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Developed by
Dr.Abdulrazzaq Othman Alagbare-MD-MCP-
Hematology Lecturer
INTRODUCTION
hematological- malignant Disorders
FOR Clinical LABORATORY STUDENTS
The Malignant Hematological Disorders
Means malignancy for all blood cells that produce from the bone marrow
1-Leukemia for the following
cells
a) Neutrophil
b) Eosinophil
c) Basophil
d) Monocyte
e) T-Lymphocyte
f) B-Lymphocyt
2-RBC:
Polycythemia Vera (PV)
3-Thrombocytes : Essential
Thrombocythemia (ET)
4-Plasm cells
Multiple myeloma (MM)
Introduction-Leukemia
Definition: Leukemia is a cancer of blood white cells
Caused by the mutation of
pluripotent or most
primitive stem cells. (PSC,
MSC, LSC)
Blast cells accumulate in the Bone
marrow and
1. peripheral blood
2. Spleen
3. Lymph nodes
4. Liver
The leukemic cells are
1. Trapped early from the B.M
2. Proliferate without control (No effect of growth factors)
3. Not able to carry out their function
4. fetal disease if not treated
1
1- Myeloid group malignancies
Granullar cells ––> (N. E. and B ) Leukemias
Monocytes ––> Monocytic leukemia
Erythrocytes ––> Polycythemia vera
Platelets ––> Essential thrombocythemia
2- Lymphoid group malignancies
B-lymphocytes ––> Leukemias
T-lymphocytes ––> Leukemias
Plasma cells ––> Multiple myeloma
Causes of leukemia or Predisposing factors
1. Inherited factors
2. Environmental
factors
3. Infection
genetic diseases e.g.
Down's syndrome,
Klinefelter's syndrome,
Fanconi's anemia,
Chemicals e.g.
Benzene
Drugs e.g.
Alkylating agents
Radiation
Viruses e.g. Epstein-Barr virus EBV, HIV,
Human herpes virus 8 (HHV-8)
Bacteria e.g. Helicobacter Pylori
Protozoa e.g. Malaria
FAB classification
The FAB classification is based largely on.
1-Morphology of cells
2-Simple cytochemical stains
1. At least 30% of cells in the
bone marrow or blood must be
myeloblasts.
2. and does not include cytogenetic
abnormalities
But must be
Differences between acute and chronic leukemia
1- Cells maturation degree (blast or mature )
2-Blood cells count in the peripheral blood
3-Patient`s symptoms
4-Clinical onset
5-organomegaly
6-patient`s age
All Leukemia divided into Acute and chronic
Properties of acute leukemia Properties of chronic leukemia
 Immature cells (90% blast cells )  More mature cells (60%-70% mature cells)
 Occurs in all ages  Usually occurs in adults and elderly
 Clinical onset is sudden  Clinical onset is gradual
 Anemia and thrombocytopenia are severe  Anemia and thrombocytopenia are mild
 WBC is variable (high, normal or low with 90 %
blast cells)
 WBC is increased , abnormal cells, (30% blast )
 organomegaly is mild  Organomegaly is prominent
Clinical Picture of acute
leukemias
Clinical Picture of chronic
leukemias
The patient has
1. Anemia
2. Bleeding
3. Infection
The patient has
1. No anemia
2. No bleeding
3. No Infection
Acute leukemia cause morbidity and mortality through :-
1. Deficiency in blood cell number and function
2. Invasion of vital organs
3. Systemic disturbances by metabolic imbalance
Pathophysiology
Acute leukemia cause morbidity and mortality through :-
1. Deficiency in blood cell number and function
2. Invasion of vital organs
3. Systemic disturbances by metabolic imbalance
Investigations need to diagnose leukemia
1. Complete blood count.(CBC or FBC)
2. Peripheral blood film inspection
3. A bone marrow examination
4. Flow cytometry or immunophenotyping studies
5. Chromosomal analysis
6. Cytochemical stains
1-Complete blood count show
1. The RBC changes
2. WBC: count,
3. Platelets: count and morphology
PBS: show
1. Changes of white blood cells,
2. % of blast cells
3. Type of blast cells (myeloblast or lymphoblast etc)
4. morphologic feature (auer rods, vacuoles , the size of the
nucleus etc)
2-A bone marrow examination
• mostly has hypercellular B.M and few cases hypocellular
• 20% to 90% leukemic blasts at diagnosis or during relapse.
