DOI QR코드

DOI QR Code

Basophil Markers for Identification and Activation in the Indirect Basophil Activation Test by Flow Cytometry for Diagnosis of Autoimmune Urticaria

  • Kim, Zehwan (Department of Clinical Pathology, Kyungpook National University School of Medicine) ;
  • Choi, Bong Seok (Department of Pediatrics, Kyungpook National University School of Medicine) ;
  • Kim, Jong Kun (Department of Emergency Medicine, Kyungpook National University School of Medicine) ;
  • Won, Dong Il (Department of Clinical Pathology, Kyungpook National University School of Medicine)
  • Received : 2015.02.16
  • Accepted : 2015.08.27
  • Published : 2016.01.01

Abstract

Background: The indirect basophil activation test using flow cytometry is a promising tool for autoimmune urticaria diagnosis. We aimed to identify better donor basophils (from atopic vs. non-atopic donors and interleukin-3 primed vs. unprimed basophils) and improve basophil identification and activation markers (eotaxin CC chemokine receptor-3 [CCR3] vs. CD123 and CD63 vs. CD203c). Methods: Donor basophils were obtained from non-atopic and atopic group O donors. Positive control sera were artificially prepared to simulate autoimmune urticaria patients' sera. Patient sera were obtained from nine children with chronic urticaria. Assay sensitivity was compared among each variation by using positive control sera (n=21), applying cutoff values defined from negative control sera (n=20). Results: For basophil identification, a combination of CCR3 and CD123 markers revealed a higher correlation with automated complete blood count (r=0.530) compared with that observed using CD123 (r=0.498) or CCR3 alone (r=0.195). Three activation markers on the atopic donor basophils attained 100% assay sensitivity: CD203c on unprimed basophils, CD63+CD203+ or CD63 alone on primed basophils; however, these markers on the non-atopic donor basophils attained lower assay sensitivity. Conclusions: For basophil identification markers, a combination of CD123 and CCR3 is recommended, while CD123 alone may be used as an alternative. Donor basophils should be obtained from an atopic donor. For basophil activation markers, either CD203c alone on unprimed basophils or CD203c and CD63 on primed basophils are recommended, while CD63 alone on primed basophils may be used as an alternative.

