Original Articles
Vol. 17 No. 1 (2025): Review Articles, Original Article, Scientific Letter, Case Reports Letter to the Editor
INTEGRATION OF CD200, CD43 AND ROR1 IN MULTIPARAMETER FLOW CYTOMETRY (MFC) ROUTINE PANELS FOR THE DIFFERENTIAL DIAGNOSIS OF B-CELL LYMPHOPROLIFERATIVE DISORDERS (B-LPDS)
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All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Received: October 1, 2024
Accepted: November 20, 2024
Accepted: November 20, 2024
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Background: Clonal mature B-cell lymphoproliferative disorders (B-LPDs) are a heterogeneous group of neoplasia characterized by the proliferation of mature B lymphocytes in the peripheral blood, bone marrow and/or lymphoid tissues. B-LPDs classification into different subtypes and their diagnosis are based on a multiparametric approach, however accurate diagnosis may be challenging, especially for cases of ambiguous interpretation. Multiparameter flow cytometry (MFC) represents an extensively used technique to detect the presence of different cellular lines in immunology and hematology. MFC results provide an essential contribution to B-LPDs diagnostic process, even more considering that panels are constantly integrating novel markers to improve diagnostic accuracy.
Objectives: The aim was to evaluate the contributing role of MFC routinary studies analyzing the expression and the mean fluorescence intensity (MFI) of CD200, ROR1 and CD43 in various B-LPDs, to evaluate their usefulness in differential diagnosis of these diseases.
Methods: We retrospectively evaluated a total of 2615 consecutive cases of newly samples (collected mostly from patients with lymphocytosis) analyzed by MFC carried out in B-LPDs diagnostic process referred to the Division of Hematology of ‘Sapienza’ University of Rome. We compared results of CD200, ROR1 and CD43 expression percentage and their MFI between different subtypes of B-LPDs.
Results: In chronic lymphocytic leukemia (CLL) CD200, ROR1 and CD43 were always expressed with bright intensity. CLL samples presented CD200 expression and MFI [CD200%, mean: 100 (range, 24-100); positivity rate: 100%; MFI, mean = 168 (range, 10-1200)] and highly expressed statistically higher than mantle cell lymphoma (MCL) (p<0.001), which is usually negative for CD200, and variant hairy cell leukemia (vHCL, according to 2022 ICC) (p<0.001), but comparable with classic HCL (cHCL) (p>0.9). ROR1 resulted expressed in all CLL [ROR1%, mean: 100 (range, 52-100), positivity rate: 100%; MFI, mean 53 (range, 10-202)] and MCL cases with comparable MFI (p>0.9). CD43 expression and MFI were significantly higher in CLL [CD43%, mean: 99 (range, 59-100); positivity rate: 100%; MFI, mean = 218 (range, 41-980)] than in MCL, vHCL, cHCL and all the others mature B-cell neoplasia (p<0.001). CD200 and CD43 expression and MFI were significantly higher in cHCL compared to vHCL. Among the others mature B-cell neoplasia, CD200 was variably expressed in follicular lymphoma (FL), marginal zone lymphoma (MZL), diffuse large B-cell lymphoma (DLBCL) and lymphoplasmacytic lymphoma (LPL). ROR1 and CD43 presented a very low expression percentage in this latter group, being mostly negative. Persistent polyclonal B-cell lymphocytosis (PPBL) resulted uniformly positive for CD200 and negative for ROR1 and CD43.
Conclusions: Our data suggest that evaluation of CD200, ROR1 and CD43 antigens and their intensity of expression, along with commonly used markers in MFC routine panels for B-LPDs, might be extremely useful for a prompt diagnostic evaluation in the differential diagnosis of these diseases.
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Citations
Al-Zubaidi HK, Hughes SF. The use of CD200 in the differential Diagnosis of B-Cell lymphoproliferative disorders. Br J Biomed Sci.2023;80:11573.
