antigen structure and immunogenicity updated: september 22, 2015 folder title: antigens (with tp...
TRANSCRIPT
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Antigen Structure and Immunogenicity
Updated: September 22, 2015
Folder Title: Antigens(With TP Slides)
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Antigen Recognition by the Specific Immune System:Recognition by Antibodies on Bone-marrow Derived Lymphocytes (B-Cells)
or by Antibodies Secreted by B-Cells (Plasma Cells)or
Recognition by T-Cell Receptors on T-Cells (Thymus-derived Lymphocytes)
Antigen (Ag): Binds specifically to an antibody binding site (Ab), or to a T-cell receptor (TCR)*
• (* When the antigen is being presented to the TCR on a specific set of cell-membrane proteins called Major Histocompatibility Proteins (MHC)
Immunogen: Binds specifically to an antibody binding site or to a T-cell receptor*,
and generates a humoral or cellular immune response.
All immunogens must be antigens, Not all antigens can generate a response.
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Haptens and Epitopes Haptens: Small organic molecule
(e.g. 1-Fluoro-2,4-dinitrobenzene)
Not Immunogenic by themselves
Can be immunogenic when attached to an immunogenic macromolecular carrier
The Anti-hapten antibody so generated will bind the Hapten
Epitope: A region of a macromolecular antigen that is actually an antigenic determinant,
a molecular sub-region that is actually recognized and bound by an Antibody or T-Cell Receptor
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Fig.4-6bKuby3rd Ed
AgAb
Sep
Influenza Virus AntigenAntibody Heavy ChainVariable Region
Antibody Light Chain Variable Region
Antigen-Antibody Complementarity
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Antigen - Antibody BindingAntibody Light Chain
Variable Region
Fig.
4-6a
Kuby
3rd Ed
AgAb
KissInfluenza Virus Antigen
Antibody Heavy Chain
Variable Region
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Prot2nd
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Primary sequential epitopes in
sperm whale myoglobin
(Epitopes are antigenic determinants inMacromolecules)
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Conformational (non-sequential) Epitope in Hen
Egg White Lysozyme.
(Colors show amino acid side chains
contacting the two different chains of the
antibody, or contacting both
chains. Antibody structure to be covered later)
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Myoglobin Single Polypeptide
17,000 MW (Tertiary Structure)
About 7 secondary domains. One bound Heme in hydrophobic
Hemoglobin: a2b2 Tetramer64,000 MWOne heme per subunit
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“Fill-in-the Blank” Question:When a small organic molecule is made immunogenic
by attaching it to a macromolecule it is called a __ __ __ __ __ __
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Rank Responses
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6
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What is this molecular
model showing?
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What Determines Immunogenicity of an Antigen?
1.Properties of the immunogen(See detailed breakdown next slide)
2.Properties of the Host (Slides 12 and 18)
a. Geneticb. Physiological
3. Mode of Antigen Exposure
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ImGenFx1
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ImGenFx2
Called “H2” in Mice; “HLA” in HumanHLA = Human Leucocyte-associated Antigens
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Immunogenicity of Antigens and the MHC Gene Collection:MHC = Major Histocompatibility ComplexHaplotype = a set of closely linked genes controlling the immune response. Haplotype is usually transferred to progeny as a linked set
Antigen #1 Antigen #2
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Ag&MHCbAg 1 Ag 2
Two different antigens
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Ag&MHCcAg 1 Ag 2
Two different antigens
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Ag&MHCdAg 1 Ag 2
Two different antigens
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Ag&MHCAg 1 Ag 2
Two different antigens
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ImGenFx3
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ImAdmin
To here: Tuesday,Sept. 15, 2015
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Conformational Properties of Epitopes for Antibodies
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64
80
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Hen Egg-White Lysozyme: Conformations of Sequence 62 to 80
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Anti-hapten Antibodies
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AbHaptenSee Figure 4-1p. 77Kuby, 6th Edition
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AgByB&Ta
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AgByB&Tb
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AgByB&T
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Conformational (non-sequential) Epitope in
Hen Egg White Lysozyme.
To here: Thursday,Sept. 17, 2015
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0% 0% 0%0%0%0%
Response Counter
This graph shows two forms of the amino acid sequence between cysteine 64 and cysteine 80 in hen egg white lysozyme
(HEL). One form of the synthetic antigenic region does not inhibit the
normal reaction between HEL and its antibody while the other form does inhibit. Why does the form of the
potential antigenic sequence shown in green fail to inhibit the antigen-antibody reaction between HEL and its antibody?
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What does this table tell you about the specificity of antibody recognition for antigens?