Keywords

References

  1. Marrouche N and Grattan C. Childhood urticaria. Curr Opin Allergy Clin Immunol 2012;12:485-90. https://doi.org/10.1097/ACI.0b013e3283574cb3
  2. Fiebiger E, Hammerschmid F, Stingl G, Maurer D. Anti-FcepsilonRIalpha autoantibodies in autoimmune-mediated disorders. Identification of a structure-function relationship. J Clin Invest 1998;101:243-51. https://doi.org/10.1172/JCI511
  3. Irinyi B, Szeles G, Gyimesi E, Tumpek J, Heredi E, Dimitrios G, et al. Clinical and laboratory examinations in the subgroups of chronic urticaria. Int Arch Allergy Immunol 2007;144:217-25. https://doi.org/10.1159/000103995
  4. Ebo DG, Bridts CH, Hagendorens MM, Aerts NE, De Clerck LS, Stevens WJ. Basophil activation test by flow cytometry: present and future applications in allergology. Cytometry B Clin Cytom 2008;74:201-10.
  5. Marone G, Spadaro G, Patella V, Genovese A. The clinical relevance of basophil releasability. J Allergy Clin Immunol 1994;94:1293-303. https://doi.org/10.1016/0091-6749(94)90345-X
  6. Zuberbier T, Schwarz S, Hartmann K, Pfrommer C, Czarnetzki BM. Histamine releasability of basophils and skin mast cells in chronic urticaria. Allergy 1996;51:24-8. https://doi.org/10.1111/j.1398-9995.1996.tb04545.x
  7. Szegedi A, Irinyi B, Gal M, Hunyadi J, Danko K, Kiss E, et al. Significant correlation between the CD63 assay and the histamine release assay in chronic urticaria. Br J Dermatol 2006;155:67-75. https://doi.org/10.1111/j.1365-2133.2006.07205.x
  8. Sabroe RA, Grattan CE, Francis DM, Barr RM, Kobza Black A, Greaves MW. The autologous serum skin test: a screening test for autoantibodies in chronic idiopathic urticaria. Br J Dermatol 1999;140:446-52. https://doi.org/10.1046/j.1365-2133.1999.02707.x
  9. Frezzolini A, Provini A, Teofoli P, Pomponi D, De Pita O. Serum-induced basophil CD63 expression by means of a tricolour flow cytometric method for the in vitro diagnosis of chronic urticaria. Allergy 2006;61: 1071-7. https://doi.org/10.1111/j.1398-9995.2006.01033.x
  10. Gyimesi E, Sipka S, Danko K, Kiss E, Hidvegi B, Gal M, et al. Basophil CD63 expression assay on highly sensitized atopic donor leucocytes-a useful method in chronic autoimmune urticaria. Br J Dermatol 2004; 151:388-96. https://doi.org/10.1111/j.1365-2133.2004.06042.x
  11. Ocmant A, Peignois Y, Mulier S, Hanssens L, Michils A, Schandene L. Flow cytometry for basophil activation markers: the measurement of CD203c up-regulation is as reliable as CD63 expression in the diagnosis of cat allergy. J Immunol Methods 2007;320:40-8. https://doi.org/10.1016/j.jim.2006.12.002
  12. Cozon G, Ferrandiz J, Peyramond D, Brunet J. Detection of activated basophils using flow cytometry for diagnosis in atopic patients. Allergol Immunopathol (Madr) 1999;27:182-7.
  13. Ebo DG, Lechkar B, Schuerwegh AJ, Bridts CH, De Clerck LS, Stevens WJ. Comments regarding 'Marked improvement of the basophil activation test by detecting CD203c instead of CD63' by Boumiza et al. Clin Exp Allergy 2003;33:849; author reply 852-3. https://doi.org/10.1046/j.1365-2222.2003.00691.x
  14. Gentinetta T, Pecaric-Petkovic T, Wan D, Falcone FH, Dahinden CA, Pichler WJ, et al. Individual IL-3 priming is crucial for consistent in vitro activation of donor basophils in patients with chronic urticaria. J Allergy Clin Immunol 2011;128:1227-34.e5. https://doi.org/10.1016/j.jaci.2011.07.021
  15. Marone G, Giugliano R, Lembo G, Ayala F. Human basophil releasability. II. Changes in basophil releasability in patients with atopic dermatitis. J Invest Dermatol 1986;87:19-23. https://doi.org/10.1111/1523-1747.ep12523520
  16. Sturm EM, Kranzelbinder B, Heinemann A, Groselj-Strele A, Aberer W, Sturm GJ. CD203c-based basophil activation test in allergy diagnosis: characteristics and differences to CD63 upregulation. Cytometry B Clin Cytom 2010;78:308-18.
  17. Wedi B, Novacovic V, Koerner M, Kapp A. Chronic urticaria serum induces histamine release, leukotriene production, and basophil CD63 surface expression--inhibitory effects of anti-inflammatory drugs. J Allergy Clin Immunol 2000;105:552-60. https://doi.org/10.1067/mai.2000.104939
  18. Chirumbolo S. The use of IL-3 in basophil activation tests is the real pitfall. Cytometry B Clin Cytom 2011;80:137-8; author reply 139.
  19. Hausmann OV, Gentinetta T, Fux M, Ducrest S, Pichler WJ, Dahinden CA. Robust expression of CCR3 as a single basophil selection marker in flow cytometry. Allergy 2011;66:85-91. https://doi.org/10.1111/j.1398-9995.2010.02431.x
  20. Ducrest S, Meier F, Tschopp C, Pavlovic R, Dahinden CA. Flowcytometric analysis of basophil counts in human blood and inaccuracy of hematology analyzers. Allergy 2005;60:1446-50. https://doi.org/10.1111/j.1398-9995.2005.00910.x
  21. Chirumbolo S, Ortolani R, Vella A. CCR3 as a single selection marker compared to CD123/HLADR to isolate basophils in flow cytometry: some comments. Cytometry A 2011;79:102-6.
  22. Ferrer M, Luquin E, Sanchez-Ibarrola A, Moreno C, Sanz ML, Kaplan AP. Secretion of cytokines, histamine and leukotrienes in chronic urticaria. Int Arch Allergy Immunol 2002;129:254-60. https://doi.org/10.1159/000066772
  23. Hennersdorf F, Florian S, Jakob A, Baumgartner K, Sonneck K, Nordheim A, et al. Identification of CD13, CD107a, and CD164 as novel basophil-activation markers and dissection of two response patterns in time kinetics of IgE-dependent upregulation. Cell Res 2005;15:325-35. https://doi.org/10.1038/sj.cr.7290301
  24. Yasnowsky KM, Dreskin SC, Efaw B, Schoen D, Vedanthan PK, Alam R, et al. Chronic urticaria sera increase basophil CD203c expression. J Allergy Clin Immunol 2006;117:1430-4. https://doi.org/10.1016/j.jaci.2006.02.016
  25. Laidlaw TM, Steinke JW, Tinana AM, Feng C, Xing W, Lam BK, et al. Characterization of a novel human mast cell line that responds to stem cell factor and expresses functional Fc${\epsilon}$RI. J Allergy Clin Immunol 2011; 127:815-22.e1-5. https://doi.org/10.1016/j.jaci.2010.12.1101
  26. Rashid A, Sadroddiny E, Ye HT, Vratimos A, Sabban S, Carey E, et al. Review: diagnostic and therapeutic applications of rat basophilic leukemia cells. Mol Immunol 2012;52:224-8. https://doi.org/10.1016/j.molimm.2012.05.019