WHO Classification of Tumours Editorial Board. Haematolymphoid tumours. Lyon (France): International Agency for Research on Cancer; 2024. (WHO classification of tumours series, 5th ed.; vol. 11). https://publications.iarc.who.int/637
Bisegna ML, Cordone I, Peragine N, Milani ML, Intoppa S, de Fabritiis P, Martelli M, De Propris MS. Neoplastic bone marrow invasion:rapid exclusion of hematological disease by flow cytometric routine panels. Blood Cells Mol Dis. 2023 Mar;99:102721
De Propris MS, Musiu P, Intoppa S, Nardacci MG, Pucciarini A, Santi A, Peragine N, Canichella M, De Luca ML, D'Elia GM, Del Giudice I, Pulsoni A, Falini B, Guarini A, Martelli M, Tiacci E, Foà R. Hairy cell leukaemia with low CD103 expression: A rare but important diagnostic pitfall. Br J Haematol. 2022 Jul;198(2):e28-e31.
Braylan RC. Impact of flow cytometry on the diagnosis and characterization of lymphomas, chronic lymphoproliferative disorders and plasma cell neoplasia. Cytometry. 2004; 58(1):57-61.
De Propris MS, Intoppa S, Milani ML, Mariglia P, Nardacci MG, Peragine N, Foà R, Guarini A. ROR1 is an accurate and reliable marker of minimal residual disease in chronic lymphocytic leukaemia. Br J Haematol. 2020 Sep;190(6):e346-e349.
El Desoukey NA, Afify RA, Amin DG, Mohammed RF. CD200 expression in B-cell chronic lymphoproliferative disorders. J Investig Med. 2012;60(1):56-61.
De Propris MS, Raponi S, Diverio D, Milani ML, Meloni G, Falini B, Foà R, Guarini A. High CD33 expression levels in acute myeloid leukemia cells carrying the nucleophosmin (NPM1) mutation. Haematologica. 2011 Oct;96(10):1548-51. doi: 10.3324/haematol.2011.043786. Epub 2011 Jul 26.
Raponi S, De Propris MS, Intoppa S, Milani ML, Vitale A, Elia L, Perbellini O, Pizzolo G, Foá R, Guarini A. Flow cytometric study of potential target antigens (CD19, CD20, CD22, CD33) for antibody-based immunotherapy in acute lymphoblastic leukemia: analysis of 552 cases. Leuk Lymphoma. 2011 Jun;52(6):1098-107.
Arlindo EM, Marcondes NA, Fernandes FB, Faulhaber GAM. Quantitative flow cytometric evaluation of CD200, CD123, CD43 and CD52 as a tool for the differential diagnosis of mature B-cell neoplasms. Rev Bras Hematol Hemoter. 2017;39(3):252-258.
Kroft SH, Harrington AM. Flow Cytometry of B-Cell Neoplasms. Clin Lab Med. 2017 Dec;37(4):697-723. doi: 10.1016/j.cll.2017.07.001. Epub 2017 Aug 31.
Moreau EJ, Matutes E, A'Hern, RP, Morilla AM, Morilla RM, Owusu-Ankomah KA., Seon, BK, Catovsky, D. Improvement of the chronic lymphocytic leukemia scoring system with the monoclonal antibody SN8 (CD79b). Am J Clinical Pathol.1997;108(4):378–382.
Rawstron AC, Böttcher S, Letestu R, Villamor N, Fazi C, Kartsios H, de Tute RM, Shingles J, Ritgen M, Moreno C, Lin K, Pettitt AR, Kneba M, Montserrat E, Cymbalista F, Hallek M, Hillmen P, Ghia P; European Research Initiative in CLL. Improving efficiency and sensitivity: European Research Initiative in CLL (ERIC) update on the international harmonised approach for flow cytometric residual disease monitoring in CLL. Leukemia. 2013 Jan;27(1):142-9.
Rawstron AC, Kreuzer K-A, Soosapilla A, Spacek M, Stehlikova O, Gambell P, McIverBrown N, Villamor N, Psarra K, Arroz M, Milani R, de la Serna J, Cedena MT, Jaksic O, Nomdedeu J, Moreno C, Rigolin GM, Cuneo A, Johansen P, Johnsen HE, Rosenquist R, Utoft Niemann C, Kern W, Westerman D, Trneny M, Mulligan S, Doubek M, Pospisilova S, Hillmen P, Oscier D, Hallek M, Ghia P, Montserrat E. Reproducible diagnosis of chronic lymphocytic leukemia by flow cytometry: An european research initiative on CLL (ERIC) & European Society for Clinical Cell Analysis (ESCCA) Harmonisation Project. Cytometry B Clinic Cytom. 2018; 94(1): 121–128.
