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WBC practical
WBC practical
75
Medical
Undergraduate 4
04/30/2010

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Term
What cells are stained by myeloperoxidase stain and how do they stain?
Definition
MYELOID cells stain primary granules a dark brown-black precipitate...also LIGHTLY stains MPO found in monocytes
Term
Purpose: myeloperoxidase stain
Definition
differentiates myeloid from lymphoid blasts..is the most sensitive and specific stain for granulocytes
Term
What cells are stained by the sudan black stain and how do they stain?
Definition
Sudan Black stains the phospholipid membrane of primary granules a BLACK color

It also lightly stains the primary granules of monocytoid cells a black color
Term
What is the purpose of the Sudan Black stain?
Definition
The results parallel the myeloperoxidase stain
Term
When would you want to use the Sudan black stain over the MPO stain?
Definition
Use sudan black over MPO when fresh specimens are not avaliable
Term
What cells stain with specific esterase? How do they stain?
Definition
Myeloid cells...they stain dark blue with naphthol AS-D chloroacetate
Term
What cells stain with non-specific esterase? How do they stain?
Definition
Monocytoid cells...they stain brown **NSE is a specific marker for early nomos and macrophages

ALSO with NSE, see brown focal positivity in ALL lymphoblasts, dot-like + pattern in T-lymphs, also + are plasma cells
Term
Purpose: Combined esterase stain
Definition
Differentiates M4 from M5

M4: Blue AND Brown blasts...
M5: All Brown Blasts; segs stain BLUE
Term
PURPOSE: Fluoiride inhibition test
Definition
addition of NaFl inhibits staining of monocytes with non-specific esterase stain..sooo a check for AML M4
Term
What cells stain with the PAS stain and how do they stain?
Definition
MATURE granulocytes stain strongly, and they increase in intensity with increased maturity

monocytes are positive

lymphoBLASTS are positive with block-like staining

Lymphocytes are positive and granular with coarse staining

megakaryocytes stain positive with varying intensity anf pattern

ABNORMAL ERYTHROBLASTS STAIN ++
Term
What is the purpose of the PAS Stain?
Definition
This stain is useful for leukemic blasts in Erythroleukemia, Acute lymphoblastic leukemia, and acute megakaryocytic leukemia
Term
What cells stain with the LAP stain?
Definition
mature neutrophils and bands are positive..activity increases with neutrophil maturity
Term
LAP stain: purpose
Definition
To differentiate between CML and a leukemoid reaction

CML=LOW score

Leukemoid rxn=HIGH score
Term
In what disease state do we see a "leukemic histus" (arrest in maturation)
Definition
AML
Term
What is the diagnostic characteristic of the BONE MARROW in AML?
Definition
It has >20% (WHO) or >30% (FAB) BLASTS!
Term
What aspect of the cell does the PAS stain?
Definition
Glycogen
Term
Describe the pathophysiology of acute leukemia
Definition
There is a malignant transformation of stem cell.

Clonal amplification of that cell occurs but cell maturation arrests at the primative, blast stage of differentiation

As the neoplastic population increases...normal hematopoiesis decreases to inevitable and life threatening cytopenias

patients have problems with INFECTIONS and BLEEDING
Term
What are the PBS findings characteristic of AML?
Definition
Normocytic/normochromic anemia

nRBCs on PBS


AUER RODS!!!!!!!!!!!!!!!!!!

WBC is variable...sometimes low, sometimes normal, sometimes high
Term
What is the DIAGNOSTIC test for AML?
Definition
BM biopsy
Term
The absence of auer rods means..
Definition
The leukemia may be either lymphoid OR myeloid
Term
AML CD markers
Definition
CD34, CD33, CD13, CD14, CD15
Term
What is AML-M1?
Definition
Acute Myeloblastic Leukemia without Maturation
Term
What is: AML-M2?
Definition
Acute Myeloblastic Leukemia WITH maturation
Term
What is AML-M3?
Definition
Acute Promyelocytic Leukemia
Term
What are the 3 common chromosome aberrations found in AML-M1?
Definition
trisomy 8, t(9;22), t(6;9)
Term
What do we see in the PBS of someone with AML-M1?
Definition
ALL Blasts!
Term
What do we see in the PBS of someone with AML-M2?
Definition
Promyelocytes and occasionally more mature cells of the neutrophilic line, in addition to blasts, are present, plus other leukocytes
Term
What is the chromosome abnormality seen in AML-M2?
Definition
t(8;21)
Term
What do we see on the PBS of someone with AML-M3?
Definition
Abnormal promyelocytes with a folded nucleus.."butterfly promyelocytes"

