Serum CD44 levels predict survival in patients with low-risk myelodysplastic syndromes  J. Loeffler-Ragg, U. Germing, W.R. Sperr, H. Herrmann, H. Zwierzina, P. Valent, H. Ulmer, R. Stauder  Critical Reviews in Oncology / Hematology  Volume 78, Issue 2, Pages 150-161 (May 2011) DOI: 10.1016/j.critrevonc.2010.05.008 Copyright © 2010 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Expression of CD44 on bone marrow leukocytes in a patient with MDS (RAEB-II). Bone marrow leukocytes were examined by multicolor flow cytometry using antibodies against CD34, CD38, and CD45. CD44 was found to be expressed on all CD45+ leukocyte subsets (left upper image), including monocytes (right upper image) as well as immature CD34+/CD38− stem cells (left lower image) and more mature CD34+/CD38+ progenitor cells (right lower image), without major differences in staining intensities. Open histograms (grey line) represent the isotype control. Critical Reviews in Oncology / Hematology 2011 78, 150-161DOI: (10.1016/j.critrevonc.2010.05.008) Copyright © 2010 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Differential expression of serum soluble CD44s (solCD44s) in MDS patients as compared to controls. MDS patients are categorized according to the FAB classification (A) and the WHO classification (B). Boxplots represent 25th and 75th percentiles as box. The bar indicates the median value. Median levels of controls and patients were compared using the Mann–Whitney U-test, *=p<0.05, **=p<0.001. Patient group “nfc” includes RA and RARS patients according to FAB classification that are “not further classifiable” according to WHO criteria. For display purposes two outliers, patients with ID1 (7961ng/ml; (A) and (B)) and ID3 (2773ng/ml; (A)), are not illustrated. Critical Reviews in Oncology / Hematology 2011 78, 150-161DOI: (10.1016/j.critrevonc.2010.05.008) Copyright © 2010 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 Correlation between solCD44s and age: r=0.161, p=0.05 (A); months of survival: r=−0.361, p=0.001 (B); leukocyte counts: r=0.296, p=0.001 (C); monocyte counts: r=0.449, p<0.001 (D). For display purposes two outliers, patient with ID3 (2773ng/ml; (A)) and patient with ID1 (7961ng/ml; (A–D)), are not illustrated. Critical Reviews in Oncology / Hematology 2011 78, 150-161DOI: (10.1016/j.critrevonc.2010.05.008) Copyright © 2010 Elsevier Ireland Ltd Terms and Conditions

Fig. 4 Levels of solCD44s are significantly correlated with overall survival in MDS patients classified according to the FAB* (A) or the WHO* classification (B). The cut-off level (688.5ng/ml) was calculated on the basis of the median of healthy controls plus two standard deviations. Moreover, the correlation between AML-free survival and solCD44s levels was calculated within the WHO-classified MDS patient cohort including CMML (C). For this analysis sAML was defined by bone marrow blasts>19% and the time of the dysplastic phase or observation period was integrated. *Patients with sAML, stem cell transplantation or incomplete observation period were excluded in calculations shown in (A) and (B). In addition, (B) does not include CMML patients. Critical Reviews in Oncology / Hematology 2011 78, 150-161DOI: (10.1016/j.critrevonc.2010.05.008) Copyright © 2010 Elsevier Ireland Ltd Terms and Conditions

Fig. 5 Survival in MDS patients based on the International Prognostic Scoring System (IPSS) (A). The correlation between solCD44s and overall survival is shown in the various IPSS risk subgroups in detail: IPSS low-risk (B), IPSS Int-1 (C), IPSS Int-2 (D) and IPSS high-risk (E). *Patients with sAML, stem cell transplantation or incomplete observation period were excluded. Critical Reviews in Oncology / Hematology 2011 78, 150-161DOI: (10.1016/j.critrevonc.2010.05.008) Copyright © 2010 Elsevier Ireland Ltd Terms and Conditions