the hinge region? | gives molecule some flexibility so the arms can move relative to each other
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| What is the antigen binding site and what is it made of? | have two identical antigen binding site formed from the variable regions of the light and heavy chain coming together this forms a 3d structure that binds to the antigen
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| They have identified a portion of the antibody called the Fab what is it composed of? | variable regions and constant regions number one and the constant of the light chain, its they Y of one arm its what binds to the antigen
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| Where does the binding site come from how is it formed? | formed from variable region of a light chain and a variable region of a heavy chain
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| Fc region of the antibody is what, what is its importance? | you antibody can be acting as a bridge between the organism and some cell of the immune system that comes in to kill it, opsonization, the cell will have a receptor that recognizes the Fc portion of the anitbody binds through a Fc receptor
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| In general how many different types of light chains are there? | 2 lambda or kappa
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| What are the five major antibody classes? | IgG, IgM, IgA, IgD, IgE
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| What are the different classes of IgG? | IgG1, IgG2, IgG3, IgG4
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| Which antibody class will be predominant in the blood? | IgG
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| IgG1 and IgG3 tend to be better at activating what than the others? | complement
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| What the two classes of IgA? | IgA1, IgA2
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| Where would you find most of the IgA antibody? | mucosal membranes and body secretions (breast milk, tears, saliva)
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| Over all which antibody does the body make most of? | IgA but it cant be sampled as readily as IgG
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| What does IgM form when secreted? | monomer, and pentamer 5 of the immunoglobulin molecules hooked together by a J chain
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| What does IgA form when secreted? | monomer, dimeric have two of them hooked together by a J chain
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| Most of the IgA that you find on the mucous membranes and secretions will be in what form? | dimer form
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| What is the secretory component and where is it found? | found on the IgA antibody, helps it survive on the membranes and is a transport mechanism that gets it from place to place
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| What joins the immunoglobulins together in a pentameric IgM? | J chain
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| Which is generally the largest of the immuoglobulins? | IgM most of it will be introvascular it doesn’t get out of the blood stream very much
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| Which immunoglobulin will be able to get out of the blood stream into the tissues? | IgG
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| What is an Isotype? | variants present in all members of the species, eg. IgG1, IgG2, etc
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| What is an Allotype? | variants that differ between members of the species
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| What is an idiotype? | variants due to the hypervariable regions of antibody and TCell Receptor, the body is able to make millions of these each different
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| What determines the different types of antibody molecules? | heavy chain
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| The blood sample you take from your patient should contain from most to least immunoglobulin? | IgG> IgA> IgM> small amounts of IgD> IgE generally the least
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| Where are IgD predominately found? | receptor on surface of B cells
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| Which immunoglobulin will be increased if you have an allergic disease, asthma, allergic rhinitis, or parasites? | IgE
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| Which immunoglobulin tends to stay around longer and why? | IgG, because the others tend to get eat up
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| What is Complement fixation? | the ability to activate compliment by classical pathway
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| In order to activate the complement by the classical pathway what needs to be in close approximation? | a lot of the Fc regions so you need several IgGs to activate compliment
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| Which immunoglobuline is the best at activating complement by the classical pathway? | IgM because of the pentimeric shape, IgG 1 and 3 are good as well but still take several IgGs to activate complement
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| Which immunoglobulin is able to cross the placenta? | Only IgG is able to bind to a certain receptor in utero and cross the placenta into the developing fetus circulation
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| Where are the actual binding sites of immunoglobulin and what are they made from? | 3 hyper variable regions that stick out from the binding site of where the light and heavy variable regions come together
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| Where do the hypervariable regions lie in the light chains? | discrete loops of the structure, available for contact with antigen
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| In general what type of bonding takes place in antibodies? | not covalent, usually hydrogen bonding, electrostatic attractions, Van der Waals forces, and hydrophobic interactions
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| What does antigen recognition induce in t cells? | induces expression of effector molecules by the T cell, which activates B cell
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| Different antibody classes are determined by what? | heavy chains
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| Which of the antibodies are transported across the mucosa via a secretory component? | IgA
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| A 5 day old baby born to a mom with gravis disease affect muscle control shows signs of muscle weakness, which immunoglobulin is most likely causing the muscle weakness? | IgG because this is the only one that crosses the placenta
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| What does antigen recognition induce in t cells? | induces expression of effector molecules by the T cell, which activates B cell
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| What is Auto reactive antibody? | as with any aspect of the immune response you can have an inappropriate activation of antibody production, you can have antibody that act against yourself, example lupus
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| What is immunofluorescence? | a technique used to detect certain antibodies in tissue
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| What is direct immunofluorescence? | fluoresceinated antibody allowed to bind directly to antigen in tissue section, it fluoresces when UV is added and reveals a pattern of expression of the antigen
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| What is indirect immunofluorescence? | you have an antigen in the tissue sample, put an antibody to bind to that antigen, then put a second antibody that has a fluoresceinated tag to bind to the first antibody
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| Can an antibody be an antigen if so how do you detect it? | yes you can detect with another antibody
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| What is the purpose of vaccinations? | to expose people with a lesser or nonactive strain so if they come in contact with the strain they can initiate a secondary immune response
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| What is the classical method of monoclonal antibody production? | put antigen in animal, harvest spleen of animal, harvest through PEG, cell fusion, test for antibody positive wells, clone antibody producers
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| What are monoclonal antibodies? | antibodies we use nowadays that are clones of an antibody that was isolated for a specific antigen
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| What type of antibody is Rituxan and its target? | Chimeric; CD20
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| What type of antibody is Herceptin and its target? | Humanized; HER2 receptors
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| What type of antibody is Remicade and its target? | chimeric; TNF
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| What type of antibody is Synagis and its target? | Humanized; RSV protein
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| What type of antibody is Xolair and its target? | Humanized; IgE
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| What type of antibody is Campath and its target? | Humanized; CD52 on leukemia cells
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| What type of antibody is Tysabriand its target? | Humanized; Alpha integrin
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| Why is it important to have a humanized antibody? | so the body will not reject or attack the antibody
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| How do you humanize an antibody? | add human proteins to the antibody before cloning so it will have human characteristics but retains its antigen binding specificity
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| What is a ELISA? | a popular laboratory method for detecting specific serum antibody, used to detect and semi-quantitate antigen or antibody
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| What does serology do? | measuring antibody amount and type present
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| What does ELISA stand for? | Enzyme-linked immunosorbent assay
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| What type of reagent are you going to use to measure a patients antigen in the serum? | antibody
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| What is a western blot and what is it used for? | combination of electrophoresis, protein transfer and antigen-antibody binding is used to detect immune response to pathogens
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| What is the confirmatory test for HIV antibodies? | a western blot
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| What is the most common immune deficiency for the antibodies? | IgA
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| What is immunofixation electrophoresis? | buffered agarose gel electrophoresis followed by overlay with specific antibody
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| What is Zone electrophoresis do? | find patterns of serum immunoglobulin abnormalities in various diseases
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| If you have IgG myeloma what would you see in a zone electrophoresis? | reduced albumen and a spike at position gamma
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| If you have a patient with Waldenstrom’s macroglobulinemia where is the spike in zone electrophoresis? | IgM
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| If you have poly clonal hypergammaglobulinemia what do you see in a zone electrophoresis? | a smear with elevated antibodies
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