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Radioimmunoassay, enzymoimmunoassay - principles, practical use.

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Presentation on theme: "Radioimmunoassay, enzymoimmunoassay - principles, practical use."— Presentation transcript:

1 Radioimmunoassay, enzymoimmunoassay - principles, practical use.
Laboratory measurement of specific IgE antibodies. Laboratory measurement of autoantibodies. Flow cytometry - principles, practical use. Laboratory methods of assessment of cellular immunity. Laboratory methods of assessment of humoral immunity.

2 Humoral analytes Precipitation Aglutination Turbidimetry Nephelometry

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6 Imunofixation of serum

7 Humoral analytes Precipitation Aglutination Turbidimetry Nephelometry

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12 ELISA – assessment of specific antibodies

13 RAST

14 ELISA – assessment of antigen

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16 Indirect immunofluorescence

17 Asessment of cellular imunity
Number of cells - subpopulations Phagocytosis Activation of lymphocytes

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19 Flow cytometry

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21 Gating of lymphocytes

22 T and Th lymphocytes

23 T and B lymphocytes

24 Activated T lymphocytes

25 http://probes. invitrogen

26 IMPORTANT LEUKOCYTE POPULATIONS
CD3+ T lymphocytes: mature T lymphocytes % CD4+ T lymphocytes: helper T lymphocytes % CD8+ T lymphocytes: cytoxic T lymphocytes % CD25+ T lymphocytes: activated T lymphocytes % CD19+ B lymphocytes: mature B lymphocytes % CD56+ NK cells: natural killer cells % CD14+ cells: monocytes CD15+ cells: granulocytes CD38+ cells: plasma cells

27 TESTS OF LYMPHOCYTE FUNCTION
Ability to proliferate Ability to produce cytokines TESTS OF PHAGOCYTIC FUNCTION No of granulocytes Phagocytar activity of granulocytes Oxidative burst

28 Organ specific autoantibodies
Anti-thyroidal AB Anti-pancreatic beta cells AB Anti-acetylcholine receptor AB Anti-gonadal AB

29 Systemic autoantibodies
Anti-nuclear AB (ANA, ENA) Rheumatoid factor Anti-neutrophil cytoplasma AB (ANCA) Anti-phospholipid AB (APA)

30 Non-organ-specific autoantibodies in organ-localized autoimmune diseases
ANCA (non-specific inflammatory bowel disease) //+ASCA// ATG (coeliac disease) SS-A, SS-B (Sjögren syndrome)


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