Immunology Exam 1 Flashcards

(113 cards)

1
Q

Contrast innate and acquired immunity (4).

A
  • nonspecific VS specific
  • common to all living things VS vertebrates
  • present at birth VS developed over time
  • no memory VS memory
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2
Q

Examples of innate system elements (5).

A
  • Physiological barriers
  • PMLs
  • Macrophages
  • NK cells
  • Inflammation
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3
Q

endocytosis

A

ingestion of foreign macromolecules by pinocytosis or receptor-mediated endocytosis.

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4
Q

phagocytosis

A

ingestion and destruction of foreign particles.

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5
Q

opsonin

A

facilitator of phagocytosis. May be an antibody (ex IgG) or complement.

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6
Q

reticuloendothelial system

A

includes macrophages, dendritic cells (spleen and lymph nodes), interdigitating cells of thymus, Langerhans cells in skin, and other APCs throughout the body.

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7
Q

Describe the relationship between phagocytes and the acquired system.

A

Phagocytic cells take in antigens, process them, and present them to the lympocytes of the acquired immune system so that a specific acquired immune response (cellular and humoral) can be initiated against that antigen.

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8
Q

(1) Describe the inflammatory response including (2) the hallmarks of inflammation and (3) which cells are involved.

A

Goal is to bring tissue back to normal. Triggered by exogenous or endogenous tissue damage.

Hallmarks: redness (rubor), pain (dolor), heat (calor), swelling (tumor) and loss of function

Cells: PMNs, macrophages and lymphocytes. PMNs show up in 30-60 mins. Mononuclears show up in 5-6 hours.

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9
Q

List the 3 main cell types involved in acquired immunity and briefly describe the function of each.

A

-T-cells: cell-mediated response. Direct attack on infected cells, tumor cells.
- B-cells: antibody production.
- Phagocytes: antigen processing and presentation.

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10
Q

Name the primary lymphatic organs and the type of cell that matures in each.

A

Thymus: maturation of T-cells
Bone marrow: origin of lymphocytes and maturation of B-cells.

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11
Q

Name the 2 major secondary lymphoid organs and 2 major functions that occur in these organs.

A
  • Spleen
  • Lymph nodes

(1) trap and concentrate foreign antigens, and (2) main site of production of antibodies and antigen-specific T-cells

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12
Q

Describe the circulation pathway of lymphocytes.

A

Circulate through blood, and enter lymphatic system. Re-enter circulation through thoracic duct, which empties into right atrium. Use HEV to “home in”, enter appropriate tissues through endothelial cell barrier.

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13
Q

Antigen

A

any agent capable of binding specifically to components of immune response.

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14
Q

Immunogen

A

any agent capable of producing an immune response.

All immunogens are antigens, but some antigens can bind without causing an immune response.

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15
Q

Hapten

A

Non-immunogenic low MW molecules

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16
Q

Adjuvant

A

present in vaccines and enhance immunogenicity.

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17
Q

Epitope/Antigenic determinant

A

5-7 AAs long and serve as antibody binding sites. Do not “fit” into the conformation or chemical makeup of body’s constituents.

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18
Q

Valence

A

number of epitopes on a molecule

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19
Q

Cross reaction

2 definitions

A

An immunologic reaction in which cells or antibodies react with 2 molecules that have common epitopes but are otherwise dissimilar.

Cells or antibodies with specificity for 1 epitope react weakly with another epitope that structurally resembles it (but is not specific for it).

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20
Q

List the four requirements for immunogenicity.

A
  • Foreign to self
  • High MW
  • Chemical complexity
  • Degradability (in order survive and then to be presented)
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21
Q

List the 4 major classes of antigens and give an example of a diagnostically important antigen for each class.

A
  • Carboyhydrates—ABC antigens on erythrocytes
  • Lipids—cardiolipin
  • Nucleic acids—autoimmune response to DNA with SLE
  • Proteins—serum albumin
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22
Q

Contrast epitope recognition by B cells and T cells.

A

B cells use sIg (surface immunoglobulin) receptors to bind free, soluble antigen

T cells use TcR, which require processing and presentation in association with MHC

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23
Q

Contrast how foreign antigens entering the body through blood, skin and respiratory or gastrointestinal routes find their way to the lymphoid cells.

