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Advances in Flow Cytometry for Allergy Diagnosis

Learn about the rational application of flow cytometry in allergy diagnosis and monitoring. Explore various tests, including specific IgE, skin tests, and mediator release assays. Enhance your knowledge at our upcoming sessions in Barcelona.

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Advances in Flow Cytometry for Allergy Diagnosis

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  1. Rational application of flow cytometry in allergy Didier EBOImmunology – Allergology – RheumatologyUniversity of Antwerp - Belgium

  2. IgE-mediated allergy: diagnosis • History • Specific IgE (CAP)‏ • Skin test • Mediator release assays • histamine • leukotriene • Challenge tests All have draw backs None absolute Additional tests

  3. HLA-Dr / CD123/ CD63

  4. FCM in Barcelona • 4 sessions: • 9/6 Practical course: Cellular in vitro tests (119). • Principles and clinical applications • 10/6 Basic Plenary Symposium 3: Allergy diagnosis and monitoring (211/212)‏ • 10/6 In vitro allergy diagnosis (117). • Drug allergy • 10/6 (19h) EuroBAT Working group meeting (CCIB 124/125) • Drug, venom and food allergy • Over 30 abstracts

  5. Drug allergy • 15% ADR • > no IgE available • Skin tests • Culprit, cross-reactive compounds, alternatives • Serial dilutions (SPT, IDT)‏ • Provocation tests

  6. Stimulus Ref test Sens Spec N Ref -lactam H + ST 50 93 88 Sanz M, CEA 2002 -lactam H + ST + IgE + PT 49 91 110 Torres M, CEA 2004 -lactam 67 100 100 79 41 Abuaf N, CEA 2008 Metamizol H + PT 42 100 55 Gamboa P, Allergy 2003 NSAID H + PT 15-55 74-100 90 Gamboa P, CEA 2004 NMBA H 64 93 50 Abuaf N, JACI 1999 NMBA H + ST 54 100 60 Monneret G, Clin Imm 2002 NMBA H 79 100 24 Sudheer P, Anaesth 2005 NMBA H + ST 36-86 93 92 Kvedariene V, Allergy 2006 Rocuronium H + ST 92* 100 22 Ebo D, Allergy 2006 NSAID: non steroidal anti-inflammatory drugs NMBA: neuromuscular blocking agent * Including non-responders: 76%

  7. Rocuronium • NMBA • 2 NH4+ epitopes • Severe (life-threatening) anaphylaxis • Cross-reactivity • Diagnosis > skin tests • BAT • sIgE

  8. Group N History Skin test Controls* 21 + _ Allergics 24 + + NMBA controls* 18 + (other nmba)‏ _ * Other cause (e.g. NRL, dyes, chlorhexidine, …)‏ Rocuronium

  9. 17/21 (3 non-resp)‏ 81% 71% ROC 4% 21 24 18 rocu - rocu + nmba Adapted from: Ebo DG et al, Allergy 2006

  10. ROC 4% 21 24 18 rocu - rocu + nmba Adapted from: Ebo DG et al, Allergy 2006

  11. Ebo DG et al, Anaesth 2007

  12. * * neg. skin test rocuronium, pos skin test vecuronium

  13. Individual cases • Chlorhexidine • Rifamycin • Pentastarch, succinylated gelatin • Heparin and LMWH • Patent blue • Hyaluronidase • CsA (cremophore: polyethoxylated castor oil)‏ • Recombinant Hep B vaccine • Valcyclovir • Prednisolone

  14. Ebo DG et al, Allergy (in press)‏

  15. Ebo DG et al, Allergy 2005;60:703-704 Ebo DG et al, Allergy 2005

  16. Non-responders • Adults: 5-10% (patients, control individuals)‏ • Children: ? • Irrespective applied marker • CD63, CD203c, pp38 MAPK (preliminary data)‏ • Should be mentioned in publications !!! • Individual: test is lost • Studies: false-negatives

  17. Non-responder: refractory status • 4-6 weeks

  18. Non-responders: drugs • In vitro (supra therapeutical): yes • Desloratadin, promethazin • Atorvastatin, cervistatin • Majlesi Y, J Leuk Biol 2003 • In vitro (therapeutical): no • Desloratadin and glucocorticosteroids • Krauzelbinder et al. EAACI 2008 abstract 54.

  19. Gating the wrong cell See also Abuaf N, CEA 2008

  20. Expression of activation marker • CD63: bi-modal • CD203c, pp38 MAPK: homogeneous

  21. Ebo DG et al, (submitted)‏

  22. Drug allergy: • Diagnostic • Cross-reactivity • Alternatives • Combination of markers Complementary to other tests Absence of other tests Equivocal, false neg. results Less provocation tests

  23. Thanks

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