VU University Medical Center Amsterdam The Netherlands 3 rd International ELN Workshop:...

VU University Medical CenterAmsterdam The Netherlands

3rd International ELN Workshop:Standardization of Flow Cytometry in MDS

London, Greenwich 4-6 November 2010

a.a. van de loosdrecht, chairr. ireland, co-chair

update ELNet wp8; ASH Orlando, December 4, 2010and ELNet WP conference, Mannheim, February 1, 2011


• 12 countries; 28 participants; in- and outside Europe

• 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

3rd International ELN Workshop:Standardization of Flow Cytometry in MDS

London, Greenwich 4-6 November 2010a.a. van de loosdrecht, chair

r. ireland, co-chair


• define minimal flow cytometric criteria to analyse MDS ELN 2011 [2 consensus documents: rationale/technical]

• Define: role of FCM:– Diagnosis– Prognosis

– predictive value for R/ [emerging new drugs]

– disease monitoring

Goal:


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


Overview of some recommended flow cytometric analyses of subsets in the BM by the ELNet; London 2010

BONE MARROW SUB SET RECOMMENDED ANALYSIS

Erythroid compartment

% of nucleated erythroid cells

Relation CD71 and CD235a

Expression of CD71

Immature myeloid and monocytic progenitors

% of cells

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+


Analysis of a minimized multiparameter panel

% CD34+ myeloblasts

% CD34+ B cells of total CD34+

CD45 expression ofLy vs. Myeloblasts

SSC ratio of CD10-

neutrophils 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


n=45 n=22 n=58 n=24 n=45 n=34 n=31 n=37

Analysis of myelomonopoiesis

Wells et al., Blood, 2003; Van de Loosdrecht and Westers et al., Blood, 2008Alhan et al.; unpublished data set june 2010

Chu et al., personal communication ASH2010; Kern et al., Cancer 2010

severe

mild

normal

April 1, 2010


In part published: Loosdrecht AA van de, Westers TM et al., Blood 2008;111

Chu et al., personal communication; ASH#2010

myeloblasts either:

CD5 (o),CD7 (o) orCD56 (o) positive

FCSS is related to transfusion-dependency and progression to AML

low/int-I risk 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)

Clinical impact of flow cytometry in MDS


Perspectives: 2011• Submission 1q/2011: 3 ELN consensus documents

• 4th international ELN meeting in Pavia Italy: co-chair Matteo dellaPorta (October 2011)

diagnostics FCSS-revised erythroid-lineage incorporate software tools for analysis/statistics clinical reporting

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