3rd International ELN Workshop:
Standardization of Flow Cytometry in MDS
London, Greenwich 4-6 November 2010
a.a. van de loosdrecht, chair
r. ireland, co-chair
update ELNet wp8; ASH Orlando, December 4, 2010
and ELNet WP conference, Mannheim, February 1, 2011
VU University Medical Center
Amsterdam The Netherlands
3rd International ELN Workshop:
Standardization of Flow Cytometry in MDS
London, Greenwich 4-6 November 2010
a.a. van de loosdrecht, chair
r. ireland, co-chair
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12 countries; 28 participants; in- and outside Europe
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Austria: P. Valent
France: MC Bene, J. Feuillard
Germany: W. Kern
Great Britain: R. Ireland, U. Johansson, M. Cullen, G. Mufti, T. Minle, S. Kordasti
Italy: M. della Porta [absent]
Netherlands: C. Alhan, A.A. van de Loosdrecht, J te Marvelde, V. van de
Velden, TM Westers, B. Moshaver, T deWitte
Spain: [absent]
Sweden: A. Porwit
Greece: K. Psarra
Japan: K. Ogata
USA: D. Wells, M. Loken, J Cutler
Australia: K. Burbury
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VU University Medical Center
Amsterdam The Netherlands
Goal:
• define minimal flow cytometric criteria to analyse MDS
•  ELN 2011 [2 consensus documents: rationale/technical]
• Define: role of FCM:
– Diagnosis
– Prognosis
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predictive value for R/ [emerging new drugs]
disease monitoring
VU University Medical Center
Amsterdam The Netherlands
Proposed antigens of importance in FCM in MDS:
ELN consensus 2009
ELN consensus document MDS and flow cytometry:
van de Loosdrecht, ELN members WP8, et al., Haematologica 2009;94:1124-1134
VU University Medical Center
Amsterdam The Netherlands
BONE MARROW SUB SET
RECOMMENDED ANALYSIS
Overview of some recommended flow cytometric analyses of
subsets in the BM by the ELNet; London 2010
Erythroid compartment
% of nucleated erythroid cells
Relation CD71 and CD235a
Expression of CD71
Immature myeloid and monocytic progenitors
% of cells
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+
CD34 CD45 in combination with FCS/SSC;
expression of CD13 and CD117;
expression of HLA-DR
expression of CD5, CD7, CD56;
asynchronous expression of CD11b, CD15
Granulocytes
% of cells
SSC as ratio vs. SSC of lymphocytes
relation of CD13 and CD11b
relation of CD13 and CD16
expression of CD10 on mature neutrophils ??
Monocytes
% of cells
relation of HLA-DR and CD11b
relation of CD36 and CD14
expression of CD13 and CD33;
expression of CD56
Progenitor B cells
enumeration as fraction of total CD34+
VU University Medical Center
Amsterdam The Netherlands
Analysis of a minimized multiparameter panel
% CD34+ myeloblasts
% CD34+ B cells of
total CD34+
CD45 expression of
Ly vs. Myeloblasts
SSC ratio of CD10neutrophils vs. Ly
Ogata et al, Blood, 2006; Ogata/Della Porta, Haematologica, 2009
Della Porta, et al., 2011 (to be submitted); Japan/Italy/Netherlands/Sweden/USA/UK
VU University Medical Center
Amsterdam The Netherlands
Analysis of myelomonopoiesis
April 1, 2010
severe
mild
normal
n=45
n=22
n=58
n=24
n=45
n=34
n=31
n=37
Wells et al., Blood, 2003; Van de Loosdrecht and Westers et al., Blood, 2008
Alhan et al.; unpublished data set june 2010
Chu et al., personal communication ASH2010; Kern et al., Cancer 2010
VU University Medical Center
Amsterdam The Netherlands
FCSS is related to transfusion-dependency and
progression to AML
low/int-I risk MDS
myeloblasts either:
CD5 (o),
CD7 (o) or
CD56 (o) positive
In part published: Loosdrecht AA van de, Westers TM et al., Blood 2008;111
Chu et al., personal communication; ASH#2010
VU University Medical Center
Amsterdam The Netherlands
Clinical impact of flow cytometry in MDS
1. Flow cytometry is recognized in the diagnostic and prognostic
work-up of patients with cytopenia
(ELN-2009; ELN-2011)
2. An ELN consensus guideline for flow cytometry in MDS is now
available
(ELN, Haematologica, 2009; updated 2011; to be submitted)
3. Rationale for clinical use of flow cytometry is defined (ELN-2011:
to be submitted)
VU University Medical Center
Amsterdam The Netherlands
Perspectives: 2011
• Submission 1q/2011: 3 ELN consensus documents
• 4th international ELN meeting in Pavia Italy: co-chair
Matteo dellaPorta (October 2011)
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 diagnostics
 FCSS-revised
 erythroid-lineage
 incorporate software tools for analysis/statistics
 clinical reporting
VU University Medical Center
Amsterdam The Netherlands