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Immunology iv.serology
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Immunology iv.serology

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  • 1. Serological reactions
  • 2. (Polyclonal) antisera
    • Obtained a from animals (rabbits, goats, horses) after repeated immunisation by antigen .
    • Markedly polyreactive – antibodies bind to many epitopes of the antigen but also with other antigens .
    • This is advantageous in „classical“ serological reactions (agglutination, precipitation) .
  • 3. Clonal selection theory
  • 4. Myeloma
    • Tumor derived from plasma cell
    • The tumor cells retain the capacity to secrete immunoglobulins
    • The secreted immunoglobu li n is a paraprotein - all secreted molecules have the same variable region (= re a ct wit h only one concrete epitope)
  • 5. Monoclonal antibodies
    • Prepared by immortali z ation of B-cells from an i mmun i zed mouse .
    • Hybrido m a is composed of a n antigen-specific B- cell and mouse myeloma cell .
    • Pr oduced antibodies are stric t ly monospecific and therefore cannot be used in several „classical“ serological reactions (agglutination, precipitation) .
  • 6.  
  • 7. Laboratory use of monoclonal antibodies
    • Highly specific agent used for ELISAs, RIAs, determination of cells su r face antigens…
    • Because the y react onl y with a si ngle epitope, nu m ber of „bridges“ is to low to overcome repulsive forces in classical reacti o ns like agglutination or precipitation.
  • 8. Clinical use of monoclonal antibodies
    • Immunosuppressive treatment (anti CD3, CD54, CD20)
    • Antinflammatory treatment –
      • Cytokine neuralisation (anti- TNF  )
      • Adhesion molecules blocade (anti-LFA-1….)
    • Anti-tumor treatment (anti-CD20…)
    • Anti aggregation treatment (anti- gpIIb-IIIa)
    • Antitoxins (digoxin)
  • 9. Two phases of serological reaction
    • Primary phase – concrete antibody (with its variable region must be present) binds to a concrete epitope. = Specific phase of the reaction
    • Secondary phase – vizualization of the fact of previously occurred primary reaction.
  • 10. Serological reactions
    • Agglutinatin
    • Precipitation
    • Immunoassays
      • RIA
      • ELISA
      • Immunofluorescence
    • Reactions based on activation of complement cascad e by complex-antigen-antibody
    • Reactions based on neutral is ation of some biologic effect of antigen
  • 11. Agglutination
    • Reaction between antiserum and corpuscular antigen (erythrocyte, bacterium, latex corpuscle). The corpuscles are clumped together, which morfologically expressed as agglutinate.
  • 12.  
  • 13.  
  • 14.
    • Complete antibodies : after reaction with antigen cause visible agglutionation or precipitation reaction
    • Incomplete antibodies : despite the fact that the reaction between epitope and antibody occurred, the agglutinate or precipitate cannot be detected.
    • Cause: movalent antibody (IgA), low number of bridges between antigens., to intense repulsive forces between antigens…
  • 15. Coombs test
  • 16.  
  • 17. Precipitation
    • Reaction between polyclonal antiserum and soluble (molecular) antigen . A complex lattice of interlocking aggregates is formed. If performed in a soluti o n the precipitate falls out of the solution.
  • 18. Immunodiffusion-I
  • 19. Immunodiffusion - II
  • 20. Immunodiffusion - III
  • 21.  
  • 22. ELISA
  • 23. ELISA
  • 24.  
  • 25.  
  • 26. ELISA READER
  • 27. Imunofluorescence - direct - indirect
  • 28.
    • Přímá
  • 29.
    • Nepřímá
    Sérum
  • 30. ANA Pozitive granular type
  • 31. ANA – homoge nous type