Lecture_20_Objectives - detect reactive oxygen bacterial...

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Lecture 20 Objectives At the completion of the lecture and the assigned reading, the student should be able to: 1) Define the following terms [including what cell(s) the term applies to]: monoclonal antibody – Abs made by hybridoma cell line are all identical thus monoclonal antiserum – fluid comp of clotted blood from an immune individual that contains Abs against an Ag. Contains heterogeneous collection of Abs that bind the Ag 2) Given a suspected primary immunodeficiency, identify a laboratory test that would help to confirm or eliminate the suspected deficiency. (Example: Flow cytometric analysis for the absolute number of CD3 + CD8 + lymphocytes would be appropriate to test for a Tc deficiency due to MHC Class I deficiency.) Humoral Immunity defect – serum protein electrophoresis look for decreased/absence of gamma globulins, B cell count flow cytometry Tcell mediated immunity - Delayed hypersensitivity skin test, T cell count by flow cytometry, enzyme assays Phagocyte defect – Nitroblue tetrazolium (O2 sensitive dye) flow cytometry
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Unformatted text preview: detect reactive oxygen, bacterial killing assays, chemotaxis assays, leukocyte adhesion molecule expression by flow cytometry (CD11 a,b,c and CD18) neutrophil enzyme assays Complement defect – CH50 assay measures classical C’ path comp. EIA/ELISA detects C9 bound IgM coated microtiter wells serum C3 level 3) Explain the flow cytometry technique, including the reagent(s) that is/are used to label the individual cell population(s), and how that label is detected. Cell mixture labeled w/ fluorescent tagged monoclonal Abs specific for cell marker of interest. Cells suspended in saline ran thru cytometer, sep cells into single stream, activates the fluorochrome w/ a laser, detect fluorescent emission w/ detectors (also can simply do forward (cell size) and side (cell complexity) laser light scatter) B cell CD19, CD20 Total T cells CD3 also gamma/delta Tcells Th cells CD3+CD4 Tc cells CD3+CD8 NK cells CD16 OR CD56...
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