Rank Responses
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To here: Thursday,Sept. 17, 2015
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Antigens in Immunologically-Based Clinical Diagnosis and Pathology
Cancer Antigens in Diagnosis of Cancer
Carcino-Embryonic Antigen (CEA)
Prostate Specific Antigen (PSA)
Ovarian Cancer Antigen (CA 125)
Whole-body Imaging of Metastatic Colon Carcinoma
A33 Antigen
Anti-Human Chorionic Gonadotropin Pregnancy Test(Early Home Pregnancy Test)
Type II Allergic Hypersensitivity
Blood Group Antigens in Transfusions
Hemolytic Disease of New-Borns: Rh Antigen incompatibility
Super-Antigens in Toxic Shock Syndrome
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Antigens in Immunologically-BasedTreatment of Cancers and Other Pathologies:
Cancer Antigens on Cancer Cells
Cancer Antigens as Targets for Therapy
Antibodies as Anti-cancer AgentsRadioactively-labeled Antibodies
Cytolytic Antibodies
Specifically-activated Cytotoxic T-Cells as Anti-Cancer Agents
Specifically Activated Antigen-presenting Cells (Tc).
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Imaging with Antibodies Carrying “Reporter Groups”
Radioactive IsotopesFluorescent Groups (“Chromophores”)Metals
Antigen-Antibody Interactions in Research and Clinical Medicine
e.g. Radioactively labeled monoclonal antibody reacting with tumor cell antigens in detection of metastatic colon cancer
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SeeMets
Arm
Head
Imaging on Metastatic Colon Carcinoma with Radioactive-Iodine-Labelled Monoclonal Ab to A33 AgLloyd Old, Scientific American, August, 1996, p. 138)
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Antihapten Antibodies in the Detection and Measurement of Small Molecules
For example: Anti-HCG (Human chorionic gonadotrophin) antibody in the detection of soluble HCG in urine
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AntiHCGWhat will happen when concentration of Anti-HCG-Ab is about equal to Hapten-carrier conjugate?
What will happen when you put excess soluble hapten in the way of the anti-HCG-Ab?
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PregTest
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Antibodies in Blood Transfusions
Antibodies to A, B, O Blood Group Antigens
Pre-existing Antibodies to Non-tolerated (Unmatched) Blood Group Antigens : “Isohemagglutinins”
Why are these non-matched Blood-group Antigens generating antibodies (Isohemagglutins) at all?
Because infectious bacteria do possess and exhibit cross-reacting antigens similar to blood group antigens
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ABO Types
Pre-existing serum antibodies from exposure to bacterial antigens that mimic antigens on red blood cells
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ABOSugar
See Figure 15-13 (a), p. 389,
Kuby, 6th EditionGalactose
Not a hexasaccharide(No terminal sugar)
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Antibodies in Hemolytic Disease of the New-born (HDNB)
(Rh-Factor Sensitization)
Rh-Factor positive baby – Rh Negative Mother
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HDNB
See Figure 15-14p. 390, Kuby6th Edition
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HDNBPhoto
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AntiRhAb
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Following are three Turning Point short answer questions.
Please put all notes on the floor.Do not have any electronic devices other than your NXT transmitter.No consulting with other students.
If you have a problem with your device, I can provide you with a loaner NXT device.
If you have a problem using your NXT device, please ask Elisabeth for help.
It is imperative that the integrity of these in-class Turning Point quizzes be maintained at the same level as we will do with the three written exams.
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“Fill-in-the Blank” Question:Hemolytic disease of the new-born is caused by maternal
__ __ __ __ __ __ __ __ __ __attacking the fetus in utero.
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“Fill-in-the Blank” Question:What are those maternal antibodies recognizing and reacting against?
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Rank Responses
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“Fill-in-the Blank” Question:What is this Whole-body X-ray Showing?
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Activation of Systemic Inflammatory Responses by Potent Inflammatory Signals
from Infectious Organisms and Plants
EndotoxinsExotoxinsSuper-antigensMitogens and Lectins
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Immunopathology of Bacterial Septic Shock Caused by Gram Negative Bacterial Endotoxins:Examples -E. coliPseudomonas aeruginosaNeisseria meningitidisMeningococcus
70,000 Deaths per yearDiarrhea, Fever, Blood Clotting, Blood Pressure Drop
Macrophage Activation by Endotoxin:Generates systemic cytokine production -IL1 and TNF-alpha (Tumor necrosis factor)
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Immunopathology of Bacterial Toxic Shock Exotoxins Secreted or Presented Membrane Bound:Act as "Superantigens"Examples -Staphylococcus aureus: TSST1 (Toxic Shock Syndrome Toxin)Streptococcus pyrogenes - Rheumatic fever and shockStaphylococcus enterotoxins: Food poisoning
Superantigens cross-link TCR with Antigen-presenting Cell MHC Class II Proteins
Generates systemic cytokine production - IL1 and TNF-alpha (Tumor necrosis factor)
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SuperAg
See Figure 11-67th Edition p. 367
Staphylococcal enterotoxins (food poisoning)Streptococcal pyrogenes exotoxins (rheumatic fever, toxic shock)See Table 11-2, p. 367, 7th Edition
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Mitogens and Lectins
Mitogens: Stimulate mitosis and cell division non-specifically(non-specific therefore stimulate polyclonally)
Lectins: Proteins that bind to carbohydrate in glycoproteins(Can be potent mitogens)
Glycoproteins: Conjugated protein with covalently attached carbohydrate residues
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Examples of Mitogens
Concanavalin A (Con A) : Non-specific T-cell mitogen
Phytohemagglutinin (PHA): Non-specific T-cell mitogen
Pokeweed Mitogen (PWM): Non-specific T- and B-cell Mitogen
Lipopolysaccharide (LPS): Gram negative bacterial cell wall component
Potent Polyclonal (non-specific) B-Cell Mitogen
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DescriptionGibco® Phytohemagglutinin, M form (PHA-M) is a solution made from a crude extract of the red kidney bean Phaseolus vulgaris. This extract is capable of inducing blastogenesis in vitro in various mammalian mononuclear cells when used at a concentration of 1–2% v/v. Each lot of Gibco® Phytohemagglutinin is performance tested at 1.5% using peripheral blood lymphocytes from a normal adult donor cultured for 72 hours in Gibco® RPMI 1640 Medium supplemented with Fetal Bovine Serum (FBS), heparin, antibiotics, and L-glutamine.
