Lab Evaluation of WBC Disorders (Final)

In acute leukemia, a differential will show _______ cells.

blasts/immature

In acute leukemia, the patient's CBC RBCs will show _______ anemia, and PLTs will be ______.

normocytic normochromic, markedly decreased

In acute leukemia, the WBC count is ______.

variable (low-->high)

In chronic leukemia, a differential will show mostly ______ cells.

mature

Chronic leukemia will also show ______ anemia and WBC count is usually ______.

normocytic normochromic, markedly elevated

With chronic leukemia, PLT count is _______.

variable (depends on condition)

What are some diagnostic tests for WBC disorders?

- Bone marrow aspirate and biopsy

- Special stains

- Cytogenetics

- Immunophenotyping/Flow Cytometry

- Molecular diagnostics

Classifications of WBC disorders shifted from the ______ in the 1970-80s to the ______ more recently.

FAB, WHO/ICC

WHO and the ICC base classification off of ...

- Cell lineage

- Dominant clinical and biologic attributes

- Flow cytometry, cytogenetics and molecular detection of mutations

The purpose of cytochemical stains is to enhance ______ in leukocytes to help diagnose a disease or its prognosis.

identifiable substances

Cytochemistry is the study of ______ elements inside cells.

chemical

Cytochemistry can be ______ or ______.

enzymatic (peroxidase), non-enzymatic (lipids and glycogen)

Cytochemical stains can be used on what types of samples?

- Blood

- Bone marrow

- Spleen

- Lymph nodes

For acute leukemias, what stains are used?

- Myeloperoxidase (MPO)

- Esterases (NSE and SE)

Also PAS and SBB (not so much)

What cytochemical stains are used for chronic leukemias?

- Leukocyte Alkaline Phosphatase (LAP)

- Tartrate-resistant Acid Phosphatase (TRAP)

MPO is present in the ______ granules of what cells?

primary

Granulocytes and monocytes

Do lymphocytes have MPO?

No

MPO must be in ______ to be clinically significant.

blasts

______ are strongly positive with MPO.

Auer rods

Blasts of AML will be ______ with MPO.

positive

A positive MPO will be ______ in color.

black

ALL will be ______ with MPO.

negative

NSE = ______

alpha naphthyl butyrate

NSE is positive in ______ leukemias and negative in ______ leukemias.

monocytic, granulocytic

A positive NSE will be ______ in color.

orange

SE = ______

naphthol AS-D chloroacetate

Chloroacetate esterase is present in primary granules of ______.

neutrophils

SE is positive in the ______ cell line and negative in the ______ cell line.

myeloid, monocytic

A positive SE will be ______ in color.

blue

Cytogenetics can be used to ______.

diagnose diseases

The short arm of the chromosome is designated with a ______ and the long arm with a ______.

p, q

Nomenclature for chromosomes

1. # of chromosomes

2. Sex chromosomes

3. Abnormality (if present)

Example of normal chromosome nomenclature

46, XX (female)

46, XY (male)

What are the types of numeric chromosomal abnormalities?

Aneuploidy and polyploidy

Aneuploidies can be ______ or ______.

trisomies, monosomies

Trisomy occurs when a person has one ______ chromosome, and monosomy occurs when a person has one ______ chromosome.

extra, missing

Polyploidy occurs when ______ have an extra chromosome.

all chromosomes

Chromosomes can also undergo ______ in structure.

breakages

These can occur with subsequent ______ in an abnormal combination.

rejoining

Nomenclature for a male with trisomy on chromosome 8

47, XY, +8

Nomenclature for 20 cells examined, trisomy 8 in 10 cells, the rest are normal

47, XY, +8[10]/46, XY[10]

Nomenclature example for polyploidy

69, XXY

Translocation Nomenclature

1. t

2. Lowest chromosome number

3. Origin band number

4. Highest chromosome number

5. Destination band number

Deletion Nomenclature

1. del

2. Same as translocations

Nomenclature for a translocation between short arm of chromosome 12 at band p13 and the long arm of chromosome 21 at band q22

t(12;21)(p13;q22)

Nomenclature for a deletion of the long arm of chromosome 5 at band 31

del(5)(q31)

Leukomogenesis

Step-wise process of multiple mutations that affect cellular pathways and proliferation of clone

Mechanism #1 of Oncogenesis:

Uncontrolled ______ caused by mutations in genes coding for signal transduction proteins or growth factor receptors.

proliferation

This causes ______ without stimulus.

activation

Mechanism #2 of Oncogenesis:

Loss of DNA ______ capability and ______ control.

repair, cell cycle

Mechanism #3 of Oncogenesis:

Mutations in genes coding for nuclear ______ or ______ regulation.

transcription factors, epigenetic

This blocks ______.

differentiation

Mechanism #4 of Oncogenesis:

Inhibition of ______.

apoptosis

Proto-oncogene mutations are ______ of function.

gain

Tumor suppressor genes normally turn cell division ______ (on/off).

off

Tumor suppressor gene mutations are ______ of function, so cell division is always on.

loss

Lab evaluation can be used in detection of chromosomal _______ in hematological malignancies.

translocations

What are some of these hematological malignancies?

- CML

- Myelodysplastic Syndrome

- CLL

- Non-Hodgkin Lymphoma

Chronic Myelogenous Leukemia (CML) translocation nomenclature

t(9;22)

Myelodysplastic Syndrome has ______ translocations.

various

Hairy Cell Leukemia (CLL) translocations occur in ______ and ______.

IGHV, BRAF V600E

Non-Hodgkin Lymphoma translocations occur in ______ and ______.

Ig, TCR

Lab evaluation is also essential in identifying ______ hematological disorders.

inherited

What are some of these inherited hematological disorders?

- Hemoglobinopathies

- Thalassemias

- Erythrocyte disorders

In addition, identification of hematologically important ______ is important.

infectious diseases

These include ...

- Malaria

- Babesia

- Trypanosoma

- CMV

- EBV

Lab evaluation can also be important in monitoring ______ after cancer treatment.

minimal residual disease (MRD)

Molecular oncology methods

- PCR

- RT-PCR to amplify mRNA

- DNA sequencing by capillary electrophoresis

- NGS

- Chromosomal microarrays

Flow cytometry measures ______ properties of cells.

physical

This is based on their ability to ______.

deflect light

Flow cytometry characterizes the ______ properties of an unknown cell.

immunophenotyping

Immature B cell Markers

TdT, CD10, 34

Mature B cell Markers

CD19, 20, 22

Immature T cell Markers

CD34, 45, TdT, HLA-DR

Mature T cell Markers

CD2, 3, and CD4 or 8

Myeloid AML with Minimal Dif Markers

CD13, 33, HLA-DR

Myeloid APL Markers

CD13, 33, lack of HLA-DR

Monocytic AMML and AMoL Markers

CD13, 14, HLA-DR