CLINICAL PATHOLOGY UPDATE on SURAMADE ISurabaya, 15th Juliy 2011                            YETTI HERNANINGSIH, dr., SpPK ...
(Gorczyca W, Emmons FM, 2008)                                2
Flow cytometric immunophenotyping is          important for the          Distinction between ALL and AML,        identific...
SAMPLE STAINING         1.SURFACE           ANTIGEN         2.INTRACELLULAR           ANTIGEN (TdT,           cCD3, cCD22,...
5
6(Gorczyca W, Emmons FM, 2008)
Immunophenotyping of AML(Campana & Behm,2000)                           7
(Courtesy Sayed DM)                  8
9
Figure 6.22 Schematic illustration of antigen expression during normal T-cell and NK-cellmaturation. Note that, although t...
(Concise Manual of Hematology and Oncology, 2008)                                                    11
(Concise Manual of Hematology and Oncology, 2008)                                                    12
Biphenotypic Acute Leukemia If > 2 points is scored for both myeloid and one of the  lymphoid lineages the case is classi...
Undifferentiated Acute Leukemias Rare subgroup of acute leukemias which cannot be  further classified using the above men...
ABERRANTImmature B cell   Abberant : co express an antigen of                  different lineage e.g. CD 65 (myeloid      ...
How to store and send sample for         Immunophenotyping              • Bone marrow (BM) aspirate orSpecimen        Whol...
How to store and send sample for       ImmunophenotypingHemogram   • Should be accompanied             (if present)Schedul...
Panel of antibodies recommended by European Group for theImmunoloogical Characterization of aleukemia (EGIL) fordiagnosis ...
Application of Immunophenotyping      at Dr. Soetomo Hospital CD7 FITC/CD33 PE/CD45 PerCPHLA-DR FITC/CD13 PE/CD45 PerCP CD...
20
21
Positive expression :  membran > 20%  cytoplasmic >10%                        22
Dr. SOETOMO HOSPITAL      EXPERIENCES                       23
Mrs.Sh, 44 y.o.                  24
CD13, CD33, Anti-HLA-DR, CD34  POSITIVE               Conclusion:             Myeloid Lineage                            ...
Mrs.Sm, 30 y.o.                  26
CD13, Anti-HLA-DR, CD33, CD10  POSITIVE           CML Conclusion: Myeloid Lineage Aberrant Expression CD10               ...
Mrs.Sl, 27 y.o.                  28
CD33, CD13, Anti-HLA-DR, CD34, CD3, CD10, CD19                      POSITIVE                    Conclusion:Suspected Biph...
Mrs.M, 60 y.o.                 30
AML-M5bCD33, CD13, Anti-HLA-DR  POSITIVE            Conclusion:         Myeloid Lineage                                  ...
Mr.Ms, 24 y.o.                 32
Anti-HLA-DR, CD10, CD19, CD34, CD3, CD20                  POSITIVE                  Conclusion:B-Lymphoid Lineage with Ab...
Mr.My, 51 y.o.                 34
CD33, CD7, CD13, Anti-HLA-DR, CD34  POSITIVE                  Conclusion: Myeloid Lineage with Aberrant Expression CD7   ...
WBC : 10.00 (90.8% Lymphocyte)HGB : 7.2 23 y.o.   Mr.Y,PLT : 7                                 36
Anti-HLA-DR, CD10, CD19, CD34, CD20             POSITIVE            Conclusion:        B-Lymphoid Lineage                ...
Mr.I, 24 y.o.                38
CD33, CD13, Anti-HLA-DR, CD10, CD19, CD34                  POSITIVE                 Conclusion:Suspected Biphenotypic (My...
References Nguyen D, Diamond L.W, Braylan R.C. 2007. Flow  Cytometry in Hematopathology. 2nd ed. Humara Press.  New Jerse...