Google Description of Phytohemagglutin:What BIO 447 Is Intended to Achieve
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Immune Adjuvants and the Immunogenicity of Vaccines(See Scientific American, October, 2009)
Boosting Vaccine Power, pp 72 to 79
Activating Antigen-presentation by Cells of the Innate Natural Immune Response
e.g. Purified derivatives of bacterial lipo-polysaccharidesBacterial DNA derivativesBacterial flagellin protein
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Specific Activation of Antigen-presenting Cells Extra-corporeally (outside of the body)
Dendritic Cell VaccinesSee Dendreon
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Innate Natural Immunity and Systemic Sepsis:NOVA Video on Meningitis (Meningococcal
Meningitis, Endotoxin, and Systemic Sepsis)
http://www.pbs.org/wgbh/nova/meningitis/
Has video links to steps in meningococcal sepsis
“Amy’s Story”and related NOVA Stories
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tpfondy.mysite.syr.edu
The Story of the Oakwood Children
See “Song of the Oakwood Children” as a self-running presentation
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Following are two Turning Point short answer questions.
Please put all notes on the floor.Do not have any electronic devices other than your NXT transmitter.No consulting with other students.
If you have a problem with your device, I can provide you with a loaner NXT device.
If you have a problem using your NXT device, please ask Elisabeth for help.
It is imperative that the integrity of these in-class Turning Point quizzes be maintained at the same level as we will do with the three written exams.
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What is a “Mitogen”?What Does a Mitogen Do in Cell Biology Terms?
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“Fill-in-the Blank” Question:RhoGam is an antibody that prevents sensitization of the Rh- mother
by cells from her fetus. Why does that work?
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Merck Introduction of Keytruda
For Malignant Melanoma
Sept. 6, 2014Wall-Street Journal
Working Link Sent to Each Student by Yifan
Discussion and Comments
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Whooping Cough Infection and Transmission
Whooping Cough VaccineToday Show, 12/2/04
To Here September 16th; Showed 6 minute video of whooping cough disease and vaccinations
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Dendreon is a Commercial Vaccine that Uses Dendritic Cells to Generate an Immune Response. What is it that the Dendritic Cells do in Immunology that allow them to be used effectively in this way?
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End of Presentation on Antigens: Sept. 18, 2014
Save Session
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5. No problem. Please get moving!4. Maybe a bit too fast, but mostly OK.
3. I’m following OK, I can catch up with what I miss.4. I’m struggling to keep up. It’s too much too fast.
5. I’m lost(This slide is anonymous. I won’t know who you are)
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We watched parts of a video on Meningitis and Meningococcal Sepsis caused by an Infectious Bacterium. Killing the bacterium doesn’t immediately cure the sepsis. Why is that? What is causing the systemic sepsis?
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“Fill-in-the Blank” Question:The antibody to Human Chorionic Gonadotrophin fails to cross link the
HCG-hapten carrier conjugate and fails to make a precipitate if urine from a pregnant woman is added to the mixture. What is in the urine that
is preventing the formation of the precipitate? Abbreviations are OK______________________________________
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“Fill-in-the Blank” Question:Human chorionic gonadotrophin (HCG) is a small peptide made
immunogenic by being conjugated with a large molecular weight carrier molecule. Thus HCG is a special kind of antigen and is an example of a
__ ___ ___ ___ ___ ___
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“Fill-in-the Blank” Question:A small organic molecule such as fluoro-dinitro benzene
can be made immunogenic by attaching it to a _____________________________
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Immunogenicity of Antigens is Controlled by the Nature of the Antigen, the Genetics of the Host, and the Mode of Immunogen Exposure.
Give an example of what is meant by “Mode of Immunogen or Antigen Exposure”
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A Specific target for an Antibody is an_ _ _ _ _ _ _.
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