Term
| What cells are considered peripheral blood mononuclear cells (PBMCs)? What chemical solution has the same specific gravity as PBMCs? |
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Definition
T cells, B cells, NK cells, monocytes
Ficoll-Hypaque solution (used to separate PBMCs from blood) |
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Term
| What is the purpose of the lymphocyte proliferation assay? |
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Definition
| To detect if lymphocytes will react and clonally expand to an antigen. Test for increased radioactivity (from thymidine) |
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Term
| What is the purpose of the ELISPOT assay? |
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Definition
| Detect number of T cells/B cells that are secreting cytokine/antibody |
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Term
| Chromium (51-Cr) is used as a marker to detect what? |
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Definition
| CTL-mediated cytotoxicity - how effective they are at killing the target cells marked with Cr. |
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Term
| What are some in vivo immune responses where in vitro lymphocyte proliferation assays can be used as a model? |
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Definition
1) Delayed type hypersensitivity (DTH)
- Memory cells after initial exposure to antigen
2) Transplant rejection
- expose recipient's lympoctes with MHCs from donor (mixed lymphocyte response)
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Term
| In flow cytometry, what does side scatter measure? forward scatter? |
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Definition
side scatter = complexity (granules)
forward scatter = size |
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Term
| What is the purpose of flow cytometry? |
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Definition
| To determine the types of markers and receptors on the surface of a cell |
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Term
| What do you call the results of flow cytometry when only one fluorescent marker is used? what about when two or more markers are used? |
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Definition
One marker - results seen as histogram
two or more marker - scattergram |
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Term
| What is a normal CD4/CD8 ratio? |
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Definition
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Term
| What does a precipitin reaction look for? what percentage of antibody to antigen must there be for optimum reaction? |
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Definition
A precipitin reaction looks for a lattice of antibody-antigen (immune complex) forming a precipitate. For this reaction, antigen is in soluble form.
There should be equivalent concentrations of antigen and antibody to form a precipitate. |
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Term
| What are some applications of antigen-antibody reactions? |
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Definition
- Blood typing
- Coombs direct/indirect test (Rh factor)
- Measure rheumatoid factor
- C reactive peptide
- antibodies to treponema pallidum
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Term
| What are some applications for ELISA? |
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Definition
| measure antibodies to virus (ie HIV or hepatitis) |
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Term
| What is the purpose of immunofluorescence/immunohistochemistry? What is one of its applications? |
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Definition
Purpose: look at exact location and type of cell in situ
Applications: study proteins in cancers. |
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Term
| For a positive western-blot of HIV, what markers must be detected? |
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Definition
| p24 and either gp41, gp120 or gp160 |
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Term
| True or false. A positive ELISA for HIV antibodies is confirmation for diagnosis of HIV. |
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Definition
False.
a positive ELISA must be followed with a western-blot to confirm. |
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Term
| What is FACS and for what diseases can it be diagnostic? |
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Definition
Fluorescence-Activated Cell Sorting
- Uses flow cytometry identify cell with cell markers
- provides relative and absolute counts of cell types in blood
Dx: infections, neutropenia, leukopenia, or other immunodeficiencies |
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Term
| How can you measure B-cell funcion in vivo and in vitro? |
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Definition
In vivo:
- serum Ig levels
- specific antibody titers
In vitro:
- mitogen-induced Ab production |
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Term
| How can you measure T-cell funcion in vivo and in vitro? |
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Definition
In vivo:
- DTH response to skin test
In vitro:
- T-cell proliferation in response to mitogen (i.e. phytohemagglutinin, PHA) |
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