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bio 220 [exam #4] > Chapter 18 - Practical Applications of Immunology > Flashcards

Chapter 18 - Practical Applications of Immunology Flashcards

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1
Q

What are some comon practical applications (uses) of immunology?

A
  • Vaccination: inject Ag to trigger immune repsonse (make Ab)
  • Serological Testing: test PT blood (serum) and look for Ag OR Ab (IgM) = timely and accurate diagnosis
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2
Q

Briefly describe how Vaccination works in regards to primary and secondary immune responses

A
  1. Injection of Ag will provoke a primary immune reponse -> formation of memory B cells and Ab (after clonal sel and exp)
  2. Will produce rapid, intense secondary immune response
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3
Q

A substance (Ag) that stimulates the immune system is known as a…

A

immunogen

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4
Q

This is described as protection for most of the population

A

Herd Immunity

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5
Q

Name 5 desirable characteristics for an ideal vaccine. Describe each

A

SEOOA

1. Vaccine Safety

  • Vaccine must never be pathogenic/toxigenic

2. Vaccine Effectiveness

  • Vaccine must be a strong immunogen (ex. protein) and stimulate immune system

3. Vaccine with Oral Administration

  • Orally (painless) > injectable (painful)

4. Vaccine Require One Injection

  • One injection/combination vaccine preferred (PT less likely to come back if req. multiple injection)

5. Vaccine is Affordable/Easily Accessible

  • Availiable to most people @ resonable cost
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6
Q

Name the different Types of Vaccines (6)

A
  1. Live, attenuated vaccine
  2. Inactivated, killed vaccine
  3. Toxoid vaccines
  4. Conjugated vaccines
  5. Subunit vaccine
  6. mRNA (nucleic acid) vaccine
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7
Q

Compare these Types of Vaccines: Live, attenuated vaccine vs. Inactivated, killed vaccine

Include examples of DZ that use these types of vaccine

A

Live, attenuated vaccine

  • Weakened pathogen
  • Closely mimic actual infection (bc inj. Ag is still alive)
  • Ex: MMR, Smallpox

Inactivated, killed vaccine

  • Killed pathogen
  • Safer (compared to live vaccines)
  • Ex: Influenza, Polio
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8
Q

Describe: Toxoid vaccines, Conjugated vaccines, Subunit vaccines, mRNA (nucleic acid) vaccines

A

Toxoid vaccines

  • inactivated toxin (make Ab against toxoid = antitoxin)
  • immune response will target toxin
  • Ex: Td vaccine

Conjugated vaccines

  • main vaccine component is combined with a strong immunogen (protein)
  • Ex: Pneumonia vaccine

Subunit vaccines

  • Use antigenic fragments (piece of pathogen CW) to stimulate immune response
  • Ex: Petussis vaccine; Hep. B vaccine

mRNA vaccines

  • inject only bacterial/viral nucleic acid (NOT live pathogen)
    -> inject mRNA into host cell -> make protein Ags -> stimulate immune response to make Ab
  • Ex: Covid vaccine
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9
Q

What part of the blood sample is being tested?

A

serum (no CF; top part)

YOU ARE NOT SAMPLING THE PLASMA (has CF)

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10
Q

When using serological testing, what are we looking for? (Hint: Theres two things we could look for)

A

Antigen or Antibody (IgM)

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11
Q

This is the study or diagnositc examination of blood serum (to see how the immune system response to pathogens or substances introduced into the body)

A

Serology

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12
Q

Name 5 desirable criteria for an ideal serological test

A
  • High specificity/accuracy
  • High sensitivity
  • East set-up & interpretation
  • High Throughput
  • Affordable cost and Easy Access
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13
Q

Describe the criteria for an ideal serological test

A

High specificity/accuracy

  • Unambigious (one interpretation)
  • Reliable (100% accuracy)
  • Ex: Pregnancy test; HIV test; athlete drug doping test

High sensitivity

  • Uses small sample size = detect small quantities Ag or Ab

East set-up & interpretation

  • test construction simple
  • results easy to read (ex: color change; + or - symbol)

High Throughput

  • Many tests carried out at once (bc machine automation) = fast results

Affordable cost and Easy Access

  • Avalibale to most people @ reasonable cost
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14
Q

Define: Throughput

A

ability to stimultaneously test hundreds/thousands of samples

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15
Q

There are two types of serological tests. Name them and describe what you are looking for

A

Two Tests:

Direct Serological Test

  • Look for Ag (pathogen)

Indirect Serological Test

  • Look for Ab (IgM)
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16
Q

An antigen or antibody is provided either by the _____ sample or the ______ ______

A

patient
lab test

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17
Q

Understand what a serological test will have and what the PT serum sample would have to provide

A

If serological test has Ag, the PT serum sample will provide the Ab

If serological test has the Ab, the PT serum sample wil provide the Ag

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18
Q

Name (6) common examples of serological testing

A
  • Precipitation Reactions
  • Agglutination Reactions
  • Neutralization Reactions
  • Fluorescent-Antibody Technique
  • Complement-Fixation Reactions
  • ELISA
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19
Q

Describe this example of serological testing: Precipitation Reactions
Include: Types of testing avaliable, Component involved (Ag/Ab), Interpretation of Results (What does the PT/Lab provide, Know what a positive/negative test looks like), Clinical Applications

A

Types of Testing Avaliable

  • Direct Tests
  • Indirect Tests

Component involved (Ag/Ab)

  • Ag: soluble antigens (not attached to pathogen)
  • Ab: IgG/IgM

Interpretation of Results

POSITIVE Direct Test:

  • PT (serum sample) provide: Ag
  • Lab provide: Ab
  • PT provide soluble Ag -> Ab (lab) binds to Ag (PT) -> immune complex forms -> precipitan ring at equivalence zone = PT HAS DZ

POSITIVE Indirect Test:

  • PT (serum sample) provide: Ab
  • Lab provide: Ag
  • PT provide Ab -> Ab (PT) binds to Ag (lab) -> immune complex forms -> precipitan ring at equivalence zone = PT has DZ

Clinical Applications

  • VDRL Test
    -> Blood test for Syphilis only
    -> Look for PT Ab (against T. pallidum) = Indirect test
  • Lancefield Classification of Streptococcal Species
    -> Classifies streptococci into groups from A-K and H-U (ex: GAS, GBS, GCS)
    -> Helps distinguish between different species of Genus Streptococcus
20
Q

In regards to precipitation reactions, just by looking at a test tube, how would you tell a PT has a disease?

A

immune complexes form at equivalence zone, where a precipitan ring is present.
precipital ring = cloudy line (immune complex present) where 1:1 ration of Ag to Ab

21
Q

Describe this example of serological testing: Agglutination Reactions
Include: Types of testing avaliable, Component involved (Ag/Ab), Interpretation of Results (What does the PT/Lab provide, Know what a positive/negative test looks like), Clinical Applications

A

Types of Testing Avaliable

  • Direct Tests
  • Indirect Tests

Component involved (Ag/Ab)

  • Ag: particulate antigens (attached to pathogen)
  • Ab: IgM

Interpretation of Results

POSITIVE Direct Test:

  • PT (serum sample) provide: Ag
  • Lab provide: Ab (IgM)
  • Ag and IgM present = immune complex = clumping = POSITIVE TEST

POSITIVE Indirect Test:

  • PT (serum sample) provide: Ab
  • Lab provide: Ag
  • Ag cand IgM present = immune complex = clumping = POSITIVE TEST

Clinical Applications

  • Hemaggulitnation
    -> For blood typing; Agglutination of RBC surface Ag & complimentary Ab
  • Antibody Quantitation
    -> measuring Ab titer
    -> Differentiate bw primary (high IgM) & secondary (high IgG) exposure
22
Q

Know how to tell what blood type you have in regards to hemagglutination.

Ex: If you have two wells, one with anti-A Ab and one with anti-B Ab, what would you observe you placed a PT serum sample who has Type A blood into these two wells (hint: which wells would clump)?

A

Type A:

  • Anti-A well: clumping
  • Anti-B well: x

Type B:

  • Anti-A well: x
  • Anti-B well: clumping

Type AB:

  • Anti-A well: clumping
  • Anti-B well: clumping

Type O:

  • Anti-A well: x
  • Anti-B well: x
23
Q

Describe this example of serological testing: Neutralization Reactions
Include: Types of testing avaliable, Component involved (Ag/Ab), Interpretation of Results (What does the PT/Lab provide, Know what a positive/negative test looks like), Clinical Applications

A

Types of Testing Avaliable

  • Indirect Tests

Component involved (Ag/Ab)

  • Ag: Toxin/Virus
  • Ab: Antitoxin/Antivirals
  • Cell Indicator

Interpretation of Results

Toxin Neutralization Test: + Indirect Test:

  • PT (serum sample) provide: Ab
  • Lab provide: Ag (toxin) & cell indicator
  • PT serum sample (has Ab) -> lab adds Ag and cell indicator -> PT Ab and Lab Ag form immune complex -> toxin is neutralized -> cell indicator is UNDAMAGED -> PT HAS DZ
  • IF NEGATIVE: PT serum sample does NOT have Ab -> Lab adds Ag and cell indicator -> no Ab (PT) present -> toxin not neutralized -> toxin attack cell indicator -> cell indicator damage -> PT DOES NOT HAVE DZ

Viral Hemaggulitnation Inhibition Test: + Indirect Test:

  • PT (serum sample) provide: Ab
  • Lab provide: Ag (virus) & cell indicator (RBC)
  • Pt serum sample (has Ab) -> lab adds Ag (virus) and cell indicator (RBC) -> PT Ab bind to lab Ag (virus) -> virus neutralized -> hemagglutinatoin inhibited -> NO CLUMPING -> PT HAS DZ
  • IF NEGATIVE: PT serum sample (NO Ab) -> lab adds Ag (virus) and cell indicator (RBC) -> no Ab (PT) present-> virus not neutralized -> hemagglutinatoin of RBC ->YES CLUMPING -> PT DOES NOT HAVE DZ

Clinical Applications

  • DX of bacterial disease (toxin production)
  • DX of viral disease
24
Q

Describe this example of serological testing: Fluorescent Antibody Reactions
Include: Types of testing avaliable, Component involved (Ag/Ab), Interpretation of Results (What does the PT/Lab provide, Know what a positive/negative test looks like), Clinical Applications

A

Types of Testing Avaliable

  • Direct Tests
  • Indirect Tests

Component involved (Ag/Ab)

  • Ag: Streptococci or T. Pallidum
  • Ab: Fluorescent Ab or Anti-human Ab

Interpretation of Results

POSITIVE Direct Florescent-Antibody (FA) Test:

  • PT (clinical/throat sample) provides: Group A streptococci (Ag)
  • Lab provides: Fluorescent-Dyed Ab
  • PT sample (has Group A streptococci Ag) -> lab adds fluorescent-dyed Ab -> Immune complex forms -> Fluorescent Streptococci (chains)-> POSITIVE, PT has DZ
  • IF NEGATIVE: No fluorescent streptococci (chains)

POSITIVE Indirect Florescent-Antibody (FA) Test:

  • PT (serum sample) provides: Ab
  • Lab provide: T. pallidum (Syphilis) & Fluorescent Antihuman Ab (Ab that binds to human Ab)
  • PT sample (has Ab against T. Pallidum) -> Lab adds Ag (T. pallidum) and Fluorescent Anti-human Ab -> Immune complex forms -> Fluorescent Antihuman Ab binds to PT Ab of immune complex-> fluorescent T. pallidum (spiral) -> POSITIVE, PT has DZ
  • IF NEGATIVE: No flourescent T. pallidum (spiral)

Clinical Applications

  • Identify GAS from throat sample (direct FA test)
    -> Ex: diagnose strep throat/scarlet fever by looking for streptococcus pyogenes
  • See if T. pallidum from PT blood is producing Ab (indirect FA test)
    -> Used to diagnose Syphilis
25
Q

What types of serological tests (reactions) can be used to diagnose Syphilis? Are these direct or indirect tests?

A

VDRL Test (Precipitation RXN)
- Indirect test

Florescent-Antibody Test (Flourescnet Antibody Technique)
- Indirect Test

26
Q

Describe this example of serological testing: Complement Fixation Reactions
Include: Types of testing avaliable, ALL Component involved, Interpretation of Results (What does the PT/Lab provide, Know what a positive/negative test looks like), Clinical Applications, MISC

A

Types of Testing Avaliable

  • Indirect Tests

Component involved

  • Antibody (PT)
  • Antigen (Lab)
  • Complement Protein (Lab)
  • Sheep RBC (Lab)
  • Antibody to sheep RBC (Lab)

Interpretation of Results

POSITIVE Indirect Complement-fixation Test:

  • PT (serum sample) provides: Ab
  • Lab provide: Ag & complement
  • YES PT has Ab -> Immune complex forms -> complement binds/fixed to stem region -> complement fixation (all complement used up/no more CP available)
  • Add Sheep RBC & Ab to RBC -> NO hemolysis -> YES PT has DZ

