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Flow Cytometric Studies of MDS:

Detailed flow cytometric study on MDS highlighting abnormal CD marker expression in granulocytes and monocytes, aiding in diagnosis. The study examines the dysmaturation features and atypical cells in bone marrow samples.

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Flow Cytometric Studies of MDS:

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  1. Flow Cytometric Studies of MDS: • Increased CD34+ cells (>3% of nonerythroid cells) in MDS BM Kanter-Lewensohn et al. Eur.J., Haematol.56:124-129, 1996 • Decreased CD71 in erythroblasts • Also in anemia of chronic disease Kuiper-Kramer et al 97:127-131, 1997

  2. MOLECULES ANALYZED IN THIS STUDY CD24 (CD52/HSA) CD15 (x hapten, Lewis x) CD10 (NEP) CD13 (APN) CD35 (CR1) CD203 (97A6) CD99 (E2, MIC2) CD14 (LPS receptor) CD36 (Thrombosp. rec) CD56 (NCAM) CD45R0 Tyr phosphatase CD11a b2 integrin CD11b b2 integrin CD11c b2 integrin CD43 Sialomucin CD33 (Sialoadhesin) CD64 (Fc receptor) CD16 (Fc receptor) CD66b (CEA family) CD66c (CEA family) Del Vecchio, 2006

  3. GRANULOID CELLS IN NORMAL BONE MARROW the concave pattern CD13 CD11b the right angle CD16 CD16 the reverse right angle the mature population CD10 CD11b CD64 CD15

  4. GRANULOID CELLS IN MDS BONE MARROW the convex pattern CD13 CD11b the disrupted right angle CD16 CD16 the missing population CD10 CD11b the disrupted reverse right angle CD64 CD15

  5. 0 1 2 3 4 10 10 10 10 10 CD11a FITC -> 0 1 2 3 4 10 10 10 10 10 CD11a FITC -> PMN MATURATION PATHWAY CD11a/CD11b/CD45/CD14 last step increase

  6. 0 1 2 3 4 0 10 1 10 2 10 3 10 4 10 10 10 10 10 10 CD43 FITC -> CD43 FITC -> PMN MATURATION PATHWAY CD43/CD11b/CD45/CD14 slow decrease

  7. PMN MATURATION PATHWAY CD99/CD11b/CD45/CD14 0 1 2 3 4 10 0 10 1 10 2 10 3 10 4 10 10 10 10 10 CD99 FITC -> CD99 FITC -> two phases

  8. PMN MATURATION PATHWAY CD66b/CD66c/CD45/CD14 0 1 2 3 4 10 10 10 10 10 0 1 2 3 4 10 10 10 10 10 CD66b FITC -> CD66b FITC -> the star model

  9. PMN MATURATION PATHWAY CD24/CD11c/CD45/CD14 0 1 2 3 4 10 10 10 10 10 0 1 2 3 4 10 10 10 10 10 CD24 FITC -> CD24 FITC -> the anvil model

  10. 0 1 2 3 4 10 10 10 10 10 0 1 2 3 4 10 10 10 10 10 CD35 FITC -> CD35 FITC -> PMN MATURATION PATHWAY CD35/CD11b/CD45/CD14 last step increase

  11. CD14+ monocytes lymphocytes CD14- monoblasts CD11b+ granulocytes CD11b-immature myeloid cells 0 256 512 768 1024 SSC-H -> Monocytes CD36/CD11b/CD45/CD14

  12. grans 0 1 2 3 4 10 10 10 10 10 0 1 2 3 4 10 10 10 10 10 CD14 APC -> CD14 APC -> CYTOMETRIC EVIDENCE OF IMMATURE (CD14-) and ATYPICAL (CD56+) MONOCYTES

  13. ERYTHROID DEVELOPMENT CD71 - CD105 - LDS751 - CD45 1 1 2 2 3 3 1 2 3 proerythro Intermediate mature erythroblasts FWSC CD105

  14. 117 CASES INCLUDED IN THE STUDY Hemolytic Iron deficiency Hypoplastic

  15. Analytical tools to diagnose MDS by FCM

  16. MULTIMARKER SET ANALYSIS CD16-CD11b abn or CD16-CD13 abn or CD64-CD11b abn or >% of My1 gate >% of area2 or >% of CD14+CD56+ or >CD105/FSC abn or >% of CD34+ AND

  17. 100 A RA RCMD RAEB CMML MULTIMARKER SET ANALYSIS OF MDS P<0.001

  18. A APL AML ALL MF PV ET MULTIMARKER SET ANALYSIS OF OTHER CLONAL HEMATOLOGICAL DISEASES P<0.001

  19. Della Porta 2006

  20. Ogata, Blood 2006

  21. Denise Wells, 2003

  22. Flow vs No-Flow ST 30’ 3-3

  23. CD105 PE -> CD19 -> SSC-H -> XY HOSPITAL Immunology Unit Cytometric typing of MDS First name:Xxxxxxxxx Last Name:Yyyyyyyyy Date of birth: dd.mm.yyyy Case number: 2412 Division: Hematology Specimen: Bone Marrow First obs:_X_Follow up:___ Diagnostic suspect: MDS. Antigen studied: CD45, CD16, CD11b, CD13, CD64, CD56, CD19, CD71, CD105,CD34, CD33, CD14. Technique: multicolor immunofluorescence Bone marrow aspirate characterized by abundant cellularity (50000 cells/ml). Granulocytopoiesis shows dysmaturation features, with marked alteration of CD16/CDC11b, CD16/CD13 and CD64/CD11b patterns. Presence of atypical CD56+ monocytes. Normal expression of CD71 on erythroblasts. Normal erythroid maturation pattern assessed by FWSC-CD105. Takentogether, the BM characterization suggests the condition of dysgranulo-dysmonopoiesis. Date of typing Signature

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