mixed- LAB SKILLS- YR 3- LEUKEMIA CLASSIFICATION- Optimized- old
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Transcript of mixed- LAB SKILLS- YR 3- LEUKEMIA CLASSIFICATION- Optimized- old
CLASSIFICATION OF THE LEUKEMIASCLASSIFICATION OF THE LEUKEMIAS
ACUTE• ACUTE MYELOID
LEUKEMIA
• ACUTE LYMPHOBLASTIC LEUKEMIA
CHRONIC• CHRONIC MYELOCYTIC
LEUKEMIA
• CHRONIC LYMPHOCYTIC LEUKEMIA
ACUTE LEUKEMIA CLASSIFICATIONACUTE LEUKEMIA CLASSIFICATION
• Morphology Morphology • CytochemistryCytochemistry• Immunophenotyping: Immunophenotyping: identifying antigens identifying antigens associated with cells (cell markersassociated with cells (cell markers))• Cytogenetics and molecular markersCytogenetics and molecular markers
ACUTE LEUKEMIA ACUTE LEUKEMIA CLASSIFICATION SYSTEMS CLASSIFICATION SYSTEMS
• FAB (French-American-British)
• MIC (Morphologic, Immunophenotypic and Cytogenetic)
• WHO
THE FRENCH-AMERICAN-BRITISH (FAB) THE FRENCH-AMERICAN-BRITISH (FAB) CLASSIFICATION OF THE ACUTE LEUKEMIASCLASSIFICATION OF THE ACUTE LEUKEMIAS
• Depends on a combination of MORPHOLOGIC, CYTOCHEMICAL
• for some categories, IMMUNOPHENOTYPIC characteristics
A Flow CytometerA Flow Cytometer
FAB CLASSIFICATIONFAB CLASSIFICATION
ACUTE MYELOID LEUKEMIAS
• M0: undifferentiated• M1: without maturation• M2: with granulocytic maturation• M3: promyelocytic• M4: myelomonocytic• M5: monocytic• M6: erythroleukemia• M7: megakaryoblastic
AML- FAB classificationAML- FAB classificationMYELOID /
GRANULOCYTIC LINEAGE• M0• M1• M2• M3
• MONOCYTIC +/- GRANULOCYTIC
• M4• M5
ERYTHROID• M6
MEGAKARYOCYTIC• M7
• CHRONIC MYELOID LEUKEMIA
CML: BLOOD SMEARCML: BLOOD SMEAR
CML-BMCML-BM
CML: BLOOD SMEARCML: BLOOD SMEAR
CML CML (courtesy www)(courtesy www)
CML BASOPHILSCML BASOPHILS
CML ; MARROWCML ; MARROW
ACUTE LEUKEMIAACUTE LEUKEMIA
FAB CLASSIFICATIONFAB CLASSIFICATION
ACUTE LYMPHOBLASTIC LEUKEMIAACUTE LYMPHOBLASTIC LEUKEMIA• L1L1: Uniform small size, high N/C ratio, regular nuclear
margin, inconspicuous nucleoli
• L2L2: Heterogeneous large and smaller cells, variable cytoplasm and nuclear margin, prominent nucleoli
• L3L3: Large cells, deeply basophilic and vacuolated cytoplasm, prominent nucleoli
ACUTE LEUKEMIA- BLASTSACUTE LEUKEMIA- BLASTS
Acute leukemia (courtesy www)Acute leukemia (courtesy www)
AML- BLASTS WITH AUER RODSAML- BLASTS WITH AUER RODS
Myeloperoxidase reaction in blastsMyeloperoxidase reaction in blasts
AML: CAEAML: CAE
ANAE [AML-M7]ANAE [AML-M7]
AML: AUER RODSAML: AUER RODS
PAS [ALL]PAS [ALL]
AML-M1AML-M1 (courtesy www) (courtesy www)
CYTOCHEMISTRY: REACTIONS IN BLASTSCYTOCHEMISTRY: REACTIONS IN BLASTS
AML• Myeloperoxidase +• Sudan Black +• Chloroactate esterase +• Nonspecific esterase + (M4, M5)• Periodic acid-Schiff - (+ in M6)• Acid phosphatase + (M4,M5,M6)
ALL - - - - + (40-60%cases) + (T-ALL)
IDENTIFYING BLASTS: OTHER CHARACTERISTICSIDENTIFYING BLASTS: OTHER CHARACTERISTICS
• IMMUNOLOGICAL: flow cytometry for ANTIGENS immunohistochemistry
• MOLECULAR: PCR, southern blotting for Ig AND TCR genes
• CYTOGENETIC for chromosomal abnormalities
• ELECTRONMICROSCOPIC
• IMMUNOLOGICAL (ANTIGENIC) MARKERS:
• IMMUNOPHENOTYPE determined by IMMUNOFLOWCYTOMETRY and/or IMMUNOHISTOCHEMISTRY
MYELOID MARKERSMYELOID MARKERS
• Myeloperoxidase• CD13, CD33• CD 11 b/c, CD14, CD15
IMMUNOPHENOTYPE: IMMUNOPHENOTYPE: CELL MARKERS (ANTIGENS)CELL MARKERS (ANTIGENS)
LYMPHOID
• B-LINEAGE- CD79a, CD22, µ chain, CD19, CD10 TdT
• T-LINEAGE- CD3 CD2, CD5, TdT, CD7
CLLCLL• Usually, lymphocytes are > 10 x 109/l with
involvement of bone marrow
• A diagnosis of CLL is possible with lower counts if morphology and immunophenotype are consistent with CLL
• A diagnosis of SLL but not CLL can be made histologically in the absence of BM or blood involvement
• The pattern of marrow infiltration is a guide to prognosis
CLL CLL
IMMUNOPHENOTYPE
• Weak/dim IgM or IgM and IgD• CD5, CD19• CD10-, cyclin D1-
CLL- LYMPHOCYTESCLL- LYMPHOCYTES
CLL- (courtesy www)CLL- (courtesy www)
PRACTICAL USES OF CLASSIFYINGPRACTICAL USES OF CLASSIFYING
• APPROPRIATE THERAPY
• PROGNOSTICATION
• IDENTIFICATION OF MINIMAL RESIDUAL DISEASE