L38 and L16 Approach to hematological malignancies and investigations Flashcards
Hematological overview:
Acute + Lymphoid malignancy?
Acute Lymphoblastic leukemia (ALL)
Hematological overview:
Chronic + Lymphoid malignancy?
Chronic Lymphoid leukemia (CLL)
(Lymphoproliferative neoplasm)
- Plasma cell neoplasm
Hematological overview:
Acute + Myeloid malignancy?
Acute myeloid leukemia
Hematological overview:
Chronic + Myeloid malignancy?
Chronic myeloid leukemia
Myeloproliferative neoplasm
In AML, there are excessive _________ in the bone marrow, pancytopenia in PB and also increase in WBC.
blast cells
In Myelodysplasitc syndrome, proliferation of cells are normal or excess but _____________.
Blast cells in bone marrow is <20% but hypercellular.
Pancytopenia in PB.
ineffective
differentiation of cells impaired
When is a bone marrow study required? What possibilities are evaulated?
- unexplained pancytopenia (due to BM failure)
1. Acute leukemia
2. Aplastic anemia
3. Infiltration: TB, carcinoma
What is the main difference between BM aspiration and biopsy?
Aspiration:
- cytological abnormalities
Biopsy:
- histological abnormalities: architecture, infiltration, cellularity
Immunophenotyping: Flow cytometry is used to? (2)
- Determine lineage
2. Detect minimal residual diseases (MRD)
What is Forward scatter and Side scatter detecting?
Forward scattering: Cell size
Side scattering: Granularity
Granulocytes / Neutrophils FS and SS?
Large and more granular, therefore with high SS and FS
Monocytes FS and SS?
Large cells but not so granular, therefore high FS but low SS.
Lymphocytes FS and SS?
SMall cells, not granular, therefore low FS and low SS.
Gating means to isolate a single population of interest from a heterogeneous sample by its specific ____________ and SS/FS properties.
specific antigen
e.g. CD45
Which types of cells have bright and dim CD45 antigen expression respectively?
Bright: Mature leukocytes
Dim: Blast
Negative: erythroid and non-hematological cells
What is CD in CD45.
Cluster differentiation, antigens found on leukocytes
Which of the following is not a B-lymphoid marker? A. CD19 B. CD20 C. CD22 D. CD33 E. CD79a F. CD10
D
Myeloid marker!
Which of the following is not a T-lymphoid marker? A. CD2 B. CD3 C. CD5 D. CD7 E. CD13
E
Myeloid marker!
Which of the following is not a myeloid marker? A. CD33 B. CD13 C. CD117 D. CD34 E. Myeloperoxidase (MPO)
D
in hematopoietic progenitor cells
Which of the following are hematopoietic progenitor markers?
A. CD34
B. TdT
C. HLA-DR
All of the above
In patient with paroxysmal nocturnal hemoglobinuria (PNH), what markers are used to monitor the RBCs and neutrophil levels respectively? Levels of them increases/decreases.
RBC: CD59
Neutrophils: FLAER;
decreases
In HS (hereditary spherocytosis), marker ____________is reduced.
EMA
= eosin-5′-maleimide
To assess cytogenetics, what techniques can be used? (2) Briefly describe each of them.
- G-banding: Identify extra or missing chromosome and structural aberrations in KARYOTYPE, e.g. PML-RARA in APL
- FISH: Target specific aberrant DNA sequence, e.g. PML-RARA
To do molecular study, PCR, RT-PCR, RQ-PCR, NGS etc. can be done to detect?
Point mutations, insertions or deletions that are not detectable in cytogenetics
e.g. FLT3-ITD in AML
+
MRD
What is MRD (minimal residual disease)?
Small number of __________ that remain in the patient during/after treatment
leukemic cells
In traditional assessment of treatment response monitoring by morphology, e.g. PB count, BM cellularity, BM blast count for remission…, what are the 2 main issues?
- Not sensitive: patients in morphological complete remission may still have a large number of residual leukemia cells
- Haematogones: normal B cell progenitors highly resemble residual leukemic cells; higher haematogones in BM after temporary stop in chemotherapy, end of treatment/ HSCT transplantation
RQ-PCR is one of the ways of MRD detection. In patients with B-ALL and T-ALL, what exactly is measured?
B-ALL: Clonal rearrangement of Ig gene
T-ALL: TCR
RQ-PCR can also measure expression of gene fusion and leukemia specific markers.
What is detected in the following disease?
1. CML
2. APL
3. AML
- CML: BCR-ABL
- APL: PML-RARA
- AML: NPM1 mutation
Compared to flow cytometry, RQ-PCR is more/less sensitive and more/less time-consuming.
More sensitive;
More time consuming
Flow cytometry is used to distinguish ___________ from ___________ in leukemia-associated immunophenotype.
Leukemic cells from hematogones (normal prongenitor cells)
Example: Some B lymphoid cells aberrantly expressed CD-5, a __________ marker. This is a feature of _________.
T-linieage marker;
CLL
Example: CLL: CD20+ but CD10-. CD10 is usually +ve in ?
follicular lymphoma (B cell proliferative neoplasm)
PNH cells showed _________ reduction. Lost of it from red cells allows uncontrolled ___________activation and intravascular hemolysis.
CD59;
complement