Also see bundles of auer rods..."faggot bundles"
Term
What translocation is specific for AML-M3? What does this translocation involve?
Definition
t(15;17) is diagnostic for AML-M3

involves teh RAR-alpha and PML genes and results in an abnormal retinoid receptor
Term
Why do patients with AML-M3 have coagulation problems?
Definition
Inside the promyelocytes is a tissue-factor-like substance.. when released it causes coagulation problems
Term
What is a treatment for patients with AML-M3?
Definition
To give these patients TONS of retinoic acid, which forces the cells into maturation
Term
What is AML-M4
Definition
Myelomonocytic leukemia
Term
What is seen on the PBS of a patient with AML-M4?
Definition
myeloblasts, promyelocytes, monoblasts, and monocytes are present

Auer rods may be present
Term
What is the diagnostic BONE MARROW criteria for AML-M4?
Definition
Monocytes make up >20% of the non-erythroid cells of the bone marrow but NOT more than 80%
Term
What is the PBS monocyte count in AML-M4?
Definition
>5.0 x 10^9/L
Term
What is AML-M5?
Definition
Monocytic Leukemia
Term
What is the diagnostic criteria for AML-M5?
Definition
monocytic cells exceed 80% of the bone marrow cells
Term
What are the chromosome abberations seen in AML-M5?
Definition
Chromosome translocations or deletions involveing the long arm of chromosome 11
Term
What is the difference between AML-M5a and AML-M5b?
Definition
AML-M5a is an undifferentiated form, and AML-M5b is a differentiated form
Term
What is AML-M6?
Definition
Erythroleukemia (Di Guglielmo's Syndrome)
Term
What is the diagnostic Bone marrow finding in AML-M6?
Definition
Abnormal nucleated erythroid cells exceed 50% of the bone marrow cell
Term
What word can you use to describe the abnormal erythroid cells in AML-M6?
Definition
megaloblastoid
Term
What are some of the unusual RBC morphologies seen in AML-M6?
Definition
Giant cell forms
Multiple lobes
Variations in the size of the nuclear lobes
Vacuoles in the cytoplasm
megaloblastic changes
Howell-Jolly bodies
IN BM: ringed sideroblasts
Term
Descrobe how the prominent cells in erythroleukemia, ALL, and AML stain with the PAS stain
Definition
Erythroleukemia: intense granular staining in abnormal RBC precursors

ALL: PAS variable. Positivity shows bright fuchsia block-like granules

AML: Usually negative, occasional faint-diffuse stain is observed
Term
What is AML-M7?
Definition
Megakaryoblastic Leukemia
Term
What see on PBS: AML-M7
Definition
Cytoplasmic extensions of megakeryoblastic blasts may resemble platelet formation
Term
What immunologic markers do the blasts of AML-M7 reacti with?
Definition
glycoprotein 2b/3a
Term
What is AML-M0?
Definition
Minimally differentiated Myelocytic leukemia
Term
What is the normal LAP range, and what range describes CML and reactive neutrophilia?
Definition
Normal LAP range: 13-130
CML: LAP <13
Reactive neutrophilia: LAP >160
Term
t(9:22)
Definition
Philadelphia Chromosome--CML
Term
t(8;14)
Definition
Burkitt's Lymphoma--ALL L3
Term
t(8;21)
Definition
AML-M2
Term
11q23 abnormalities
Definition
AML-M5
Term
t(15;17)
Definition
retinoic acid receptor alpha AML-M3
Term
t(14;18) what does it create?
Definition
Follicular lymphoma--BCL-2 and IGH
Term
t(11;14) what are these genes
Definition
mantle cell lymphoma IGH and cyclin D1
Term
t(11;18)
Definition
diffuse large B-Cell lymphoma..this translocation makes the cells resistent to H. Pylori therapy
Term
What is myeloid metaplasia?
Definition
Myeloid metaplaisa refers to the extramedullary hematopoiesis of the myeloid cell line

