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Mantoux – in vivo detection of specific cell immunity after exposition to antigen

Mantoux – in vivo detection of specific cell immunity after exposition to antigen - burden of patient by antigen - possible immunodeficiency of patients (anergy, risk of alergic reaction) - memory cell after BCG vaccination, - exposition to antigen = activation of MC

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Mantoux – in vivo detection of specific cell immunity after exposition to antigen

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  1. Mantoux – in vivo detection of specific cell immunity after exposition to antigen • - burden of patient by antigen • - possible immunodeficiency of patients (anergy, risk of alergic reaction) • - memory cell after BCG vaccination, • - exposition to antigen = activation of MC • - interpretation, • - booster dose of antigen,

  2. Matoux skin test reading • application • positive negative 15 mm

  3. Measurement of induration • 0 – 15 mm – negative, not enought, vaccination, revaccination • 15 mm – OK, positive (after vaccination ?) • 15 and more – too much, possible activity

  4. in vitro detection of specific cell immunity after exposition to the antigen • whole blood of patient to 3 vials covered with specific antigen (M. tbc) • specific memory cells after disease are activated antigen is not given to patient in vivo • no risk of exposition of immunocompromised patients

  5. Test based on exposition of whole blood in 3 vials • antigens of M.tbc, - 1 vial - non specific mitogen – positive control - without antigen – negative control

  6. Vials are incubated 16 – 24 hrs at 37 dg C (sensibilised memory cells are exposed to specific antigen and start production of INTERFERONU GAMA – cytokín of CMI Centrifugation Harvest the plasma Storage

  7. To detect IF-g secreted after in vitro activation (antigen on the surface of vial + memory cells in the whole blood of pacient) of memory cells produced after the infection (by M.tbc) we use ELISA test

  8. Interpretation: comparison of concentration of IFg in the vial with -TB ag : negat.contr. Negat control – exclude the nonspecific reaction Pozit.control – demonstrate capacity to react (negat. in imunocompromised or annergic

  9. Mantoux: in vivo reactivity of memory cells produced after vaccination (BCG) and after disease (M.tbc) application of i.d. memory cells migrate to the place of injection inflamation with induration Quantiferon: in vitro memory cells after M.tbc infection present in the blood in vial + M.tbc antigen = release of IFg ELISA for detection comparison

  10. detection of cytokines • In vitro ( immunity of patient is not influenced, boostered) • specificity • sensitivity • not cross reacting antibodies PERSPECTIVES • detection of latent infection (any infection if the antigen is available) – in practice now only M.tbc CMI - scientifical purposes

  11. Other use Detection of latent infection • CMV – in transplantation, • tissue parasitoses – withou serological reaction, • systemic mycoses, • detection of CMI or memory cells – for identification of memory after vaccination, when antibodies are no more present, • detection of non responders

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