Cited by

  1. Commentary: The Expression of CD123 Can Decrease with Basophil Activation: Implications for the Gating Strategy of the Basophil Activation Test vol.7, pp.None, 2016, https://doi.org/10.3389/fimmu.2016.00260
  2. Reply to the Letter to the Editor by Dr. Chirumbolo vol.36, pp.5, 2016, https://doi.org/10.3343/alm.2016.36.5.498
  3. Basophil Activation Test for Chronic Urticaria vol.36, pp.5, 2016, https://doi.org/10.3343/alm.2016.36.5.499
  4. Neutrophil oxidative burst as a diagnostic indicator of IgG-mediated anaphylaxis vol.53, pp.4, 2016, https://doi.org/10.5045/br.2018.53.4.299
  5. Prueba de activación de basófilos: aspectos técnicos, metodológicos y su utilidad clínica vol.66, pp.3, 2016, https://doi.org/10.15446/revfacmed.v66n3.61820
  6. EAACI/GA2LEN/EDF/WAO-Leitlinie für die Definition, Klassifikation, Diagnose und das Management der Urtikaria - konsentierte, deutschsprachige Übersetzung : Leitlinie der European Academy of vol.27, pp.8, 2018, https://doi.org/10.1007/s15007-018-1751-3
  7. Urticaria crónica en niños. Revisión sistemática vol.29, pp.1, 2016, https://doi.org/10.35366/93322
  8. How I investigate basophilia in daily practice vol.42, pp.3, 2016, https://doi.org/10.1111/ijlh.13146
  9. Basophil counting in hematology analyzers: time to discontinue? vol.59, pp.5, 2021, https://doi.org/10.1515/cclm-2020-1528
  10. Basophil activation: theoretical aspects and use in the diagnosis of allergic diseases vol.23, pp.3, 2016, https://doi.org/10.15789/1563-0625-bat-2174
  11. Validation of inducible basophil biomarkers: Time, temperature and transportation vol.100, pp.6, 2016, https://doi.org/10.1002/cyto.b.21991
  12. Profiling Glioblastoma Cases with an Expression of DCX, OLIG2 and NES vol.22, pp.24, 2021, https://doi.org/10.3390/ijms222413217