Diagnostika alergie

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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.
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Humoral analytes

• Precipitation
• Aglutination
• Turbidimetry
• Nephelometry

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Imunofixation of serum
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Humoral analytes

• Precipitation
• Aglutination
• Turbidimetry
• Nephelometry

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ELISA assessment of specific antibodies

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RAST
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ELISA assessment of antigen
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Indirect immunofluorescence
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Asessment of cellular imunity

• Number of cells - subpopulations
• Phagocytosis
• Activation of lymphocytes

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Flow cytometry
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Gating of lymphocytes
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T and Th lymphocytes
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T and B lymphocytes
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Activated T lymphocytes
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http//probes.invitrogen.com/resources/education/t
utorials/4Intro_Flow/player.html
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IMPORTANT LEUKOCYTE POPULATIONS
CD3 T lymphocytes mature T lymphocytes
50 - 75 CD4 T lymphocytes helper
T lymphocytes 30 - 60 CD8 T lymphocytes
cytoxic T lymphocytes 15 - 30 CD25 T
lymphocytes activated T lymphocytes 1
- 5 CD19 B lymphocytes mature B
lymphocytes 5 - 15 CD56 NK cells
natural killer cells 5 - 15 CD14 cells
monocytes CD15 cells granulocytes CD38 cells
plasma cells
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TESTS OF LYMPHOCYTE FUNCTION
Ability to proliferate Ability to produce
cytokines
TESTS OF PHAGOCYTIC FUNCTION
No of granulocytes Phagocytar activity of
granulocytes Oxidative burst
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Organ specific autoantibodies

• Anti-thyroidal AB
• Anti-pancreatic beta cells AB
• Anti-acetylcholine receptor AB
• Anti-gonadal AB

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Systemic autoantibodies

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

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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)