A

BLOOD: response initiated at spleen
SKIN: local inflammation; transport to lymph nodes by macrophages
GI: response initiated at MALT

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24
Q

examples of physiological barriers

A

skin and mucous membranes
fatty acids and low pH of sweat
sebaceous secretions
soluble proteins (interferons, complement)
respiratory tract (nasal hairs, cilia, cough reflex, mucus lining)
GI tract (salivary enzymes, low pH of stomach, enzymes in bile and small intestine)

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25
4 PMNs
neutrophils, eosinophils, basophils and mast cells
26
macrophages activated by...
cytokines released primarily from T-cells
27
differentiate NKs from lymphocytes
slightly more cytoplasm, granules
28
NK express... function
Express killer inhibitory receptors (**KIR**). KIR binds **MHCI** on normal cells and protects them from NK destruction. Tumor/virally infected cells have **abnormal MHCI** and cannot bind KIR. NK destroys them.
29
2 NK killing processes
(1) Creates pores in the membrane, causing lysis (2) Triggers apoptosis.
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trigger NK activity
IL-2 IL-12 interferons
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endogenous inflammation causes (2)
uncontrolled immune response abnormal clotting reactions.
32
inflammation releases vasoactive peptides, which cause... (4)
(1) smooth muscle **contraction**, (2) **nerve** stimulation, (3) **endothelial** cell contraction, and (4) expression of **ECAMs** (endothelial cell adhesion molecules) resulting in increased vascular permeability and recruitment of PMNs to the site.
33
liver proteins involved in inflammation. example.
acute-phase proteins **C-reactive protein** binds to membranes of certain microbes and activates complement.
34
Immunization
generation of subsets of lymphocytes that can recognize an antigen upon a secondary exposure
35
regions and cells of thymus
**Cortex** contains immature cells. **Medulla** contains more mature cells (which have their specific receptor)
36
atrophy of thymus
fatty metamorphosis
37
required for lymphocytes to enter appropriate tissues
homing in
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HEV
high endothelial venules
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typical MW of immunogens
>6000 Da
40
not good immunogens due to lack of variety
homopolymers
41
example of a hapten
cardiolipin
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homologous cross reactions
occur when antigen and immunogen are the same substance
43
heterologous cross reactions
occur when substance that induces immune response is something other than the antigen you are trying to elicit the response against
44
cross reactions cause...
false positives
45
# ★ Antibody
Product of the immune system that imparts protection. Belong to a protein class called **globulins**, collectively known as immunoglobulins. Exhibit a high degree of **specificity**, therefore one part of the molecule must be adaptable to a large variety of epitopes.
46
# ★ F(ab)
Product of **papain** degradation. Cuts above hinge. 2 F(ab) from each antibody.
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# ★ Fc
The "stem" of the "Y" below the hinge. Constant.
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# ★ Fd
Part of the heavy chains included on the Fab region.
49
# ★ F(ab')2
Product of **pepsin** degradation. Includes both Fab regions (cuts below hinge).
50
# ★ Allotype
different forms of the same protein. Changes in only 1 or 2 AAs. Genetic differences between different people.
51
# ★ Idiotype
different antigenic determinants expressed on variable regions.
52
# ★ Isotype
Type of heavy chains. Defines 5 classes of Ig.
53
# ★ Describe the structure of the immunoglobulin fragments resulting from cleavage with pepsin.
**F(ab’)2**. This contains both L chains and the Fd part of both H chains. It lacks the **Fc** of H chains, and therefore cannot do things like fix complement. The remainder of the molecule is broken down into smaller segments.
54
# ★ Describe the structure of the immunoglobulin fragments resulting from cleavage with papain.
**2 F(ab)**, containing one L chain and Fd part of one H chain, and **1 F(c),** containing remainder of the H chains. F(ab) has only one antibody combining site, and F(c) cannot react with antigen but retains biological properties of the Ig molecule.
55
# ★ Contrast the variable and constant regions of the heavy chains in terms of structure and function.
Differ in their antigenic activity, carb content, size and biological function. Variable regions define differences, constant regions offer the biological activity.
56
# ★ **IgG** - MW - half life - domains - cross placenta? - fix complement? - location - valence - subclasses - functions (5)
- 150,000 Da - 23 days - 4 domains - crosses placenta - fixes complement - predominant in blood, lymph, CSF, other fluids - valence 2 - 4 subclasses Functions: Can cause **precipitation** of multivalent soluble antigens. **Opsonin**. Activates **complement**. **Neutralizes** toxins and viruses. **Immobilizes** bacterial flagella and cilia by causing them to clump.
57