• The blast must present in the peripheral blood, unless the WBC count is markedly
decreased.
Acute leukemia
Bone Marrow Tryphine Biopsy
It is a histological test for bone marrow tissue to obtain the blood cells
Indication: If the peripheral blood indicate
1-hypocelluar
2-aplastic anemia
3-metastic cancer
Or the previous result indicate  Dry tap bone marrow aspiration
3-Flow cytometry or immunophenotypic studies
Monoclonal antibodies toward cell-type restricted antigens are used in this
highly specific method
CD markers Show for
1. blast cells
2. mature cells
and determine if it is
1. myeloid Cells types and count
2. lymphoid cells types and count
3. Normal or abnormal
4- Chromosomal analysis
Chromosomal abnormalities studies is very important (diagnostic
and prognostic ) for
• AML
• ALL
critical in the diagnosis and treatment of AML.As in
ALL
Note
1-AML: Acute myeloblastic Leukemia
2-ALL: Acute Lymphoblastic Leukemia
5-Cytochemical stains
Importance: very helpful in the diagnosis and classification of acute
leukemias
Specimens: bone marrow smears but may also be done on peripheral
smears
1. myeloperoxidase (MPO)
2. Sudan black B stain (SBB)
3. specific esterase stain (SE)
4. a non specific esterase stain (NSE)
5. Terminal deoxynucleotidyl transferase (TdT)
Types
Cytochemical Reaction Cellular Element Stained Blasts Identified
Myeloperoxidase
(MPO)
Neutrophil primary granules Myeloblasts strong positive;
Sudan Black B (SBB) Phospholipids Myeloblasts strong positive;
Specific esterase Cellular enzyme Myeloblasts strong positive;
Nonspecific esterase
(NSE)
Cellular enzyme Monoblasts strong positive
Terminal
deoxynucleotidyl
transferase (TdT)
Intranuclear enzyme Lymphoblasts positive
-Positive-Myeloperoxidase- (MPO)
Brown black deposits
Positive-Chloracetate (Specific) Esterase-Myeloid
Cell Line
Red deposit Brown deposits
Positive-Non-Specific Esterase
Monocytic Line
Leukocyte Alkaline Phosphatase (LAP, Neutrophil Alkaline
Phosphatase, NAP)
Definition: The LAP score is a test done by performing a chemical reaction
on a blood smear
Evaluation:
1. The blood smear is viewed under a microscope, and the degree of granular staining
in mature neutrophils (bands and segs) is graded from 0 (no staining) to 4+ (dense
granular staining).
2. The LAP score is the sum of staining for 100 cells.
3. The normal range is 20 to 100.
Principle: The granular of bands and segs with the alkaline phosphatase
enzyme colored staining.
LAP Score
• Count 100 consecutive segs and
bands
• Score:
0 = no granules
1+ = occasional diffuse granules
2+ = moderate number of granules
3+ = many strongly positive granules
4+ = confluent strongly positive
granules
Example:
0 x 35 cells = 0
1+ x 30 cells = 30
2+ x 20 cells = 40
3+ x 10 cells = 30
4+ x 5 cells = 20
120
LAP Score
0 1+
2+ 3+ 4+
The two main conditions with
a low LAP score are
1. CML
2. paroxysmal nocturnal hemoglobinuria (PNH).
higher scores in
1. Polycythemia vera.
2. Myelofibrosis
3. Essential thrombocytopenia
4. leukemoid reactions
LAP score and related diseases
Precuations
1. Reject specimens collected in EDTA-anticoagulated (lavender-topped) tubes because EDTA inhibits the
activity of LAP
2. Results are invalid if the client is neutropenic (that is, <1000/mm3 neutrophils).
Instrumentation and leukemia
Platelets
The platelets count increased due to fragments of the
leukemic blasts, which are counted as platelets
To solve this problem
1. Platelets must be counted manually
2. Examination of a blood smear gives a more accurate
assessment of the platelet count in this circumstance.
