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Practice Flashcards

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1
Q

PAS reaction

A
  • Lymphoid cells - homogenous cytoplasm
  • Myeloid cells - granulous color
  • Can in some cases diagnose acute lymphoblastic leukemia (ALL) and also some subtypes of acute myeloid leukemia (AML)
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2
Q

Esterase reaction

A
  • Distinguish between myeloid cells, monocytes and neutrophil granulocytes
  • Used to identify monoblastic leukemias
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3
Q

Flow cytometry

A

Can characterize the differentiation and activation states of cell types by the determination of their surface and cytoplasmatic protein expression

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4
Q

Immunohistochemistry

A

Determination of antigens in tissue sections by the use of labeled antibodies

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5
Q

Indirect ELISA

A
  1. Add antigen to the plate to coat the wells
  2. Wash
  3. Add the biological sample (eg.blood serum)
  4. Wash
  5. Add labeled secondary antibodies
  6. Wash then add chromogenic substance for coloring
  7. Measure optical density
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6
Q

Sandwich ELISA

A
  1. Coat well with monoclonal Ab
  2. Blocking and washing
  3. Add antigen containing samples
  4. Add antibodies reactive with the same antigen (but different epitope)
  5. Wash and add chromogen -> measure OD
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7
Q

Competitive ELISA

A
  1. Unlabelled Ab are pre-incubated with bio samples containing antigen -> Ab-Ag complex formed
  2. Pipetted onto antigen coated ELISA plates
    (The more Ag present in bio sample -> the less Ab molecules remain free and available to bind antigen on ELISA plates)
  3. Apply enzyme-conjugated secondary Ab and chromogen
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8
Q

ELISPOT

A
  • Indirect immunoassay
  • Can detect cytokines (most often) secreted by individual cells. -> Determining number of positive cells even at a very low frequency
    1. The wells have either nitrocellulose or PVDF membrane bottom with captured Ab
    2. Pipetting live cells eg. lymphocytes into wells
    3. Incubation at 37 degrees for 1-2 days
    4. Wash cells out of wells
    5. The secreted molecules bound to the capture Ab are visualized by using a labeled secondary Ab and a substrate that will form an insoluble precipitate.
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9
Q

Immuno blot (Western blot)

A
  • Can determine the relative amount of a given protein within a biological sample using an antigen-specific primary Ab
    1. Different molecular weight proteins are separated by SDS page
    2. Blot (transfer) content of gel to a nitrocellulose or PVDF membrane
    3. Incubate membrane with a primary Ab specific to the desired.
    4. Incubation with secondary Ab (enzyme conjugated) that binds to primary
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10
Q

Radioimmunoassay (RIA)

A
  • Can determine conc. of eg. hormones

- A radioactively labeled antigen competes for Ab binding sites with the sample antigen

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11
Q

Immunoradiometric assay (IRMA)

A
  • Use monoclonal Ab bound to the inner surface of polystyrene tubes
  • Samples of patients incubated with radioactive Ab
  • Sample Ag will simultaneously bind to unlabelled Ab bound to the wall + radioactive Ab
  • Unbound radioactive Ab removed by washing
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12
Q

Immunocytochemistry

A
  • Can demonstrate specific proteins within cells/tissues using labeled Ab
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13
Q

Lateral flow test

A
  • Immerse a test strip into a biological sample and fluid migrates in the strip by capillarity
  • The sample is mixed with colored, specific Ab-coated microparticles (eg. latex)
  • Ag content of sample binds to colored microparticles and migrates along test strip
  • Accumulations of migrating colored microparticles
    1st spot: If it contains Ag of interest it will be dyed
    2nd spot: Anti-Ig Ab is dried onto the surface (serves as a positive internal control)
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14
Q

Serum electrophoresis

A
  • Used to identify missing/overproduced proteins
  • An electric field stimulates the movement of particles relative to a fluid
  • Detected: albumin, alpha globulins, HDL, beta globulin, gamma globulins
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15
Q

Decreased albumin in electrophoresis

A

Nephrosis syndrome, liver disease

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16
Q

Increased gamma-globulin in electrophoresis

A

Autoimmune diseases and infections

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17
Q

Increased total protein conc. detected in electrophoresis

A

Multiple myeloma

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18
Q

Immunoelectrophoresis

A
  • A special form og electrophoresis, for the separation and characterization of serum proteins
  • After electrophoresis, Ab are applied next to the separated proteins and immunoprecipitates are formed after a period of diffusion
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19
Q

Turbidimetry and nephelometry

A
  • Methods for determining the amount of cloudiness or turbidity in a solution based on measurment of the effect of this turbidity upon the transmission and scattering of light
  • Turbidity in a liquid is caused by the presence of finely divided suspended particles.
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20
Q

What can be diagnosed with flow cytometry?

A

To diagnose hematological disorders and immunodeficiencies

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21
Q

Steps of flow cytometry

A
  1. Sample prep: Blood from subject, separate mononuclear cells using a Ficoll gradient, stain with fluorescent Ab conjugates
  2. Instrument setup: Optimize fluorescence detector sensitivity
  3. Data acquisition: Pass stained cell suspension through laser beam
  4. Data analysis: Gate cell populations of interest
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22
Q

What can a flow cytometer tell us about a cell?