# ★ **IgM** - MW - half life - domains - cross placenta? - fix complement? - location - valence - subclasses - functions (4)
- 900,000 Da - 5 days - 5 domains - does not cross placenta - fixes complement - predominantly in intravascular space - valence 5 - no subclasses Functions: **First** Ig produced in an immune response, either to infection or immunization (acute phase antibody). Efficient **agglutinating** antibody (forms bridges between antigens that are too distant to be bridged by smaller IgG molecules). Natural **isohemagglutinins** (naturally occurring antibody against RBC antigens). Most efficient **complement** fixing antibody.
58
# ★ **IgA** - MW - half life - domains - location - valence - subclasses - functions (3)
- 160,000 Da monomer, 400,000 Da dimer - 4-7 days - 4 domains - does not cross placenta - secretory antibody - 4 valence - 2 subclasses Functions: Provides protection to **neonate** through breastmilk. can prevent attachment of organisms to cells and thus provide defense to **local** infections in the respiratory and GI tracts. Prevents viruses from entering host cells. Good **agglutinating** antibody.
59
# ★ **IgE** - MW - half life - domains - location - valence - subclasses - functions
- 200,000 Da - 2 days - 5 domains - low in serum, stays bound to B-cells - 2 valence - no subclasses Functions: “Reaginic”, protects against **parasitic** infections and binds to mast cells which causes hypersensitivity (**allergies**).
60
# ★ **IgD** - MW - half life - domains - location - valence - subclasses - functions
- 180,000 Da - 2.8 days - 4 domains - surface of maturing B-cells - valence 2 - subclasses unknown Functions: **Unknown** in serum. Does not appear to have a protective function. Does it have a function in B-cell maturation?
61
# ★ Compare and contrast IgG and IgM in terms of protection against infection and in which stage of infection they appear.
IgM made first. Isotype switching occurs later, and B-cells begin to make IgG. During second exposure, a small amount of IgM is made and a huge amount of IgG is made.
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# ★ Explain why IgM is the best agglutinating antibody.
Highest valence.
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# ★ Assess the usefulness of analyzing fetal or newborn serum for IgG and IgM antibodies.
Any fetal IgM indicautes intrauterine **infection**; does not cross placenta. 3-4 months gestation shows a rapid increase in mother's **IgG** concentration. 5 months gestation IgM and IgA fetal synthesis begins. 3-6 months after birth, maternal IgG is gone, and the baby must produce its own.
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percentages of light chain types
60% kappa 40% lambda
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domains
segments of antibody created by the interchain bonds
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hinge region found between...
CH1 and CH2
67
AAs present in hinge region
Cys and Pro
68
structure that allows crosslinking
hinge region
69
# ★ Serology
utilization of the in vitro reaction between antigen and serum antibodies. Usually looking for antibody in patient serum, not antigen.
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# ★ Monoclonal antibodies
highly specific reagents consisting of homogeneous populations of antibodies, all with precisely the same specificity toward an epitope.
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# ★ Association constant
characterizes binding of an antibody with an epitope, expressed by: K=[AbH]/[Ab][H] Where H = hapten or antigen.
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# ★ Affinity
initial force of attraction between Fab and a single epitope.
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# ★ Avidity
overall binding energy between antibody and multivalent antigen.
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# ★ Zeta potential
electrical charges on surfaces of particles
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# ★ prozone
antibody excess that can prevent agglutination or precipitation
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# ★ equivalence zone
proportion of antibody:antigen is optimal for precipitation
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# ★ postzone
more antigen than antibody
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# ★ Give 3 examples of primary antigen-antibody interactions.
Van der waals forces electrostatic forces hydrophobic forces
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# ★ Summarize how cross-linking formation occurs, including the antigen and antibody requirements necessary for it to occur.
interactions between **multivalent antigens** and antibodies that have at least **2 combining sites** per molecule (also: secondary interactions)
80
# ★ Give examples of some of the physical conditions that can influence antigen-antibody association and state how they would affect the interaction (ie: hinder or enhance).
- pH - incubation time - ionic strength - steric hindrance - **shielding** effects (Na and Cl) - **temperature** (IgM cold-reacting, IgG warm-reacting)
81
# ★ Give 3 examples of secondary antigen-antibody interactions.
agglutination precipitation complement activation
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# ★ Compare and contrast direct vs. passive agglutination.
