Laboratory Evaluation of the Cellular Immune System (RVSP) Flashcards
Solution used to obtain lymphocyte pellets
Ficoll hypaque density gradient soln
Identification of lymphocytes is useful in
Immunodeficiency diseases Lymphoproliferative malignancies Unexplained infectious diseases Monitoring of transplants Acquired immunologic abnormalities (AIDS)
ILUMA
3 ways to separate lymphocyte pellets
Nylon wool technique
Using monoclonal Abs (flow cytometry, lymphokwik)
Magnetic beads
T lymphocyte enumeration mtd that is manual
Rosette assay
Rgnt in rosette assay
Sheep’s RBCs
In rosette assay, sheep’s RBC’s attach to
CD2 of T cells
How many cells are ctd in Rosette assay?
200 cells
Formula for B cells
100 - rosettes
Tests for lab eval of Lymphocyte function
Transformation assays
- mitogen-induced blastogenesis
- mixed lymphocyte culture
Cytotoxicity assay
Mitogen-induced blastogenesis (MIB) is an assessment of
Proliferative capacity of lymphocytes
Incubation period of mitogens in mitogen-induced blastogenesis (MIB)
3-4 days
Incubation period of antigens in mitogen-induced blastogenesis
5-10 days
Radiolabeled DNA nucleoside used in mitogen-induced blastogenesis
Tritiated thymidine
Methods for manual lymphocyte enumeration
Hemocytometry
Rosette assay
In MIB, after incubation, the harvest cells are transferred to
Disks of filter paper
In MIB, the disks of filter paper are placed in
Scintillation counter
MIB method where the response of the entire cell population is measured
Bulk Assay Mtd
Mitogens for T cells
Phytohemagglutinin Concanavalin A (Con A)
Mitogens for B cells
Staphylococcal protein A
Pokeweed mitogen