This often occurs secondary to
Term
What are the two stages of CIMF/PMF?
Definition
Prefibrotic stage and Fibrotic stage
Term
Where do we see the JAK-2 mutation?
Definition
The JAK-2 mutation is found in almost all cases of PV and in approximately 50% of patients with CIMF (and ET)
Term
Describe how the JAK-2 mutation relates to the pathogenesis of disease
Definition
The mutation causes JAK2 enzyme to be constituatively active causing cells to be permenently active, and proliferate without EPO signal (EPO no longer acts as regulation mechanism)
Term
Comment on how each cell line looks on a PBS of someone with CML
Definition
Mature granulocytes: predominantly monocytic cellular pattern, DYSPLASTIC cells: hyposeg neutrophils, small and wierd looking monocytes

Plateplets: thrombocytopenia...large platelets and micromegakaryocytes

RBCs: poik and iunclusions
Term
What is the component of the Non-specific esterase stain?
Definition
ASD chloroacetate
Term
What is the component of the specific esterase stain?
Definition
Alpha-Naphthyl acetate
Term
What lab findings would help differentiate CMML from CML?
Definition
1.) PBS/BM: For CMML a noticeable monocytosis and tri-lineage dysplasia will be noticed but absent in CML (no dysplasia)

2.) Philadelphia Chromosome: Absent in CMMl and present in CML

3.) Lysozyme: elevated in both serum and urine levels in CMML but not in CML
Term
What test helps differentiate AML M0 from AML M1?
Definition
The blasts in AML-M0 display negative reactions with all cytochemical stains. On the other hand, the myeloperoxidase or Sudan Black B is positive in more than 3% of the blasts of patients with AML-M1
Term
How are AML-M5a and AML-M5b differentiated?
Definition
In M5a, monoblasts account for 80% or more of all monocytic cells in the bone marrow, while in M5b, monoblasts account for <80% of the monocytic component. Such classification is important as it provides information on the aggresiveness of the disease and the response to therapeutic regimens
Term
Describe the appearance of the bone marrow in patients with AML M6
Definition
More than 50% of the bone marrow cells in a patient with AML M6 are erythroid and 30% or more of the remaining cells are blasts. Additionally, the normoblasts found in the bone marrow may be giant and multinuclear or multilobular. The normoblasts may exhibit nuclear budding and fragmentation, cytoplasmic vacuoles, Howell-Jolley bodies, ringed sideroblasts, and megaloblastic changes. Additionally, erythrophagocytosis, auer rods within the myeloblasts, hypogranular neutrophils and pseudo-Pelger-Huet anomaly in neutrophils
Term
Explain why patients with AML M3 might have abnormal coagulation test
Definition
AML-M3 is characterized by a predominance of promyelocytes. The granules of promyelocytes contain a procoagulant which is readily introduced into the bloodstream of patients with AML-M3 when their promyelocytes release their granules or are lysed
Term
What purpose does the retinoic acid receptor serve?
Definition
Helps in cellular differentiation and maturation
Term
Compare G:E ratio in CML vs. in AML
Definition
CML: 10-50:1

AML 100:1
Term
Describe blast crisis in CML
Definition
Patients in this final phase of CML have more than 30% blasts in their PBS and BM samples. The blasts frequently invade other tissues and organs outside the bone marrow. During this phase the disease transforms into an aggressive, acute leukemia (70% AML, 30% ALL)) If untreated, CML is fatal in roughly 20% of all patients each year

In BM: FIBROSIS!!!
Term
What two cytochemical stains are usually positive in ALL but negative in AML
Definition
Tdt and PAS
Term
How do we calculate the G:E ratio?
Definition
Granulocytes:
Segs, bands, metas, myelos, promyelocytes, blasts, eos and basos

NOT Lymphs, NOT Monos

Erythrocytes: ALL erythroid precursors
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