Mcq Immuno

Dr. Tamma BMS 687 Molecular Immunology EXAM 1

Answer all the questions

1 Positive selection of B cells can be achieved by using following beads

a) Anti-CD3 coated beads

b) Anti-CD19 coated

c) Anti-CD14 coated beads

d) Staining with Trypan Blue

 

2 The following tests are used to measure cell proliferation

 

a) Trypan blue staining

b) SRBC resetting

c) MTT assay

d) All of the above

 

3 peripheral blood lymphocyte cell samples were placed in hemocytomer and cells were counted in 3

quadrants. Total Cell counts in 3 quadrants were 80. Following final calculations cell concentration was

2.6X10 6 /ml. What is the dilution factor? Please show how you calculated?

 

a) Dilution factor is 5

b) Dilution factor is 2

c) Dilution factor if 10

d) Dilution factor is 20

 

 

4. Following cell isolation and cell counting using hemocytometer, you obtained 6X10 6 /ml ( 6 million

cells per milliliter). Total volume you have is 1 ml.

You are asked to set up cell cultures as follows: 1) control cells without any stimulant, 2) cells with

stimulant A, 3) cells with stimulant B, 4) cells with both stimulant A and B. Total four conditions.

You are asked to use 1.5X10 6 /ml per condition to set up the experiment.

 

Therefore, you have to dilute the cells you obtained after counting (6X10 6 /ml).

To prepare 1.5X10 6 /ml per condition, how much medium do you have to add to your 6X10

6 /ml.

 

 

EXPLAIN: SHOW THE WORK OUT DETAILS

 

 

5 Trypan blue stains viable cells

a) True b) False

 

6 Show the correct order for MTT assay to assess cell proliferation

 

 

a) Prepare cells, incubate with MTT and culture for 3 days and read at 550-600

b) Prepare cells, incubate with detergent, culture, add MTT, read at 550-600

c) Culture cells for 72 hours, add MTT, add detergent, read at 550-600

d) Culture with MTT and detergent for 72 hours, read at 550-600

 

7 Peripheral blood lymphocyte cell samples were placed in hemocytomer and cells were counted.

Total Cell counts 120. Following multiplication with the dilution factor 10, cell concentration was

6X10 6 /ml. Cells were counted from how many quadrants?

 

a) 4

b) 3

c) 2

d) 1

 

 

8 The following are main ingredients in complete RPMI used for cell culture

a) 10% FCS; 1% Glutamine; 1% P/S (penicillin and streptomycin)

b) 1% FCS; 10% Glutamine; 12% P/S

c) 20% FCS; 1% Glutamine; 15% P/S

d) None of the above

 

 

9 cell counts were 1600; dilution factor was 2; final cell concentration was 3.2X10 7 /ml. how many

quadrants were used for cell counts?

 

a) 3

b) 4

c) 2

d) 1

 

10 Positive selection of T cells can be achieved by using the following antibody coated beads

a) Anti-CD3 coated beads

b) Anti-CD19 coated

c) Anti-CD14 coated beads

d) Staining with Trypan Blue

 

11 RBC and WBC can be separated based on density centrifugation by using

a) MTT

b) Ficol-Hypaque or Lymphoprep

c) Lympholysis

d) Trypan blue

 

 

 

 

 

12 Cell viability is assessed by using

a) MTT

b) FCS

c) Trypan Blue

d) Tetrazolium

 

13 A flowcytometer needs a combined system of the following

a) hemacytometer, buffer and cell counter

b) fluids, slides, and powerpack

c) optics, trypan blue and cell counter

d) electronics and hemacytometer, gel,

e) fluids, optics and electronics

 

14 The fluorochrome absorbs energy from the laser beam

A) True B) False

 

15 The fluorochrome releases the absorbed energy by: a) Vibration and heat dissipation

b) Emission of photons of a longer wavelength c) Fluids d) A and c e) A and b

 

16 Increased forward scatter is seen with

a) Neutrophils

b) Lymphocytes

c) RBC

d) Platelets

 

17 The relative levels of expression of cell surface molecules as measured by FACS is

a) forward scatter b) side scatter c) fluorescence intensity d) all of the above

 

 

18 Positive selection of the T cells can be achieved by cell sorting by using flowcytometry and what

antibody conjugated to fluorochrome is used?

a) Anti CD19 antibody conjugated to fluorochrome b) SRBC resetting c) Anti-CD3 antibody conjugated to fluorochrome d) Anti-CD14 antibody conjugated to fluorochrome e) All of the above

 

19 The isolation or purification of different cell populations with different surface antigens stained

with different fluorescent antibodies can be achieved by

a) immunohistochemistry b) enzyme linked immunosorbent assay c) immunoelectrophoresis d) fluorescene activated sorter.

 

20 CD25 is the receptor for

a) IL-12 b) IL-4 c) IL-2 d) IL-10

 

21 You stained the WBC with anti-CD3-FITC and anti-CD4-PE. Show the cells you expect to see in all

the four quadrants:

 

 

 

 

 

22 Positive selection of the B cells can be achieved by cell sorting by using flowcytometry and what

antibody conjugated to fluorochrome is used?

a) Anti CD19 antibody conjugated to fluorochrome

b) SRBC resetting c) Anti-CD3 antibody conjugated to fluorochrome d) Anti-CD14 antibody conjugated to fluorochrome e) All of the above

 

 

23 Positive selection of the Monocytes can be achieved by cell sorting by using flowcytometry and

what antibody conjugated to fluorochrome is used?

a) Anti CD19 antibody conjugated to fluorochrome b) SRBC resetting c) Anti-CD3 antibody conjugated to fluorochrome d) Anti-CD14 antibody conjugated to fluorochrome e) All of the above