McCaughan GW, Clark MJ, Barclay AN. Characterization of the Human Homolog of the Rat MRC OX-2 Membrane Glycoprotein. Immunogenetics.1987;25(5):329–35.
Wright GJ, Jones M, Puklavec MJ, Brown MH, Barclay AN. The unusual distribution of the neuronal/lymphoid cell surface Cd200 (Ox2) glycoprotein is conserved in humans. Immunology. 2001;102(2):173–9.
Nathan C, Muller WA. Putting the brakes on innate immunity: A regulatory role for CD200? Nat Immunol. 2001;2(1):17–9.
Palumbo GA, Parrinello N, Fargione G, Cardillo K, Chiarenza A, Berretta S, Conticello C, Villari L, Di Raimondo F. CD200 expression may help in differential diagnosis between mantle cell lymphoma and B-cell chronic lymphocytic leukemia. Leuk Res. 2009 Sep;33(9):1212-6.
Green JL, Kuntz SG, Sternberg PW. Ror receptor tyrosine kinases: orphans no more. Trends Cell Biol 2008;18(11):536–44.
Daneshmanesh AH, Mikaelsson E, Jeddi-Tehrani M, Bayat AA, Ghods R, Ostadkarampour M, Akhondi M, Lagercrantz S, Larsson C, Osterborg A, Shokri F, Mellstedt H, Rabbani H. Ror1, a cell surface receptor tyrosine kinase is expressed in chronic lymphocytic leukemia and may serve as a putative target for therapy. Int J Cancer 2008;123(5):1190–5.
Yoda A, Oishi I, Minami Y. Expression and function of the Ror-family receptor tyrosine kinases during development: lessons from genetic analyses of nematodes, mice, and humans. J Recept Signal Transduct Res 2003;23(1):1–15.
Baskar S, Kwong KY, Hofer T, Levy JM, Kennedy MG, Lee E, Staudt LM, Wilson WH, Wiestner A, Rader C. Unique cell surface expression of receptor tyrosine kinase ROR1 in human B-cell chronic lymphocytic leukemia. Clin Cancer Res 2008;14(2):396–404.
Barna G, Mihalik R, Timár B, Tömböl J, Csende Z, Sebestyén A, Bödör C, Csernus B, Reiniger L, Peták I, Matolcsy A. ROR1 expression is not a unique marker of CLL. Hematol Oncol. 2011;29(1):17-21.
Rawstron AC, de Tute RM, Shingles J, Gorman L, Turner K, Evans PAS, Barrans SL, O’Connor SJM, Burton C, Owen RG, Hillmen P. Improving the Differential Diagnosis of CD51 B-Lymphoproliferative Disorders. http://learningcenter.ehaweb.org/eha/2016/21st/133474/andy.rawstron.improving.the.differential.diagnosis.of.cd52B.html?f5m1
Rosenstein Y, Santana A, Pedraza-Alva G. CD43, a molecule with multiple functions.
Immunol Res. 1999;20(3):89–99.
Sorigue M, Juncà J, Sarrate E, Grau J. Expression of CD43 in chronic lymphoproliferative leukemias. Cytometry B Clin Cytom. 2018;94(1):136-142.
Pedreira CE, Costa ES, Almeida J, Fernandez C, Quijano S, Flores J, Barrena S, Lecrevisse Q, Van Dongen JJ, Orfao A; EuroFlow Consortium. A probabilistic approach for the evaluation of minimal residual disease by multiparameter flow cytometry in leukemic B-cell chronic lymphoproliferative disorders. Cytometry A. 2008 Dec;73A(12):1141-50.
