1. VU University Medical Center Amsterdam The Netherlands 3 rd 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

2. VU University Medical Center Amsterdam The Netherlands 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 3 rd 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

3. VU University Medical Center Amsterdam The Netherlands 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:

4. 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

5. VU University Medical Center Amsterdam The Netherlands Overview of some recommended flow cytometric analyses of subsets in the BM by the ELNet; London 2010

6. 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 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

7. VU University Medical Center Amsterdam The Netherlands 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Analysis of myelomonopoiesis Wells et al., Blood, 2003; Van de Loosdrecht and Westers et al., Blood, 2008 Alhan et al.; unpublished data set june 2010 Chu et al., personal communication ASH2010; Kern et al., Cancer 2010 severe mild normal April 1, 2010

8. VU University Medical Center Amsterdam The Netherlands 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) or CD56 (o) positive FCSS is related to transfusion-dependency and progression to AML low/int-I risk MDS

9. VU University Medical Center Amsterdam The Netherlands 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

10. VU University Medical Center Amsterdam The Netherlands Perspectives: 2011  Submission 1q/2011: 3 ELN consensus documents 4 th 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