Host Defense Clinical and Experimental Immunologic Assays II

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Host DefenseClinical and Experimental
Immunologic Assays II

• Domenick Kennedy
• Dept. Microbiology and Immunology
• dokennedy_at_lumc.edu

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Discussion Topics

• Common immunologic assays used for assessment of
  antibody response
• Production of Monoclonal Antibodies
• Immunofluorescence
• Flow Cytometry

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Monoclonal Antibody From a single clone of B
cells
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Clinically Used Monoclonal Abs
Infliximab Rheumatoid Arthritis, Crohns Disease,
Ulcerative Colitis Inhibits TNFa Rituximab Non-Hod
gkins Lymphoma Targets CD20 Trastuzumab Breast
Cancer Targets Her2/Neu (ErbB2)
receptor Adalimumab Rheumatoid Arthritis, Crohns
Disease, Ulcerative Colitis Inhibits TNFa
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Immunofluorescence
Mouse anti-Human Ig
Mouse anti-Human tumor antigen
Patient Serum
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Immunofluorescence Staining
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• Circumstances when we would want to identify a
  given cell population

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Peripheral Blood of HIV Patient
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anti-CD4, anti-CD8 antibodies
Analyze by Flow Cytometry
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CD4
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Flow Cytometry
Color
Granularity
Size
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SSC (Granularity)
FSC (Size)
http//www.nature.com/nprot/journal/v5/n7/fig_tab/
nprot.2010.88_F1.html
http//microvet.arizona.edu/Courses/MIC419/Secure/
CaseNoTap/flowfigsize.jpg
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Identification of Immune Cell Populations through
Flow Cytometry
CD8- Red CD4- Green
Color
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Measure CD4 and CD8 cells
CD4
Green
Red
HIV
-HIV
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Questions?
dokennedy_at_lumc.edu