Van Dongen JJ, Lhermitte L, Böttcher S, Almeida J, van der Velden VH, Flores-Montero J, Rawstron A, Asnafi V, Lécrevisse Q, Lucio P, Mejstrikova E, Szczepański T, Kalina T, de Tute R, Brüggemann M, Sedek L, Cullen M, Langerak AW, Mendonça A, Macintyre E, Martin-Ayuso M, Hrusak O, Vidriales MB, Orfao A; EuroFlow Consortium (EU-FP6, LSHB-CT-2006-018708). EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia. 2012 Sep;26(9):1908-75.
Campo E, Swerdlow SH, Harris NL, Pileri S, Stein H, Jaffe ES. The 2008 WHO classification of lymphoid neoplasms and beyond: evolving concepts and practical applications. Blood. 2011 May 12;117(19):5019-32.
Swerdlow SH, Campo E, Pileri SA, Harris NL, Stein H, Siebert R, Advani R, Ghielmini M, Salles GA, Zelenetz AD, Jaffe ES. The 2016 revision of the World Health Organization classification of lymphoid neoplasms. Blood. 2016 May 19;127(20):2375-90.
Grever M, Andritsos L, Anghelina M, Arons E, Banerji V, Barrientos J, Bhat SA, Blachly J, Broccoli A, Call T, Dearden C, Dietrich S, Else M, Epperla N, Fagarasanu A, Falini B, Forconi F, Gozzetti A, Hampel P, Hermel DJ, Iyengar S, Johnston JB, Juliusson G, Kreitman RJ, Lauria F, Lozanski G, Oakes CC, Parikh SA, Park J, Quest G, Rai K, Ravandi F, Robak T, Rogers KA, Saven A, Seymour JF, Tadmor T, Tallman MS, Tam CS, Tiacci E, Troussard X, Wörmann B, Zent CS, Zenz T, Zinzani PL. Hairy cell leukemia variant and WHO classification correspondence Re: 5th edition WHO classification haematolymphoid tumors: lymphoid neoplasms. Leukemia. 2024 Jul;38(7):1642-1644. doi: 10.1038/s41375-024-02280-0. Epub 2024 Jun 8.
Campo E, Jaffe ES, Cook JR, Quintanilla-Martinez L, Swerdlow SH, Anderson KC, Brousset P, Cerroni L, de Leval L, Dirnhofer S, Dogan A, Feldman AL, Fend F, Friedberg JW, Gaulard P, Ghia P, Horwitz SM, King RL, Salles G, San-Miguel J, Seymour JF, Treon SP, Vose JM, Zucca E, Advani R, Ansell S, Au WY, Barrionuevo C, Bergsagel L, Chan WC, Cohen JI, d'Amore F, Davies A, Falini B, Ghobrial IM, Goodlad JR, Gribben JG, Hsi ED, Kahl BS, Kim WS, Kumar S, LaCasce AS, Laurent C, Lenz G, Leonard JP, Link MP, Lopez-Guillermo A, Mateos MV, Macintyre E, Melnick AM, Morschhauser F, Nakamura S, Narbaitz M, Pavlovsky A, Pileri SA, Piris M, Pro B, Rajkumar V, Rosen ST, Sander B, Sehn L, Shipp MA, Smith SM, Staudt LM, Thieblemont C, Tousseyn T, Wilson WH, Yoshino T, Zinzani PL, Dreyling M, Scott DW, Winter JN, Zelenetz AD. The International Consensus Classification of Mature Lymphoid Neoplasms: a report from the Clinical Advisory Committee. Blood. 2022 Sep 15;140(11):1229-1253. doi: 10.1182/blood.2022015851. Erratum in: Blood. 2023 Jan 26;141(4):437.
Del Giudice I, Pileri SA, Rossi M, Sabattini E, Campidelli C, Starza ID, De Propris MS, Mancini F, Perrone MP, Gesuiti P, Armiento D, Quattrocchi L, Tafuri A, Amendola A, Mauro FR, Guarini A, Foà R. Histopathological and molecular features of persistent polyclonal B-cell lymphocytosis (PPBL) with progressive splenomegaly. Br J Haematol. 2009 Mar;144(5):726-31.