**Direct agglutination**: cross-linking of insoluble particulate antigen with antibody, resulting in visible clumping. **Passive agglutination**: cross-linking of antibody with soluble antigen attached to an insoluble particle (latex, RBCs, etc).
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# ★ Compare and contrast agglutination, precipitation and flocculation.
- **Agglutination**: antigen is a particle or attached to a particle (insoluble). - **Precipitation**: antigen is soluble. - **Flocculation**: interaction of soluble antigen with antibody, which results in the formation of a fine precipitate
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# ★ Examples of flocculation assays
RPR and VDRL assays for syphilis
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precipitin reaction
When the lattice reaches a certain size, the Ab-Ag complex loses its solubility and precipitates out of solution
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# ★ 2 assays with precipitate in solution and difference between them
**Turbidimetry**: Detector is in **direct** line with incidence light. Measures reduction in light intensity due to reflection, absorption or scatter due to the precipitate. **Nephelometry**: Measures light scattered at a certain **angle** between detector and incident light.
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# ★ Explain double diffusion.
Antigen and antibody are placed in separate wells and each diffuses outward from the wells (toward each other) until they reach an equivalence point, at which they precipitate out of solution and form a **precipitin band.**
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3 patterns of double diffusion results
Identity Non-identity Partial identity
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# ★ Explain radial immunodiffusion.
Well contains antigen at different concentrations and antibodies are distributed uniformly in agar gel. **Precipitin ring** forms instead of precipitin band. The diameter is directly proportional to the amount of antigen in the sample. Uses known standards to create a **curve**.
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3 sources of error in immunodiffusion assays
(1) overfilling wells, (2) nonlevel incubation or inadequate incubation time, and (3) drying out or contamination of gels.
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4 sources of error in electrophoresis
(1) application of current in the wrong direction or wrong voltage, (2) incorrect pH, (3) incorrect time, and (4) incorrect concentration of antigen and antibody.
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# ★ explain radioimmunoassay (RIA)
type of direct-binding immunoassay which uses isotopically labeled molecules (typically anti-human-antibody).
93
# ★ explain solid-phase immunoassays.
protein antigens are adsorbed onto a **surface** (ie. polystyrene bead). Patient antibody reacts with the antigen. Anti-human-antibody (conjugate) labeled with an **enzyme** are added which then bind to the antibody bound to the solid phase.
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Utilizes fluorescent-labeled antibodies to detect antigen or antibody.
immunofluorescent assay (IFA)
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# ★ most common fluorochormes used in IFA
**FITC** (fluorescein isothiocyanate), which fluoresces apple green color **TRITC** (tetramethyl rhodamine isothiocyanate), red orange color
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# ★ Direct vs Indirect IFA
**Direct** IFA involves reacting patient tissue on a slide (ie. skin or kidney biopsy) with fluorochrome-labeled specific antibodies. **Indirect** IFA involves reacting the patient serum (antibody) with a tissue or cell culture substrate on a slide, and then adding a fluorochrome-labeled anti-human-antibody conjugate.
97
explain immunochromatographic assay
the analyte is applied at one end of the strip and **migrates** by capillary action to the other end. As it migrates, the patient antigen binds with its specific antibody that has a colored label attached. Further down the strip a **monoclonal** antibody captures this complex to immobilize it to show a colored line.
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immunochromatographic assay is a type of....
rapid immunoassay
99
# ★ Explain immunofixation electrophoresis.
serum is placed in a well of a gel and an electric current applied to separate the proteins, then the proteins are precipitated into the gel with specific anti-serum, and stained for visualization.
99
radial immunodiffusion detects...
antigen
99
# ★ explain western blots.
antigens or a mixture of antigens are separated by electrophoresis in a gel. Separated material is then “blotted” onto nitrocellulose sheets, and antigen binds tightly. Patient serum containing antibody is applied to the sheet and binds specifically to the antigen. The strip is washed with anti-human-antibody (binds to any antibody) and stained for visualization of bands, and band pattern is compared to a control.
99
Immunofixation electrophoresis is used to detect...
monoclonal antibodies
100
Visualize antibody structure
101
Visualize pepsin vs papain degradation.
102
Primary vs secondary exposure, IgM vs IgG
103
Visualize types of Ig differences
104
Precipitin curve
105
Immunofixation electrophoresis
106
Western blot
107
Indirect vs Sandwich ELISA
108
Direct IFA
109
Indirect IFA
110
Visualize immunochromatographic assay