THANK YOU
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  1. 1. CLINICAL PATHOLOGY UPDATE on SURAMADE ISurabaya, 15th Juliy 2011 YETTI HERNANINGSIH, dr., SpPK Clinical Pathology Department Dr. Soetomo Hospital Surabaya
  2. 2. (Gorczyca W, Emmons FM, 2008) 2
  3. 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. 4. SAMPLE STAINING 1.SURFACE ANTIGEN 2.INTRACELLULAR ANTIGEN (TdT, cCD3, cCD22, MPO) 3.DNA CONTENT 4
  5. 5. 5
  6. 6. 6(Gorczyca W, Emmons FM, 2008)
  7. 7. Immunophenotyping of AML(Campana & Behm,2000) 7
  8. 8. (Courtesy Sayed DM) 8
  9. 9. 9
  10. 10. Figure 6.22 Schematic illustration of antigen expression during normal T-cell and NK-cellmaturation. 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. 11. (Concise Manual of Hematology and Oncology, 2008) 11
  12. 12. (Concise Manual of Hematology and Oncology, 2008) 12
  13. 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. 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. 15. ABERRANTImmature 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. 16. How to store and send sample for Immunophenotyping • Bone marrow (BM) aspirate orSpecimen Whole blood (WB) with EDTA • No blood clot or hemolysisStorage and • At room temperature (never be Transport transported on dry ice) • 24 hours, send to lab as soon as Stability possible 16
  17. 17. How to store and send sample for ImmunophenotypingHemogram • Should be accompanied (if present)Schedule • Every working day Report • 1 working days issues 17
  18. 18. Panel of antibodies recommended by European Group for theImmunoloogical Characterization of aleukemia (EGIL) fordiagnosis and classificatin of acute leukemia (1995) (Courtesy Sayed DM) 18
  19. 19. Application of Immunophenotyping at Dr. Soetomo Hospital CD7 FITC/CD33 PE/CD45 PerCPHLA-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. 20
  21. 21. 21
  22. 22. Positive expression : membran > 20% cytoplasmic >10% 22
  23. 23. Dr. SOETOMO HOSPITAL EXPERIENCES 23
  24. 24. Mrs.Sh, 44 y.o. 24
  25. 25. CD13, CD33, Anti-HLA-DR, CD34  POSITIVE Conclusion: Myeloid Lineage 25
  26. 26. Mrs.Sm, 30 y.o. 26
  27. 27. CD13, Anti-HLA-DR, CD33, CD10  POSITIVE CML Conclusion: Myeloid Lineage Aberrant Expression CD10 27
  28. 28. Mrs.Sl, 27 y.o. 28
  29. 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. 30. Mrs.M, 60 y.o. 30
  31. 31. AML-M5bCD33, CD13, Anti-HLA-DR  POSITIVE Conclusion: Myeloid Lineage 31
  32. 32. Mr.Ms, 24 y.o. 32
  33. 33. Anti-HLA-DR, CD10, CD19, CD34, CD3, CD20  POSITIVE Conclusion:B-Lymphoid Lineage with Aberrant Expression CD3 CD13: 13.9% 33
  34. 34. Mr.My, 51 y.o. 34
  35. 35. CD33, CD7, CD13, Anti-HLA-DR, CD34  POSITIVE Conclusion: Myeloid Lineage with Aberrant Expression CD7 35
  36. 36. WBC : 10.00 (90.8% Lymphocyte)HGB : 7.2 23 y.o. Mr.Y,PLT : 7 36
  37. 37. Anti-HLA-DR, CD10, CD19, CD34, CD20  POSITIVE Conclusion: B-Lymphoid Lineage 37
  38. 38. Mr.I, 24 y.o. 38
  39. 39. CD33, CD13, Anti-HLA-DR, CD10, CD19, CD34  POSITIVE Conclusion:Suspected Biphenotypic (Myeloid + B-Lymphoid) Acute Leukemia 39
  40. 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. 40
  41. 41. THANK YOU

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