Host Defense Clinical and Experimental Immunologic Assays II Domenick Kennedy Dept. Microbiology and Immunology

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Presentation transcript:

Host Defense Clinical and Experimental Immunologic Assays II Domenick Kennedy Dept. Microbiology and Immunology

Discussion Topics Common immunologic assays used for assessment of antibody response: – Production of Monoclonal Antibodies – Immunofluorescence – Flow Cytometry

Monoclonal Antibody – From a single clone of B cells

Clinically Used Monoclonal Abs Infliximab Rheumatoid Arthritis, Crohn’s Disease, Ulcerative Colitis Inhibits TNFα Rituximab Non-Hodgkin’s Lymphoma Targets CD20 Trastuzumab Breast Cancer Targets Her2/Neu (ErbB2) receptor Adalimumab Rheumatoid Arthritis, Crohn’s Disease, Ulcerative Colitis Inhibits TNFα

Ig Immunofluorescence Mouse anti-Human tumor antigen Patient Serum Mouse anti-Human Ig

Immunofluorescence Staining

Circumstances when we would want to identify a given cell population:

Peripheral Blood of HIV Patient

CD4 + CD8 + CD4 + CD8 + CD4 + + anti-CD4, anti-CD8 antibodies Analyze by Flow Cytometry

CD4 + CD8 + CD4 + CD8 + CD4 +

Flow Cytometry Color Size Granularity

SSC (Granularity) FSC (Size)

Identification of Immune Cell Populations through Flow Cytometry Color CD8- Red CD4- Green

Measure CD4+ and CD8+ cells +HIV-HIV Green Red CD8 + CD4 +

Questions?