1. CLINICAL PATHOLOGY UPDATE on SURAMADE I
Surabaya, 15th Juliy 2011
YETTI HERNANINGSIH, dr., SpPK
Clinical Pathology Department
Dr. Soetomo Hospital Surabaya

2. (Gorczyca W, Emmons FM, 2008)
2

3. Flow cytometric immunophenotyping is
important for the
Distinction between ALL and AML,
identification of B-cell or T-cell lineage
Assessing response to treatment, including
the detection of MRD
Detection aberrant expression
3

4. SAMPLE STAINING
1.SURFACE
ANTIGEN
2.INTRACELLULAR
ANTIGEN (TdT,
cCD3, cCD22,
MPO)
3.DNA CONTENT
4

5. 5

6. 6
(Gorczyca W, Emmons FM, 2008)

7. Immunophenotyping of AML
(Campana & Behm,2000)
7

8. (Courtesy Sayed DM)
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9. 9

10. Figure 6.22 Schematic illustration of antigen expression during normal T-cell and NK-cell
maturation. Note that, although the common lymphoid stem cell presumably arises in the bone
marrow, most T-cell maturation occurs in the thymus. As with the B cells (Fig. 6.12), T-cell
activation occurs after exposure to antigen.(Courtesy Dr. Dan Sabath.)
10
(Tkachuk DC, Hirschmann JV, 2007)

11. (Concise Manual of Hematology and Oncology, 2008)
11

12. (Concise Manual of Hematology and Oncology, 2008)
12

13. Biphenotypic Acute Leukemia
 If > 2 points is scored for both myeloid and one of the
lymphoid lineages the case is classified as biphenotypic 13

14. Undifferentiated Acute Leukemias
 Rare subgroup of acute leukemias which cannot be
further classified using the above mentioned criteria.
 Usually CD34+, HLA-DR+, CD38+, and CD7+.
14

15. ABERRANT
Immature B cell Abberant : co express an antigen of
different lineage e.g. CD 65 (myeloid
antigen) expressed on an immature
B cell (CD19+, CD34+, cyCD22+,
cyCD79a+, TdT+) 15

16. How to store and send sample for
Immunophenotyping
• Bone marrow (BM) aspirate or
Specimen Whole blood (WB) with EDTA
• No blood clot or hemolysis
Storage and • At room temperature (never be
Transport transported on dry ice)
• 24 hours, send to lab as soon as
Stability possible
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17. How to store and send sample for
Immunophenotyping
Hemogram • Should be accompanied
(if present)
Schedule • Every working day
Report
• 1 working days
issues
17

18. Panel of antibodies recommended by European Group for the
Immunoloogical Characterization of aleukemia (EGIL) for
diagnosis and classificatin of acute leukemia (1995)
(Courtesy Sayed DM) 18

19. Application of Immunophenotyping
at Dr. Soetomo Hospital
CD7 FITC/CD33 PE/CD45 PerCP
HLA-DR FITC/CD13 PE/CD45 PerCP
CD19 FITC/CD10 PE/CD45 PerCP
CD3 FITC/CD34 PE/CD45 PerCP
CD5 FITC/CD20 PE/CD45 PerCP
19

20. 20

21. 21

22. Positive expression :
membran > 20%
cytoplasmic >10%
22

23. Dr. SOETOMO HOSPITAL
EXPERIENCES
23

24. Mrs.Sh, 44 y.o.
24

25. CD13, CD33, Anti-HLA-DR, CD34  POSITIVE
Conclusion:
Myeloid Lineage
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26. Mrs.Sm, 30 y.o.
26

27. CD13, Anti-HLA-DR, CD33, CD10  POSITIVE
CML Conclusion:
Myeloid Lineage Aberrant Expression CD10
27

28. Mrs.Sl, 27 y.o.
28

29. CD33, CD13, Anti-HLA-DR, CD34, CD3, CD10, CD19 
POSITIVE
Conclusion:
Suspected Biphenotypic (Myeloid + B-Lymphoid) Acute
Leukemia with Aberrant expression CD3
29

30. Mrs.M, 60 y.o.
30

31. AML-M5b
CD33, CD13, Anti-HLA-DR  POSITIVE
Conclusion:
Myeloid Lineage
31

32. Mr.Ms, 24 y.o.
32

33. Anti-HLA-DR, CD10, CD19, CD34, CD3, CD20
 POSITIVE
Conclusion:
B-Lymphoid Lineage with Aberrant Expression CD3
CD13:
13.9%
33

34. Mr.My, 51 y.o.
34

35. CD33, CD7, CD13, Anti-HLA-DR, CD34  POSITIVE
Conclusion:
Myeloid Lineage with Aberrant Expression CD7
35

36. WBC : 10.00 (90.8% Lymphocyte)
HGB : 7.2 23 y.o.
Mr.Y,
PLT : 7
36

37. Anti-HLA-DR, CD10, CD19, CD34, CD20
 POSITIVE
Conclusion:
B-Lymphoid Lineage
37

38. Mr.I, 24 y.o.
38

39. CD33, CD13, Anti-HLA-DR, CD10, CD19, CD34
 POSITIVE
Conclusion:
Suspected Biphenotypic (Myeloid + B-Lymphoid)
Acute Leukemia
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40. References
 Nguyen D, Diamond L.W, Braylan R.C. 2007. Flow
Cytometry in Hematopathology. 2nd ed. Humara Press.
New Jersey.
 Gorczyca W, Emmons F.N. 2008. Atlas of Differential
Diagnostic in Neoplastic Hematopathology. 2nd ed.
Genzyme Genetics. New York.
 Tkachuk D.C, Hirschmann J.V. 2007. Wintrobe Atlas of
Clinical Hematology. 1st ed. Lippincott Williams & Wilkins.
 Berger DP, Engelhand m, Henβ H, Mertlesmann
(eds.).2008. Concise Manual Hematology and Oncology.
Springer. Heidelberg.
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41. THANK
YOU