Laboratory Evaluation of the Cellular Immune System (RVSP) Flashcards

0
Q

Solution used to obtain lymphocyte pellets

A

Ficoll hypaque density gradient soln

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1
Q

Identification of lymphocytes is useful in

A
Immunodeficiency diseases
Lymphoproliferative malignancies
Unexplained infectious diseases
Monitoring of transplants
Acquired immunologic abnormalities (AIDS)

ILUMA

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2
Q

3 ways to separate lymphocyte pellets

A

Nylon wool technique
Using monoclonal Abs (flow cytometry, lymphokwik)
Magnetic beads

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3
Q

T lymphocyte enumeration mtd that is manual

A

Rosette assay

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4
Q

Rgnt in rosette assay

A

Sheep’s RBCs

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5
Q

In rosette assay, sheep’s RBC’s attach to

A

CD2 of T cells

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6
Q

How many cells are ctd in Rosette assay?

A

200 cells

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7
Q

Formula for B cells

A

100 - rosettes

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8
Q

Tests for lab eval of Lymphocyte function

A

Transformation assays

  • mitogen-induced blastogenesis
  • mixed lymphocyte culture

Cytotoxicity assay

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9
Q

Mitogen-induced blastogenesis (MIB) is an assessment of

A

Proliferative capacity of lymphocytes

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10
Q

Incubation period of mitogens in mitogen-induced blastogenesis (MIB)

A

3-4 days

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11
Q

Incubation period of antigens in mitogen-induced blastogenesis

A

5-10 days

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12
Q

Radiolabeled DNA nucleoside used in mitogen-induced blastogenesis

A

Tritiated thymidine

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13
Q

Methods for manual lymphocyte enumeration

A

Hemocytometry

Rosette assay

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14
Q

In MIB, after incubation, the harvest cells are transferred to

A

Disks of filter paper

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15
Q

In MIB, the disks of filter paper are placed in

A

Scintillation counter

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16
Q

MIB method where the response of the entire cell population is measured

A

Bulk Assay Mtd

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17
Q

Mitogens for T cells

A
Phytohemagglutinin
Concanavalin A (Con A)
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18
Q

Mitogens for B cells

A

Staphylococcal protein A

Pokeweed mitogen

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19
Q

Mixed lymphocyte assay (MLC) is a special type of lymphocyte __________ assay

A

Stimulation

20
Q

Major determinant in MLC

A

D locus of HLA system

21
Q

Test used to assess the ability of histoincompatible lymphocytes from one indiv to stimulate lymphocytes of another indiv

A

Mixed lymphocyte culture (MLC)

22
Q

In MLC, ___________ cells are mixed with _____________

A

Irradiated stimulator cells

Irradiated responder cells

23
Q

Used in MLC for incubation

A

Cell culture

24
Q

Used in MLC to determine degree of stimulation

A

Scintillation counter

25
Q

Test used in the selection of compatible donor for a BM transplant

A

MLC

26
Q

Endpoint of cytotoxicity assay

A

Cell death

27
Q

Cell death may be due to

A
  • Complement activity (complement-mediated cytotoxicity)

- Direct effect of one cell to another cell (cell-mediated cytotoxicity)

28
Q

Target cell lysis is determined by the release of ________ from the target cell or incorporation of a _________

A
Chromium
Vital dye (eosin, trypan blue)
29
Q

Substances used in Chromium-release assay for stimulation

A

Phytogemagglutinin

Labeled 51Cr

30
Q

True/False. Chromium-release assay uses PBMC

A

True

31
Q

Represents unlysed cells in chromium-release assay

A

Radioactivity in the cells at the end of the experiment

32
Q

Std technique for detection of Ag-Ab interaction on a cell surface

A

Dye Viability Exclusion Assay

33
Q

Explain the assay for neutrophilic activation

A

Stimulatory substances bind to surface receptors to illicit metabolic and structural changes that result to phagocytosis, oxidative burst, locomotion, and degranulation

34
Q

Assay that uses particles that are visible within the cell and can be counted after ingestion

A

Assay of phagocytosis and endocytosis

35
Q

Target cells for the assay of phagocytosis and endocytosis

A

Neutrophil

Monocyte

36
Q

In the assay of phagocytosis and endocytosis, target cells are incubated with

A

Yeast

37
Q

Stain used in the assay of phagocytosis and endocytosis

A

Giemsa

38
Q

Number of cells counted in the assay of phagocytosis and endocytosis

A

200 cells

39
Q

Increase in oxidative metab during phagocytosis

A

Oxidative burst

40
Q

Test used in the assay of enzymatic and intracellular killing

A

Nitro Blue-Tetrazolium test (NBT)

41
Q

Formed when there is a nmal oxidizing environment

A

Formazan

42
Q

Color of NBT in oxidized form

A

Yellow

43
Q

Color of formazan

A

Deep blue

44
Q

True/False. NBT Test is a quantitative assay

A

False. It is a SEMIQUANTITATIVE assay

45
Q

Disease where the PMNLs will not turn blue in the NBT test

A

Chronic Granulomatous Disease

46
Q

Tests the phagocytic function of polymorphonuclear leukocytes (PMNLs)

A

Nitro Blue-tetrazolium test

47
Q

Opsonin in the serum can be detected by incubating serum with __________ and ____________

A

Normal neutrophils

Yeast particles

48
Q

Assay of membrane permeability uses _____________

A

Fluorescein diacetate