Instrumentation and leukemia
WBC
Must check
1-The count on the blood smear
2-the DLC, if not agree count to 200 cells ,
report the average
Be carful for
1-the clot , old . Lipemic specimen, affects the results of CBC
2-the age of the patient (infant, child, adult) must be taken into account
3- newborns, toddlers, and young children, particular reference ranges must be taken
into account
4- a normal range is given, covering 95% of the values of healthy persons
5-The presence of abnormal cells needs clinical pathologist
END OF THE
LESSON

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Introduction of Hematol.Malignancy .ppt

  • 1. Developed by Dr.Abdulrazzaq Othman Alagbare-MD-MCP- Hematology Lecturer INTRODUCTION hematological- malignant Disorders FOR Clinical LABORATORY STUDENTS
  • 2. The Malignant Hematological Disorders Means malignancy for all blood cells that produce from the bone marrow 1-Leukemia for the following cells a) Neutrophil b) Eosinophil c) Basophil d) Monocyte e) T-Lymphocyte f) B-Lymphocyt 2-RBC: Polycythemia Vera (PV) 3-Thrombocytes : Essential Thrombocythemia (ET) 4-Plasm cells Multiple myeloma (MM)
  • 3. Introduction-Leukemia Definition: Leukemia is a cancer of blood white cells Caused by the mutation of pluripotent or most primitive stem cells. (PSC, MSC, LSC) Blast cells accumulate in the Bone marrow and 1. peripheral blood 2. Spleen 3. Lymph nodes 4. Liver The leukemic cells are 1. Trapped early from the B.M 2. Proliferate without control (No effect of growth factors) 3. Not able to carry out their function 4. fetal disease if not treated 1
  • 4. 1- Myeloid group malignancies Granullar cells ––> (N. E. and B ) Leukemias Monocytes ––> Monocytic leukemia Erythrocytes ––> Polycythemia vera Platelets ––> Essential thrombocythemia 2- Lymphoid group malignancies B-lymphocytes ––> Leukemias T-lymphocytes ––> Leukemias Plasma cells ––> Multiple myeloma
  • 5. Causes of leukemia or Predisposing factors 1. Inherited factors 2. Environmental factors 3. Infection genetic diseases e.g. Down's syndrome, Klinefelter's syndrome, Fanconi's anemia, Chemicals e.g. Benzene Drugs e.g. Alkylating agents Radiation Viruses e.g. Epstein-Barr virus EBV, HIV, Human herpes virus 8 (HHV-8) Bacteria e.g. Helicobacter Pylori Protozoa e.g. Malaria
  • 6. FAB classification The FAB classification is based largely on. 1-Morphology of cells 2-Simple cytochemical stains 1. At least 30% of cells in the bone marrow or blood must be myeloblasts. 2. and does not include cytogenetic abnormalities But must be
  • 7. Differences between acute and chronic leukemia 1- Cells maturation degree (blast or mature ) 2-Blood cells count in the peripheral blood 3-Patient`s symptoms 4-Clinical onset 5-organomegaly 6-patient`s age All Leukemia divided into Acute and chronic
  • 8. Properties of acute leukemia Properties of chronic leukemia  Immature cells (90% blast cells )  More mature cells (60%-70% mature cells)  Occurs in all ages  Usually occurs in adults and elderly  Clinical onset is sudden  Clinical onset is gradual  Anemia and thrombocytopenia are severe  Anemia and thrombocytopenia are mild  WBC is variable (high, normal or low with 90 % blast cells)  WBC is increased , abnormal cells, (30% blast )  organomegaly is mild  Organomegaly is prominent
  • 9. Clinical Picture of acute leukemias Clinical Picture of chronic leukemias The patient has 1. Anemia 2. Bleeding 3. Infection The patient has 1. No anemia 2. No bleeding 3. No Infection Acute leukemia cause morbidity and mortality through :- 1. Deficiency in blood cell number and function 2. Invasion of vital organs 3. Systemic disturbances by metabolic imbalance
  • 10. Pathophysiology Acute leukemia cause morbidity and mortality through :- 1. Deficiency in blood cell number and function 2. Invasion of vital organs 3. Systemic disturbances by metabolic imbalance
  • 11. Investigations need to diagnose leukemia 1. Complete blood count.(CBC or FBC) 2. Peripheral blood film inspection 3. A bone marrow examination 4. Flow cytometry or immunophenotyping studies 5. Chromosomal analysis 6. Cytochemical stains
  • 12. 1-Complete blood count show 1. The RBC changes 2. WBC: count, 3. Platelets: count and morphology PBS: show 1. Changes of white blood cells, 2. % of blast cells 3. Type of blast cells (myeloblast or lymphoblast etc) 4. morphologic feature (auer rods, vacuoles , the size of the nucleus etc)
