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,

Matoux skin test reading

application positive negative

15 mm

Measurement of induration 0 – 15 mm – negative, not enought,

vaccination, revaccination 15 mm – OK, positive (after vaccination ?) 15 and more – too much, possible activity

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

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

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

To detect IF- 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

Interpretation:comparison of concentration of IF 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

comparison 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 IF

ELISA for detection

detection of cytokines In vitro ( immunity of patient is not influenced, boostered) specificity sensitivity not cross reacting antibodiesPERSPECTIVES detection of latent infection (any

infection if the antigen is available) – in practice now only M.tbc CMI

- scientifical purposes

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