Immunodiagnosis

Question 1

LO 1. Describe the procedure used in serum protein electrophoresis and the underlying principles.

Answer 1

1. apply serum to gel and run (size, charge)
   2 stain the proteins
2. can and receive graph of data

Question 2

How are B cells counted?

Answer 2

use fluorescent mAbs specific for CD19 or CD20

Question 3

How are T cells counted?

Answer 3

use fluorescent mAbs specific for CD3, CD4, or CD8

Question 4

How does flow cytometry work?

Answer 4

machine counts cells by shooting laser light at single-file line of them; light scatter gives info about cell size and type

Question 5

How is the humoral system tested?

Answer 5

serum protein electrophoresis

Question 6

What is one issue with electrophoresis?

Answer 6

it’s not very sensitive to small abnormalities

Question 7

What is the order of the peaks on the serum protein electrophoresis, from left to right?

Answer 7

albumin (large), alpha1, alpha2, beta, gamma (tallish, wide)

Question 8

Electrophoresis can be done on which body samples?

Answer 8

plasma, urine, CSF

Question 9

What is the electrophoresis pattern look in MS?

Answer 9

oligoclonal peaks in the IgG region

Question 10

How would you measure individual Ig subclasses or subclasses?

Answer 10

single radial immunodiffusion

Question 11

What is the best overall test for Th1 activity?

Answer 11

skin test to common antigens

Question 12

What is a challenge DTH test?

Answer 12

use dinitrofluorobenzene to sensitize T cells- 98% will become sensitized if they’re normal

Question 13

Which mitogens can be used to stimulate T cells in mononuclear leukocyte preps?

Answer 13

PHA or Con A

Question 14

What are you looking for when you use mitogens to stimulate T cells?

Answer 14

proliferation or IL-2, IL-4, or IFN production; tells about T cell numbers and function

Question 15

What is a simple ELISA?

Answer 15

screen for antibodies present

Question 16

What disease can a simple ELISA be used to detect?

Answer 16

HIV

Question 17

How does a simple ELISA work?

Answer 17

1. attach antigen to the bottom of wells on a plate.
2. Add pts serum
3. If Ab is present, they will stick. Wash.
4. Add Ab to human IgG that has enzyme coupled to it that will change color. Wash.
5. Add color wash and look for reactions.

Question 18

What is a mitogen?

Answer 18

a plant protein that usu. binds sugar sequences on T cells

Question 19

What are antinuclear antibodies?

Answer 19

Antibodies against autoantigens in the nucleus

Question 20

How are antinuclear antibodies detected?

Answer 20

1. Grow human cells on a slide fixed with an agent that makes the cells’ plasma membranes permeable.
2. Add antibodies to penetrate to the cell’s interior.
3. Add patient’s serum. Wash.
4. Add fluorescein-labeled goat anti-human IgGs. Wash.
5. Examine slide under UV microscope for fluorescence and pattern

Question 21

What is the sandwich/capture ELISA used for?

Answer 21

to detect antibodies with at least 2 epitopes

Question 22

How does sandwich/capture ELISA work?

Answer 22

add antigen to wells that contain antibody; add another antibody that will fluoresce; view resulting color

Question 23

What is an example of a test that uses sandwich ELISA techniques?

Answer 23

rapid step test

Question 24

How is RF detected?

Answer 24

check for agglutination of latex particles coated with IgG because RF is IgM anti-IgG

Question 25

How are immune complexes detected?

Answer 25

put a sample in a cold environment (fridge)- if it precipitates after 1-7 days, it’s Type 3 mediated disease

Question 26

What is the final product of immunohistochemistry?

Answer 26

black or brown enzyme product in positive test

Question 27

What does a direct immunofluorescence test look for?

Answer 27

antigen

Question 28

What does an indirect immunofluorescence test look for?

Answer 28

antibody

Question 29

What is passive agglutination?

Answer 29

antigens are coupled to RBCs or latex beads; pt’s serum added; look for agglutination or not and titer.