  • 13. 2-A bone marrow examination • mostly has hypercellular B.M and few cases hypocellular • 20% to 90% leukemic blasts at diagnosis or during relapse. • The blast must present in the peripheral blood, unless the WBC count is markedly decreased. Acute leukemia Bone Marrow Tryphine Biopsy It is a histological test for bone marrow tissue to obtain the blood cells Indication: If the peripheral blood indicate 1-hypocelluar 2-aplastic anemia 3-metastic cancer Or the previous result indicate  Dry tap bone marrow aspiration
  • 14. 3-Flow cytometry or immunophenotypic studies Monoclonal antibodies toward cell-type restricted antigens are used in this highly specific method CD markers Show for 1. blast cells 2. mature cells and determine if it is 1. myeloid Cells types and count 2. lymphoid cells types and count 3. Normal or abnormal
  • 15. 4- Chromosomal analysis Chromosomal abnormalities studies is very important (diagnostic and prognostic ) for • AML • ALL critical in the diagnosis and treatment of AML.As in ALL Note 1-AML: Acute myeloblastic Leukemia 2-ALL: Acute Lymphoblastic Leukemia
  • 16. 5-Cytochemical stains Importance: very helpful in the diagnosis and classification of acute leukemias Specimens: bone marrow smears but may also be done on peripheral smears 1. myeloperoxidase (MPO) 2. Sudan black B stain (SBB) 3. specific esterase stain (SE) 4. a non specific esterase stain (NSE) 5. Terminal deoxynucleotidyl transferase (TdT) Types
  • 17. Cytochemical Reaction Cellular Element Stained Blasts Identified Myeloperoxidase (MPO) Neutrophil primary granules Myeloblasts strong positive; Sudan Black B (SBB) Phospholipids Myeloblasts strong positive; Specific esterase Cellular enzyme Myeloblasts strong positive; Nonspecific esterase (NSE) Cellular enzyme Monoblasts strong positive Terminal deoxynucleotidyl transferase (TdT) Intranuclear enzyme Lymphoblasts positive
  • 19. Positive-Chloracetate (Specific) Esterase-Myeloid Cell Line Red deposit Brown deposits Positive-Non-Specific Esterase Monocytic Line
  • 20. Leukocyte Alkaline Phosphatase (LAP, Neutrophil Alkaline Phosphatase, NAP) Definition: The LAP score is a test done by performing a chemical reaction on a blood smear Evaluation: 1. The blood smear is viewed under a microscope, and the degree of granular staining in mature neutrophils (bands and segs) is graded from 0 (no staining) to 4+ (dense granular staining). 2. The LAP score is the sum of staining for 100 cells. 3. The normal range is 20 to 100. Principle: The granular of bands and segs with the alkaline phosphatase enzyme colored staining.
  • 21. LAP Score • Count 100 consecutive segs and bands • Score: 0 = no granules 1+ = occasional diffuse granules 2+ = moderate number of granules 3+ = many strongly positive granules 4+ = confluent strongly positive granules Example: 0 x 35 cells = 0 1+ x 30 cells = 30 2+ x 20 cells = 40 3+ x 10 cells = 30 4+ x 5 cells = 20 120 LAP Score
  • 23. The two main conditions with a low LAP score are 1. CML 2. paroxysmal nocturnal hemoglobinuria (PNH). higher scores in 1. Polycythemia vera. 2. Myelofibrosis 3. Essential thrombocytopenia 4. leukemoid reactions LAP score and related diseases Precuations 1. Reject specimens collected in EDTA-anticoagulated (lavender-topped) tubes because EDTA inhibits the activity of LAP 2. Results are invalid if the client is neutropenic (that is, <1000/mm3 neutrophils).
  • 24. Instrumentation and leukemia Platelets The platelets count increased due to fragments of the leukemic blasts, which are counted as platelets To solve this problem 1. Platelets must be counted manually 2. Examination of a blood smear gives a more accurate assessment of the platelet count in this circumstance.
  • 25. Instrumentation and leukemia WBC Must check 1-The count on the blood smear 2-the DLC, if not agree count to 200 cells , report the average Be carful for 1-the clot , old . Lipemic specimen, affects the results of CBC 2-the age of the patient (infant, child, adult) must be taken into account 3- newborns, toddlers, and young children, particular reference ranges must be taken into account 4- a normal range is given, covering 95% of the values of healthy persons 5-The presence of abnormal cells needs clinical pathologist