NEGATIVE Indirect Complement-fixation Test:

  • PT (serum sample) provides: No Ab
  • Lab provide: Ag, complement, Sheep RBC, Anti RBC Ab
  • No PT Ab = No immune complex forms -> No complement fixation (all complement available)
  • Add Sheep RBC & (Ab to sheep RBC) -> complement binds to Sheep RBC Ab -> YES hemolysis -> red color -> NO DZ

Clinical Applications

  • DX of bacterial DZ
  • Dx of viral DZ

MISC

  • Can detect small amounts of Ab = Highly SENSITIVE test
27
Q

Name the two stages for the Complement Fixation RXN Test

What do these stages tell us?
Which parts (based on exam figures) happen in each stage?

A

1. Compliment fixation stage

  • Tells us if complement gets fixed/used up
  • Part 1 and 2

2. Indicator stage

  • Tell us if PT is positive/negative for DZ
  • Part 3 and 4
28
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 1 for a POSITIVE CF Test

A
  • An Ag and inactivated complement are provided by the lab
  • These are added to the serum sample containing the PT Ab which are specific for the Ag
  • An immune complex will form consisting of the labs Ag and the PT Ab
  • This is part of the “Compliment Fixation Stage” of the test
29
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 2 for a POSITIVE CF Test

A
  • As a result of immune complex formation, compliment gets activated and compliment fixation occurs (all availiable complement binds to antibody stem region of the immune complex)
  • Since all of the complement got used up, no more compliment will be availiable to bind to future immune complex formations
  • This is part of the “Compliment Fixation Stage” of the test
30
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 3 for a POSITIVE CF Test

A
  • A sheep RBC and Ab against sheep RBC are provided by the lab and are added to the PT serum sample
  • The sheep RBC is used as a test indicator
  • An immune complex will form between the sheep RBC and the Ab against sheep RBC
  • This is part of the “Indicator Stage” of the test
31
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 4 for a POSITIVE CF Test

A
  • No hemolysis of sheep RBC occurs because compliment did not bind to the Ab against sheep RBC
  • Compliment did not bind because it was all used-up and already fixed (complement fixation) in the earlier step to the first immune complex. Therefore, complement was no longer availiable
  • This part is the “Indicator Stage” of the test
32
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 1 for a NEGATIVE CF Test

A
  • An antigen and inactivated complement are provided by the lab
  • These are added to the serum sample that does NOT contain the PT antibodies which would have been specific for the antigen
  • NO immune complex will forms because the PT serum sample has no Ab against the labs Ag
  • This is part of the “Compliment Fixation Stage” of the test
33
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 2 for a NEGATIVE CF Test

A
  • As a result of NO immune complex formation, compliment does NOT get activated and compliment fixation does NOT occur (compliment does not bind to Ab stem region of the immune complex)
  • Since NO complement got used up (since it did not bind to Ab stem region) all of the compliment will be availiable to bind to future immune complex formations
  • This is part of the “Compliment Fixation Stage” of the test
34
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 3 for a NEGATIVE CF Test

A
  • A sheep RBC and Ab against sheep RBC are provided by the lab and are added to the PT serum sample
  • The sheep RBC is used as a test indicator
  • An immune complex will form between the sheep RBC and the Ab against sheep RBC
  • This is part of the “Indicator Stage” of the test
35
Q

In regards to the exam figure regarding Compliment-Fixation Reaction Test, describe what is going on in Stage 4 for a NEGATIVE CF Test

A
  • YES hemolysis of sheep RBC occurs because compliment was activated and compliment fixation now occurs (all availiable complement binds to Ab stem region of immune complex)
  • Compliment binds to Ab against sheep RBC because it was not used up/not fixed (no compliment fixation) in the earlier step because no immune complex formed during the Compliment Fixation Stage of the test. Therefore, compliment was availiable
  • Hemolysis of sheep RBC indicates a negative test for the patients Ab, and thus, negative for the disease
  • This part is the “Indicator Stage” of the test
36
Q

Describe this example of serological testing: ELISA
Include: Types of testing avaliable, ALL Component involved (Ag/Ab), Interpretation of Results (What does the PT/Lab provide, Know what a positive/negative test looks like), Clinical Applications, MISC

A

Types of Testing Avaliable

  • Direct Test
  • Indirect Tests

Component involved

Direct ELISA:

  • (HIV) Ag (PT)
  • Adsorbed Ab (Lab)
  • Enzyme-Linked Ab (Lab)
  • Substrate (Lab)

Indirect ELISA:

  • (HIV) Adsorbed Ag (Lab)
  • Ab (PT)
  • Enzyme-Linked Antihuman Ab (Lab)
  • Substrate (Lab)

Direct ELISA (Pregnancy Test):

  • hCG hormone from urine sample (PT)
  • 2 Monoclonal Ab for hCG (test)

Interpretation of Results

POSITIVE Direct Test HIV Testing

  • PT (serum sample) provides: HIV Ag
  • Lab Provides: Adsorbed Ab, Enzyme-linked Ab, substrate
  • Lab Ab adsorbed to well -> immune complex forms -> Enz-linked binds to Ag forming sandwich -> enz substrate added -> RXNS occur color change to purple

POSITIVE Indirect Test HIV Testing

  • PT (serum sample) provides: Ab
  • Lab Provides: Adsorbed HIV Ag, Enzyme-linked Anti-human Ab, substrate
  • Lab HIV Ag adsorbed to well -> PT serum sample added-> immune complex forms -> Enz-linked Antihuman Ab binds to Ab-> enz substrate added -> RXNS occur color change to purple

POSITIVE Direct Test At Home Pregnancy Test

  • PT (urine sample) provides: hCG hormone
  • Test provides: Captured Ab bound to substrate
  • Urine sample added -> both monoclonal Ab bind to hCG hormone-> sandwich form -> color change/line forms

Clinical Applications

  • Home pregnancy test (direct ELISA)
  • Drug testing (direct ELISA)
  • HIV testing (direct ELISA)
  • HIV Testing after seroconversion (indirect ELISA)

MISC

  • High specificity
  • High sensitivity
37
Q

Name serological tests that are have high sensitivity (4), intermediate sensititivty (2), and low sensitivity (1)

A

High Sensitivity

  • Radioimmunoassay
  • PCR
  • Compliment-Fixation Test
  • ELISA

Intermediate Sensitivity

  • Agglutination RXN
  • Neutralization RXN

Low Sensitivity

  • Precipitation RXN
38
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 1 for a POSITIVE DIRECT ELISA Test

A
  • Antibody (against Ag/HIV) from the lab is adsorbed to well
39
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 2 for a POSITIVE DIRECT ELISA Test

A
  • PT sample (containing HIV) is added to the well containing the adsorbed Ab against HIV (from lab)
  • An immune complex will form between PT HIV and the adsorbed Anti-HIV Ab
40
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 3 for a POSITIVE DIRECT ELISA Test

A
  • An enzyme-linked Ab (specific to HIV) from the lab is added into a well containing an immune complex between the adsorbed Ab from the lab and the PT HIV.
  • The enzyme-linked Ab binds to the PT HIV (Ag) which is a part of an immune complex
  • This orientation/formation is known as a sandwich.
41
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 4 for a POSITIVE DIRECT ELISA Test

A
  • The enzyme’s substrate from the lab is added
  • The substrate will bind to the enzymes of the enzyme-linked antibody and this reaction produces a product that results in a visible color change
  • This indicates that the PT is POSITIVE for HIV.
42
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 1 for a POSITIVE INDIRECT ELISA Test

A
  • HIV (Ag) from the lab is adsorbed to the well
43
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 2 for a POSITIVE INDIRECT ELISA Test

A
  • PT sample (containing Ab against HIV) is added to the well containing the adsorbed HIV (Ag) from the lab
  • An immune complex will form between the PT Anti-HIV Ab and the absorbed HIV (Ag)
44
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 3 for a POSITIVE INDIRECT ELISA Test

A
  • An enzyme-linked anti-human Ab from the lab is added into a well containing an immune complex between the adsorbed HIV (from lab) and the PT Ab against HIV.
  • The enzyme-linked anti-human Ab binds to the PT Ab against HIV that is a part of an immune complex

(NO SANDWICH FORMATION OCCURS IN INDIRECT ELISA)

45
Q

In regards to the exam figure regarding ELISA Testing, describe what is going on in Step 4 for a POSITIVE INDIRECT ELISA Test

A
  • The enzyme’s substrate from the lab is added
  • The substrate will bind to the enzymes of the enzyme-linked anti-human Ab and this reaction produces a product that results in a visible color change
  • This indicates that the PT is POSITIVE for HIV.

bio 220 [exam #4] (8 decks)

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