Research Articles







AL Marshad Ibrahim, Hassanein Nagwa, Balakrishnan Bargavi

ABSTRACT: Although the phenotypic features of basophils have been highlighted in literatures, yet some labs generate confusion while discriminating blasts from basophil gate in flow cytometry analysis. This is sometimes due to the overlap of different population on CD45 side scatter plot, and in other situations due to lack of markers which highlight basophils due to financial jeopardy. Aim of the study: Selection of a proper combination of markers with unique expression on basophils to clearly highlight basophils especially in under-resourced labs with limited budget. Methods: Comprehensive basophil phenotyping  applied on 40newly diagnosed cases of CML and 20 cases of regenerating marrow post chemotherapy for Precursor B lymphoblastic leukemia with no evidence of abnormal cellsas control.  Flow cytometry markers used were CD45, CD38, CD71, CD33, CD22, CD123, HLA-DR, CD34, CD117, CD3, CD4, CD8, CD56, CD11b, HLA-DR and CD15. Results: Basophils from patients and control samples were clearly discriminated from blasts and lymphocytes by using CD123 and CD22, a unique pattern of expression with perfect back-gating on CD45 side scatter plot which proved the discrimination. Conclusion: CD123 and CD22 will clearly discriminate basophils from different types of population which may occupy the window area as myeloblast, precursor B cells (hematogones), abnormal B cells (lymphoblasts) and mature B cells. This combination should therefore be part of the screening panel used as first line along with CD45in labs with limited budget.


KEY WORDS: Basophil, CML, CD123, CD22.


  1. MC, Souvignet A, Pont J, Solly F, Mondet J, KesrS, Pernollet M, DumestrePerard C, CamposL, Cesbron JY. One tube with eight antibdies for 14-part bone marrow leukocyte differential using flowcytometry. Cytometry B Clin Cytom 2017 Jul; 92(4): 299-309. doi: 10.1002/cyto.b.21369. Epub 2016 May 6.S

  2. Proven D,Singel C,Baglin T, Dokal I,2015. White blood cells Oxford Handbook of clinical hematology. Fourth edition, chapter 13, Oxyford University Press, Great clarend on street, oxford,OX26DP,UK,page 116.

  3. Han X, Jorgensen JL, Brahmandam A, Schlette E, Huh YO, Shi Y, Awagu S, Chen W.   of basophils by  . Arch Pathol Lab Med. 2008 May; 132(5):813-9. doi: 10.1043/1543-2165(2008)132[813:ISOBBM]2.0.CO;2.

  4. Toba K, Hanawa H, Fuse I, Sakaue M, Watanabe K, Uesugi Y, Higuchi W, Takahashi M, Aizawa Y. Difference in CD22 molecules in human B cells and basophils .Experimental Hematology , 2002 ,Volume 30 , Issue 3 , 205 – 21.

  5. Hassanein NM, Alcancia F, Perkinson KR, Buckley PJ, LagooAS. Distinct expression patterns of CD123 and CD34 on normal bone marrow B-cell precursors ("hematogones") and B lymphoblastic leukemia blasts. Am J ClinPathol. 2009 Oct;132(4):573-80. doi: 10.1309/AJCPO4DS0GTLSOEI.

  6. Santos AF, Bécares N, Stephens A, Turcanu V, Lack G. The  of  can  with  : implications for the strategy of the   . Clin Transl Allergy. 2016 Apr 1;6:11. doi: 10.1186/s13601-016-0100-4. E Collection 2016.

  7. Gönen, M., Sun, Z., Figueroa, M. E., Patel, J. P., Abdel-Wahab, O., Racevskis, J., Ketterling, R. P., Fernandez, H., Rowe, J. M., Tallman, M. S., Melnick, A., Levine, R. L. Paietta, E. CD25 expression status improves prognostic risk classification in AML independent of established biomarkers: ECOG phase 3 trial, E1900. Blood,(2012). 120(11), 2297-306.

  8. Agis, H., Füreder, W., Bankl, H. C., Kundi, M., Sperr, W. R., Willheim, M., Boltz-Nitulescu, G., Butterfield, J. H., Kishi, K., Lechner, K., …Valent, P. (Comparative immunophenotypic analysis of human mast cells, blood basophils and monocytes. 1996)., (4), 535-43.

  9. Saksena A, Gautam P, and Singh TSide scatter versus CD45 flow cytometric plot can distinguish acute leukaemia subtypes. Indian J Med Res. 2016 May; 143(Suppl 1): S17–S22..

  10. Gajendra S, flow cytometery in acute leukemia. Clinics in oncology,2016,volume1,article1166.

  11. Sidhwa K, Mansukhani D, Padate B, Gupta S and Khodaiji S. The Basophil Dilemma. Ann HematolOncol. 2017; 4(2): 1137. ISSN:2375-7965

  12. Orfao, A., Escribano, L., Villarrubia, J., Velasco, J. L., Cerveró, C., Ciudad, J., Navarro, J. L., … San Miguel, J. F. (1996). Flow cytometric analysis of mast cells from normal and pathological human bone marrow samples: identification and enumeration. , (5), 1493-9.

  13. ChirumboloS ,Bjørklund G , Vella A, Using a CD45dim/CD123bright/HLA-DRnegphenotyping protocol to gate basophils in FC for airway allergy. CD123 does not decrease. Adv. Respir. Med. 2017; 85: 193–201.

  14. ÇehreliC.Diagnostic Problems in Chronic Basophilic Leukemia Turk J Haematol. 2018 Nov 13;35(4):283-289. doi: 10.4274/tjh.2018.0129. Epub 2018 Aug 1. Review.

  15. Stacchini A, Demurtas A, GodioL.   detection of degranulated basophils in chronic  leukemia in accelerated phaseCytometry B Clin Cyto. 2011 Mar; 80(2):122-4. doi: 10.1002/cyto.b.20566. Epub 2010 Sep 15.

  16. Orfao, A., Escribano, L., Villarrubia, J., Velasco, J. L., Cerveró, C., Ciudad, J., Navarro, J. L., … San Miguel, J. F. (1996). Flow cytometric analysis of mast cells from normal and pathological human bone marrow samples: identification and enumeration. , (5), 1493-9.