A
  • Its relative size (forward scatter)
  • Its relative granularity or internal complexity (side scatter)
  • Its relative fluorescence intensity
  • Time-dependency (kinetics) of these parameters
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23
Q

Which parameters can be measured by complement tests?

A
  • Conc. of individual complement factors
  • Conc. of regulatory factors
  • Activity of individual factors or overall activation
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24
Q

CH50 test

A
  • The demnominator of the serum dilution that lyses 50% of sheep RBCs in a test tube
    (eg. if we need 1/4 serum to lysis 50% of the RBCs then the CH50 will be 4) -> reciprocal
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25
What does a normal CH50 result tell us?
- Indicates that all complement factors (C1-9) are present | - The level of some however might be reduced a lot without affecting the CH50
26
What is more common, reduced or increased CH50 value?
Elevation, since complement proteins are also acute phase proteins.
27
Reduced CH50
- Production of a complement protein is not sufficient due to genetic mutations - Insufficient production (hepatic dysfunction, starvation) - Overconsumption in immunological disorders (eg. autoimmune disease, chronic infection)
28
Functional redundancy of cytokines
Many cytokines can have the same effect
29
Pleiotropy of cytokines
The same cytokines can have many different effects
30
What is a hapten?
A small molecule which, when combined with a larger carrier such as a protein, can elicit the production of antibodies which bind specifically to it (in the free or combined state)
31
What is an adjuvant?
An enhancer of the immune response to a given antigen by inducing costimulation
32
Types of vaccines used for active vaccination
1. Whole virus vaccines: live attenuated, inactivated, live recombinant 2. Subunit vaccine (Purified or recombinant antigen protein): synthetic peptide DNA
33
Why is there different vaccines for babies and elderly?
- Babies: Have a big naive B-cell pool, but few memory B-cells - Elderly: Have lots of memory B-cells and plasma cells but few naive B-cells
34
What is monoclonal antibody therapy?
A form of immunotherapy that uses monoclonal antibodies (mAb) to bind monospecifically to certain cells or proteins. This may then stimulate the patient's immune system to attack those cells.
35
Nomenclature of monoclonal Abs: Mab
Monnoclonal antibody
36
Nomenclature of monoclonal Abs: U
Human
37
Nomenclature of monoclonal Abs: O
Murine
38
Nomenclature of monoclonal Abs: Xi
Chimeric
39
Nomenclature of monoclonal Abs: Zu
Humanized
40
Nomenclature of monoclonal Abs: Li
Immunmodulating
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Nomenclature of monoclonal Abs: Tu
Antitumoral
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Nomenclature of monoclonal Abs: Ci
Cardiovascular diseases
43
CTLA4-Ig inhibits
CD28-B7 interactions
44
Effects H1 receptors
- Inc. mucous prod. - Inc. HR - Inc. vascular permeability - Constriction of SM in airways
45
Effect H2 receptors
- Vasodilation | - Inc. mucous secretion in airways
46
Effect H4 receptors
- Itch | - Recruitment of inflammatory cells by direct chemotaxis: eosinophils, DCs, mast cells
47
RAST test
- RadioAllergoSorbent test - A blood test that measures the levels of allergy IgE, when the patients blood is mixed with a series of allergens in laboratory
48
Difference between anaphylactic and anaphylactoid reactions
- Anaphylactic: Allergic or IgE mediated | - Anaphylactoid: Direct mast cell secretion (not a true allergic reaction)
49
Cause of urticaria and angioedema
- Urticaria: Due to edema of the dermis | - Angioedema: Due to edema of the subcutaneous tissue
50
Examples of type 2 hypersensitivity reactions caused by extrinsic antigen
1. Erythroblastosis fetalis: Mother produce Abs reacting with fetal RBCs 2. Transfusion reaction 3. Hyperacute allograft rejection 4. Drug induced hemolytic anemia
51
Examples of type 2 hypersensitivity reactions caused by intrinsic antigen (autoantibody)
1. Myasthenia gravis: Inhib. Ab against ach receptor | 2. Basedow disease: Stimulatory Ab against TSH-receptor
52
Direct Coombs test
Detects Abs on cell surface
53
Indirect Coombs test
Detects free Abs
54
Cryoglobulins can be detected in what autoimmune disease?
Frequent in Sjogrens syndrome
55
Cryoglobulins can be detected in what infections?
Most frequent in HCV
56
Cryoglobulins can be detected in what malignant tumors?
Most frequent in B-cell lymphoma
57
Antibodies detected in case of SLE
Antinuclear Abs (ANA), Anti-dsDNA, anti-Smith Ab, anti-chromatin Ab, anti-C1q Ab
58
Antibodies detected in case of myasthenia gravis
Anti-acetylcholine receptor Ab
59
Antibodies detected in case of Sjogrens syndrome
Anti-SSA and anti-SSB Ab
60
A person with Bechterews disease has this HLA
HLA-B27
61
A person with narcolepsy has this HLA
HLA-DR2
62
A person with IDDM has this HLA
HLA-DQ2
63
A person with rheumatoid arthritis has this HLA
HLA-DRB1
Immunology
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