Leukemia (1).ppt and bio marker and cbc

leukemia

1. Week 10: Leukemia
AML
ALL
FAB classification
WHO classification
Cytochemistries
MPO
SBB
PAS
ORO
CD markers
Flowcytometry
CML
CLL
Karyotype
Ph chromosome
LAP

2. Signs and Symptoms of AML
Insidious nonspecific onset
Pallor due to anemia
Febrile due to ineffective WBC
Petechiae due to thrombocytopenia
Mucus membrane and gum bleed in
M4 and M5
Bone pain

3. Typical Labs
of AML
Leukocytosis
Blastemia
Leukemic hiatus
Auer rods in M2, M3,
M4
Thrombocytopenia
Anemia
>20% blasts in BM

4. Other Findings
CD 13 and CD 33 in flowcytometry
Cytochemistries
Myeloperoxidase
Sudan black B
Choloroacetate esterase (specific)
Nonspecific esterase

5. FAB (1976) Classification
M0 --Undifferentiated AML
M1 --AML without maturation
M2 --AML with maturation
M3 --Acute Promyelocytic Leukemia
M4 --Acute Meylomonocytic Leukemia
M5 --Acute Monocytic Leukemia
M6 --Erythroleukemia (DiGuglielmo’s)
M7 --Megakaryoblastic Leukemia

7. M1 and M2

10. Myeloperoxidase
(MPO)
p-Phenylene diamine + Catecol + H
2O
2
MPO
> Brown black deposits

11. M3
M5
M4

12. Chloracetate (Specific) Esterase
Myeloid Cell Line
Naphthol-ASD-chloracetate
CAE
> Free naphthol compounds
+ Stable diazonium salt (eg, Fast Corinth)
> Red deposit

13. Non-Specific Esterase
Monocytic Line
Naphthyl acetate
ANAE
> Free naphthyl compounds
+Stable diazonium salt (eg, Fast blue RR)
> Brown deposits

14. Double Esterase in M4
NSE with Fl inhibition
Histiocyte

15. FAB vs WHO Classifications of
Hematologic Neoplasm
FAB criteria
Morphology
Cytochemistry
WHO criteria
Morphology
Immunophenotyping
Genetic features
Karyotyping
Molecular testing
Clinical features

16. WHO Classification of AML
AML with recurrent cytogenic
translocations
AML with multi-lineage dysplasia
AML and myelodysplasia, therapy related
AML, not otherwise categorized

17. AML with Recurrent Cytogenetic
Translocations (WHO 1995)
t(8;21) --some maturation of neutrophilic line;
rare in older patients; AML1/ETO fusion protein;
>90% FAB M2
t(15;17) --APL (granular and microgranular
variants); retinoic acid receptor (RAR) leukemias;
middle aged adults; DIC
inv(16) or t(16;16) --monocytic and granulocytic;
abnormal eosinophilic component
11q23 --monocytic; children; most common is
t(9;11)

19. FAB Classification of ALL
L1: Small homogeneous blasts; mostly in
children
L2: Large heterogeneous blasts; mostly in
adults
L3: “Burkitt” large basophilic B-cell blasts
with vacuoles

20. L3L2

21. Periodic Acid Schiff
Periodic acid + Glycogen
oxidation
> Aldehyde + Schiff reagent
(para-rosaniline, Na metabisulfite)
> Red deposit

22. ALL Cytochemistries
Oil Red O: stains L3 vacuoles
Terminal deoxynucleotidyl transferase
(Tdt): DNA polymerase in early
lymphoblasts
Cell surface markers(CD’s)
Cytoplasmic and surface immunoglobulins:
B-cell line
T-cell receptor (TCR)

23. WHO Classification of
Lymphoproliferative Syndromes
Precursor B Lymphoblastic Leukemia/Lymphoma
(ALL/LBL) --ALL in children (80-85% of
childhood ALL); LBL in young adults and rare;
FAB L1 or L2 blast morphology
Precursor T ALL/LBL --15% of childhood ALL
and 25% of adult ALL
Burkitt Leukemia/Lymphoma (FAB L3)

24. •Antigens•B-Lineage •T-Lineage
•HLA Dr
•Tdt
•CD34
•+
•+
•+
•0 to +
•+
•0
•CD19
•C 22
•CD10
•CD20
•Cyt-
•SIg
•+
•cALL and older
•cALL and older
•Pre-B and older
•Pre-B and older
•B-ALL
•Pre-T
•Pre-T
•Pre-T
•0
•0
•0
•CD7
•CD3
•CD5
•CD2
•CD1
•0
•0
•0
•0
•0
•+
•+
•+
•T-ALL
•T-ALL

25. Indicators Favorable Poor
WBC < 50,000/L 50,000/L
Age 1 -10 < 1 or 10
Gender Female Male
Blast B-cell T-cell and mixed
Karyotype Hyperploidy
Trisomy 4, 10, 17
t(12;21) (TEL/AML1)
Hypoploidy
Trisomy 5
t(1;19 (E2A/PBX1)
Mixed lineage leukemia
T(9;22) (Ph)
BM blast count
during induction
Mkd reduction at day 7Mild reduction at day 7
Prognosis

26. FAB (1982) Classification of
Myeloproliferative Disease (MPD)
Chronic Myelocytic Leukemia (CML)
Polycythemia Vera (PV)
Essential Thrombocythemia (ET)
Agnogenic Myeloid Metaplasia with or without
Myelofibrosis (AMM)
Benign Leukemoid Reaction

27. WHO Classification of MPS
(1997)
CML becomes
CML, Ph + t(9;22) BCR/ABL
Chronic Neutrophilic Leukemia (CNL)
Chronic Eosinophilic Leukemia and Hyper-
eosinophilic Syndrome (CEL/HES)
PV remains PV
ET remains ET
AMM becomes
Chronic Idiopathic Myelofibrosis

28. Myelofibrosis

29. Myelofibrosis

31. Chronic Lymphocytic Leukemia
Exclusive in elderly
Lyphocytosis unrelated to viral infection
Hyper-mature lymphocytes with highly
condensed nuclei
Smudge cells: preventable with a drop of
bovine albumin

32. CLL
PB and BM

36. WHO Lymphoid Neoplasms
B cell neoplasms
T/NK cell neoplasms
Hodgkin lymphoma (disease)

37. Mature B Cell Neoplasms
B cell CLL/SLL
B prolymphocytic
leukemia
Burkitt’s lymphoma /
leukemia
Splenic marginal zone
B lymphoma
Extranodal marginal B
lymphoma
Hairy cell leukemia
Lymphoplasmocytic
leukemia
Mantle cell lymphoma
Plasma cell myeloma /
plasmacytoma
Follicular lymphoma
Diffuse large B
lymphoma

38. T/NK Cell Neoplasms
T prolymphocytic
leukemia
T granular
lymphocytic leukemia
Aggressive NK cell
leukemia
Adult T lymphoma /
leukemia
Mycosis fungoides
(Sezary syndrome)
Anaplastic large cell
lymphoma
Hepatosplenic T
lymphoma
Peripheral T lymphoma
Immunoblastic T
lymphoma