Sandes AF, Chauffaille M de L, M.C. Oliveira CR, Maekawa Y. Tamashiro N, Takao TT, Ritter EC, Rizzatti EG. CD200 Has an important role in the differential diagnosis of mature b-cell Neoplasms by multiparameter flow cytometry. Cytometry B Clin Cytom. 2014; 86B: 98–105.
Palumbo GA, Parrinello N, Fargione G, Cardillo K, Chiarenza A, Berretta S, Conticello C, Villari L, Di Raimondo F. CD200 expression may help in differential diagnosis between mantle cell lymphoma and B-cell chronic lymphocytic leukemia. Leuk Res. 2009; 33(9):1212-6. Epub 2009 Feb 20.
Alapat D, Coviello-Malle J, Owens R, Qu P, Barlogie B, Shaughnessy JD, Lorsbach RB. Diagnostic Usefulness and Prognostic Impact of CD200 Expression in Lymphoid Malignancies and Plasma Cell Myeloma. Am J Clin Pathol. 2012; 137(1): 93–100.
El Desoukey NA, Dalia Gamil Amin, Aleem Afify RA, Mohammed RA. CD200 expression in B-cell chronic lymphoproliferative disorders. J Investig Med. 2012;60(1):56-61.
Pillai V, Pozdnyakova O, Charest K, Li B, Shahsafaei A, Dorfman DM. CD200 flow cytometric assessment and semiquantitative immunohistochemical staining distinguishes hairy cell leukemia from hairy cell leukemia-variant and other B-cell lymphoproliferative disorders. Am J Clin Pathol. 2013;140(4), 536–543.
Salem DA, Scott D, McCoy CS, Liewehr DJ, Venzon DJ, Arons E, Kreitman RJ, Stetler-Stevenson M, Yuan CM. Differential expression of CD43, CD81, and CD200 in classic versus variant hairy cell leukemia. Cytometry B Clin Cytom. 2019; 96B: 275–282. Epub 2019 May 11.
Uhrmacher S, Schmidt C, Erdfelder F, Poll-Wolbeck SJ, Gehrke I, Hallek, M, Kreuzer, K. A. Use of the receptor tyrosine kinase-like orphan receptor 1 (ROR1) as a diagnostic tool in chronic lymphocytic leukemia (CLL). Leuk Res. 2011;35(10):1360-6. Epub 2011 Apr 30.
De Propris MS, Intoppa S, Milani ML, Mariglia P, Nardacci MG, Peragine N, Foà R, Guarini A. ROR1 is an accurate and reliable marker of minimal residual disease in chronic lymphocytic leukaemia. Br J Haematol. 2020;190(6):e346-e349. Epub 2020 Jun 24.
Rawstron AC, Villamor N, Ritgen M, Böttcher S, Ghia P, Zehnder JL, Lozanski G, Colomer D, Moreno C, Geuna M, Evans PA, Natkunam Y, Coutre SE, Avery ED, Rassenti LZ, Kipps TJ, Caligaris-Cappio F, Kneba M, Byrd JC, Hallek MJ, Montserrat E, Hillmen P. International standardized approach for flow cytometric residual disease monitoring in chronic lymphocytic leukaemia. Leukemia. 2007 May;21(5):956-64.
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B-Cell Lymphoproliferative disorders (B-LPDs), Multiparameter flow cytometry (MFC)Supporting Agencies
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“INTEGRATION OF CD200, CD43 AND ROR1 IN MULTIPARAMETER FLOW CYTOMETRY (MFC) ROUTINE PANELS FOR THE DIFFERENTIAL DIAGNOSIS OF B-CELL LYMPHOPROLIFERATIVE DISORDERS (B-LPDS)” (2024) Mediterranean Journal of Hematology and Infectious Diseases, 17(1), p. e2025.002. doi:10.4084/MJHID.2025.002.
Copyright (c) 2024 Alessandro Laganà, Raffaele Maglione, Alessandro Costa, Biancamaria Mandelli, Maria Laura Bisegna, Maria Laura Milani, Valeria Filipponi, Maria Grazia Nardacci, Tania Soriano, Eugenio Santacroce, Luigi Petrucci, Carla Giordano, Maurizio Martelli, Maria Stefania De Propris
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