 

24 You stained the WBC with anti-CD19-FITC and anti-CD3-PE. Show the cells you expect to see in all

the four quadrants:

 

 

 

 

 

25 You stained the WBC with anti-CD19-FITC and anti-CD8-PE. Show the cells you expect to see in

all the four quadrants:

 

 

 

 

 

26 You stained the WBC with anti-CD4-FITC and anti-CD25-PE. Show the cells you expect to see in

all the four quadrants:

 

 

 

 

 

 

 

27 You stained the cells from thymus with anti-CD4-FITC and anti-CD8-PE. Show the cells you expect to

see in all the four quadrants:

 

 

 

 

 

28 Annexin – FITC is used to identify cells in late apoptosis

a) true B) false

 

29. Propidium iodide is used to identify cells in late apoptosis

a) true B) false

 

30 Show Which histogram (close to Y axis or the one away from Y axis) represents increased

fluorescence intensity

 

 

 

31. Identify different cell Populations from the following slide – lysed whole

 

 

32 You are asked to set up an ELISA for the chemokine MIP-1. How do you design the experiment

(you can also use a second antibody)?

a) Coat the plate with MIP-1  wash, block with BSA  wash, add mouse

anti-MIP-1 antibody, incubate  wash, then add goat anti-mouse Ig- Alkaline Phosphatase enzyme (AP), incubate  wash, then add the substrate

(p-nitrophenol phosphate), incubate, look for color development.

 

b) Coat the plate with mouse anti-MIP-1 antibody  wash, add goat anti-mouse

Ig-AP,  add MIP 1,  then add the substrate

 

c) Coat the plate with goat anti-mouse Ig-AP, add MIP 1  add the substrate

d) Coat the plate with anti-MIP-1 antibody,  add MIP 1  add the substrate  add goat anti-mouse Ig-AP

 

e) All of the above 33 It has been reported by other researchers that a new intracellular protein (IPX) plays an important

role in T cell activation in humans. How do you design a western blot experiment to identify this IPX

protein???

a) Prepare cell lysate  run the polyacrylamide gel with the lysate  transfer the proteins to Nitrocellulose or PVDF membrane  block the membrane with BSA, incubate, wash,  add mouse anti-IPX antibody –Alkaline Phosphatase conjugated (AP), incubate, wash,  then add BCIP + NBT(substrate). Look for color development.

 

 

 

b) Prepare cell lysate  run the polyacrylamide gel with the lysate  transfer the proteins to Nitrocellulose or PVDF membrane  block the membrane with BSA, incubate, wash,  add mouse anti-IPX antibody alone, incubate, wash,  then add goat anti-mouse Ig-Alkaline Phosphatase (AP) enzyme,  incubate, wash,  then add BCIP + NBT. Look for color development.

 

c) Prepare cell lysate  run the polyacrylamide gel with the lysate  transfer the proteins to the nitrocellulose membrane  block the membrane with BSA, incubate, wash  add goat anti-mouse Ig-Alkaline Phosphatase (AP) enzyme, incubate, wash,  add the substrate.

 

d) Prepare cell lysate  run the polyacrylamide gel with the lysate  transfer the proteins to Nitrocellulose or PVDF membrane  block the membrane with BSA, incubate, wash,  add mouse anti-IPX antibody alone,  add substrate

 

e) a and b are correct

f) b and c are correct

g) all are correct

 

34 You immunized Goat with human IgD. You obtained serum ( which contains antibody to human

IgD) from the immunized goat. How do you purify goat anti-human IgD antibody?? Or what type of

affinity column you need to have??

a) Goat anti-human IgD antibody can be purified on human IgD-Sepharose (or agarose) column. Only anti-human IgD will bind to the column and can be eluted.

b) goat anti-human IgD can be purified on IgM-sepharose column c) goat anti-human IgD can be purified on IgG-sepharose column d) all are correct

35 You can purify different white cell populations using

a) Immunoprecipitation

b) Flow cytometry- cell soring

c) Specific antibody-coated magnetic beads

d) b and c

e) None of the above

 

 

36 You are asked to separate the following proteins in 10% SDS polyacrylamide gel (PAGE): IgM

(1000 kDa), Lysozyme (15 kDa), High density lipoprotein (485 kDa), Serum albumin (68 kDa),

and Transferrin (76 kDa). Choose the correct FASTEST order of migration of these proteins.

a) Lysozyme, High density lipoprotein, serum albumin, transferrin, IgM. b) Transferrin, serum albumin, IgM, high density lipoprotein, lysozyme. c) IgM, high density lipoprotein, transferrin, serum albumin, lysozyme,. d) Lysozyme, serum albumin, transferrin, high density lipoprotein, IgM.

 

37 The rate of migration of a protein through an SDS –polyacrylamide gel is influenced by all of the

following EXCEPT ONE, Which One??

a) size of the protein b) charge of the protein c) pore size of the gel d) The electric field.

 

38 Which of the following is the most frequently employed diagnostic laboratory technique for

detecting antigen antibody interactions using a Fluorochrome?

a) hemaglutination b) ELISA c) Immunofluorescence d) Immunoelectrophoresis.

 

39-40 Indicate True or false in the following:

a High molecular weight proteins can be separated easily by using high percentage of acrylamide-

bis acrymade.

b High molecular weight proteins can be separated easily by using low percentage of acrylamide-

bis acrymade.

c Low molecular weight proteins can be separated easily by using low percentage of acrylamide-bis

acrymade.

d Low molecular weight proteins can be separated easily by using high percentage of acrylamide-bis

acrymade.

 

 

 

 

 

41 which of the double diffusion profiles pictured below represents reaction of identity?

 

a)

 

 

b)

 

 

c)

 

 

a) none of the above