Acute Leukaemia

H.J.Ruwindi Lakmina Silva

BSc in Medical Laboratory Sciences (KDU)
Reading for PhD (UOK)
Leukaemia

• It is a hematological malignancy caused due to abnormal proliferation

of white blood cells and their progenitor cells.

• Healthy haemopoietic cells get replaced by abnormal leukemia cells.

• Extramedullary haemopoiesis occurs.

• First originate from bone marrow and then invade lymph nodes,
spleen and liver.
Classification of leukaemia

Leukaemia

Acute leukaemia Chronic leukaemia

Acute Myeloid Chronic Myeloid

Leukaemia Leukaemia
(AML) (CML)

Chronic
Acute Lymphoid
Lymphoid
Leukaemia (ALL)
Leukaemia (CLL)
 Laboratory Diagnosis of Leukemia

1. Full blood count

Hb= Reduced
WBC = Markedly increased/normal/decreased
Platelet = varies : acute= low, chronic = normal or high
2. PBS
RBC = Normocytic & Normochromic
WBC = Marked/Moderate/Mild increased. Abnormal cells are present.
Platelet= Varies (low to high)
3. Bone marrow biopsy
- Abnormal cells
- Cellularity: erythropoiesis/ granulopoiesis/ thrombopoiesis
4.Trephine biopsy
- In order to examine both bone and marrow abnormalities
- Detailed structural abnormalities could not be examined
5. Cytochemical stains
- Differential diagnosisof leukaemia can be done using special cytochemical
stains (Sudan black B, Periodic Acid Shiff, Neutrophils alkaline phosphatase,etc..).
6. Flowcytometric immunophenotyping

- Leukemic cell surface antigens are identified by using monoclonal antibodies.

- Use to confirm the type of leukaemia.

(CD 13, CD 19, CD20,etc..)

7. Genetic analysis

-Cytogenetic and molecular analysis to determine mutations.

Treatment

• Chemotherapy

• Radiotherapy

• Bone marrow transplantation

• Gene therapy
 Acute Leukaemias

• Acute leukaemia is normally defined as the presence of over 20% (> 20%) of

blast cells in the bone marrow.

•Acute leukaemias are aggressive maliganancies.

•Genetic mutations cause malignant transformation in haemopoietic stem

cells.
•This results in:
• Increased rate of cell proliferation
• Impaired cellular differentiation
 Acute Myeloid Leukaemia
• AML is the most common acute leukaemia in adults.

• It forms only a minor fraction (10–15%) of the leukaemias in childhood.

• Myeoloid lineage is affected resulting in accumulation of myeloblasts in the bone marrow

replacing healthy haemopoietic cells.

• The AML genome contains an average of about 10 mutations within protein‐coding

genes.

• Myeloblasts are not differentiated in to granulocytes.

• Myeloblasts will transfered in to the blood and to other organs.

Signs and Symptoms

• Anemia

• Bleeding gums

• Fatigue

• Recurrent infections

• Weight loss

• Fever
Classification of AML
Six main groups:
1. AML with recurrent genetic abnormalities
2. AML with myelodysplasia‐related changes
3. Therapy‐related myeloid neoplasms(t‐AML)
4. AML, not otherwise specified

5. Myeloid sarcoma

6. Myeloid proliferations related to Down’s

syndrome
 Laboratory diagnosis of AML

1. Full blood count

Hb= Reduced
WBC = Markedly increased
Platelets = low
*DC= Increased blasts cells
2. PBS
RBC - Normocytic & normochromic.
WBC - Marked leucocytosis with many myeloblasts and/or abnormal monocytes
Platelets – low
3. Bone marrow

• Hypercellular

• Numerous blasts cells (myeloblasts)

4. Cytochemical staining

• Sudan black B: stain lipid granules in myeloblasts

• Myeloperoxidase

• Double esterase
Sudan black B positive Myeloperoxidase positive Double esterase positive
5. Immunophenotyping

• CD13, CD33, CD117

• CD11C, CD64, CD14 = + Monocytic

6. Cytogenetic and molecular genetic analysis

“Auer rods”
Abnormal promyelocytes Abnormal monocytoid cells

BM: abnormal large monocytoid blast like cells

 Acute lymphoblastic leukaemia (ALL)
•ALL is the most common malignancy in childhood accounting for nearly 1/3 of all

childhood cancers.

•It can also be seen in adults.

•Genetic mutation (chromosomal translocation) affect the B or T cell lymphoid

lineage causing proliferation of lymphoblasts that replace healthy haemopoietic

cells in the bone marrow.

•This leads to anemia, neutropenia and thrombocytopenia.

•Abnormal lymphoblasts are then transferred to blood stream and to other vital
 French-American-British (FAB) classification

• Based on the morphology

1. ALL-L1

2. ALL-L2

3. ALL-L3
ALL-L1 ALL-L2
• Agranular small lymphoblasts • Two types of blasts cells >20%:
>20% Small lymphoblasts

• Aure rods are not frequently Large lymphoblasts

• Aure rods are not frequently found
found

ALL-L3
• Lymphoblasts >30%
• Vacuolated cytoplasm in
lymphoblasts: “Burkitt cells”
ALL-L1 ALL-L2 ALL-L3
 Laboratory diagnosis of ALL
1. Full blood count

Hb= Reduced

WBC = Markedly increased/ normal/ decreased

Platelets = low
• *DC= Increased blasts cells

2. PBS
• RBC - Normocytic & normochromic.

• WBC - Marked leucocytosis with many Lymphoblasts. Smudge cells are present.
3. Bone marrow

• Hypercellular

• Numerous blasts cells (Lymphoblasts)

4. Cytochemical staining

• Periodic Acid Shiff (PAS): glycogen in lymphoblasts

stain in magenta color.
PAS positive = ALL L1 & L2
PAS negative = ALL L3
5. Immunophenotyping
• CD (cluster determination) expression helps to identify ALL lineage based on
their surface antigens.
B-cell ALL
 Common B cell ALL
 Early pre-B ALL
 Pre-B ALL
 Mature B-cell ALL (Burkitt's leukemia)
T-cell ALL
 Pre-T ALL = CD2, CD3, CD4, CD5, CD7, CD8
 Mature T-cell ALL = CD2, CD3, CD4, CD5,CD7, CD8, CD10, CD55,CD99,…
Thank You