heme lecture exam 3

Dendritic cell

lymph node macrophage

monocyte CD markers

CD11b and CD14

lymphoblast

1 or 2 blue nuclei, less basophilic than other blasts (more blue cytoplasm), lacy chromatin

leukocytosis steps

1. demarganilization (increase neutrophils by releasing from marginating pool)
2. BM release (release from storage pool)
3. Increased production (if infection persists, using mitotic pool)
4. recover (decrease neutrophils)

Alder-Reilly Anomoly

Auto-recessive
-Incomplete mucopolysaccharide degradation
-Purple granule with vacuole behind- looks like halo
-NORMAL function

Myeloperoxidase deficiency

Auto-recessive
-absence of MPO, but neuts can use alternative routes to kill so no increase in infection

Gaucher disease

Beta glucosidase deficiency, can't break down glucocerebroside, leading to lipid accumulation in macrophages
-Characterized by WRINKLED TISSUE PAPER LOOK in cytoplasm
-leukemia, splenectomy, and anemias

Neimann-Pick disease

Sphingomyelinase deficiency, can't break down lipids
-often fatal by 3 yrs old,
-enlarged livers and spleen, also related to leukemia
-Characterized by FOAMY macrophages

Sea blue histiocytes

Sea blue macrophages in liver, spleen, and bone marrow
-seen in lipid metabolism disorders

Measuring reticulocytes on Cloulter

Supervital staining and measures volume, conductivity, and scatter (VCS)

Electronic impedance

low, direct voltage measuring resistance of cell, most common

Coulter and Sysmex measuring WBC, RBC, and PLT

Impedance (electrical opposition)

Abott Cell-dyn measuring WBC, RBC, and PLT

Optical scatter and impedence

Coulter, Abott Cell-Dyn, and Siemens ADVIA measuring HGB

Modified cyanomethemoglobin (all at different nm)

Sysmex measuring HGB

SLB-Hb

Abott Diffs

MAPSS
(Multi-Angle Polymerized Scatter Separation)

RBC baby 1

Pronormoblast/ rubriblast/ proerythroblast
-80% nucleus
-Needs EPO
-deep blue cytoplasm and loose nucleus
-1% BM

RBC baby 2

Basophilic normoblast/ prorubicyte/ basophilic erythroblast
- 70% nucleus
-Beginning on HGB synthesis
-nucleus clumps begin
-basophilic cytoplasm
- 1-4% BM

RBC baby 3

Rubricyte/ Polychromatic normoblast/ polychromatic erythroblast
- 60% nucleus
- nucleus is almost completely clumped
-gray cytoplasm
- 3-5% BM

Baby RBC 4

Metarubicyte/ Orthochromic normoblast/ Orthochromic Erythroblast
- 45% nucleus
- very tight nucleus
- orangey cytoplasm
- 1-5% BM

Baby RBC 5

Reticulocyte/ polychromic erythrocyte
- no nucleus
-purple cytoplasm
- 0.5-2.5% BM

Pappenheimer bodies (RBC inclusion)

-iron inclusion
-splenectomy, hemolytic anemia

Ecchinocyte (Burr)

- Uremia (kidney disease), pyruvate kinase deficiency

Spherocytes

hereditary spherocytosis

Tear drop cell (dacryocyte)

primary myelofibrosis

Stomatocyte

Hereditary stomatocytosis, alcoholism, liver disease
- among us is an alcoholic

ALDER-REILLY ANOMALY

autosomal recessive, large purple granules in all lymphocytes
-incompletely degraded mucopolysaccharides that accumulate in lysosomes
-cells function normally

Chronic granulomatous disease

X-linked mutation affecting NADPH oxidase causing dysfunctioning neutrophils and oxidative burst
-IgG increased
-tested through Nitroblue tetrazolium test (NBT) and flow cytometry
-Cannot kill catalase-positive bacteria leading to large masses

Nitroblue tetrazolium test (NBT)

Reduced to a blue insoluble formazanpigment by O2 generated by activatednormal phagocytes (qualitative)

MYELOPEROXIDASE DEFICIENCY

Defective bactericidal activity• Benign autosomal recessive disorder
- Absence of MPO
- usually no increase in infection (use alt system)

B cell CD markers

CD19, CD20, CD21, CD22
- B cells

T cell CD markers

CD4- helper T cell (mostly in lymph nodes, 60-80% of pb T cells)

CD8- Cytotoxic T cells in BM (~35% of pb)
CD7, CD2, CD5, CD3

monoblast

blue, grey cytoplasm, round nucleus, agranular

promonocyte

irregular nucleus indented nucleus, coarser look, may have vacuoles or azurophilic granules

Philadelphia chromosome

mutation where chromosomes 9 and 22 are spliced together leading to uncontrolled tyrosine kinase activity = chronic myeloid leukemia (CML)

Pelger Huet vs. pseudo Pelger Huet

pelger huet= genetic bilobed nuclei with no implications
pseudo= CML

Chediak-Higashi syndrome

Autosomal recessive
- neutropenia and thrombocytopenia with giant lysosomes
-leads to albinism and high mortality for children

L.E. cells

* L for lupus
-seen in other autoimmune diseases
-Cells with the nuclei of other cells phagocytized inside
-Usually in CSF
- Allowed for 1st "anti-nuclear antibody test" to detect autoimmune disorders

Toxoplasmosis (toxoplasma gondii)

Parasitic infection from cat feces
-leukocytosis, lymphocytosis, negative monostat
- can cause cysts and parasite can be seen in cells

CMV (cytomegalovirus)

Leukocytosis, lymphocytosis, and reactive lymphs
- herpes virus
- usually asymptomatic or mild cold
- in breast milk
-tested with CMV or DNA testing

IgA Multiple Myeloma

-Flame cells
-More Ig than other plasma cells

Measuring Hgb

RBCs + Drabkin's reagent causes Hgb to be released to be measured at 540 nm and compared with Beer's law

Measuring reticulocytes on Sysmex

supervital stain, measures fluorescence

Measuring reticulocytes on Abbot CELL-DYN

Proprietary stain, multi angle scatter and fluorescence measured

Measuring reticulocytes on Siemans ADVIA

Supervital stain, measures high and low angle scatters and absorbance

Side scatter

90 degrees, measures internal complexity through refraction of internal structures, granules, and nuclear lobes

Forward scatter

0 degrees, measures size through diffraction of cell

Radio frequency

Alternating current resistence
-sometimes used alongside electronic impedence

7 degree scatter

measures complexity on 5 part differential

Siemens ADVIA measuring WBC, RBC, PLT

Optical scatter

Sysmex diffs

optical scatter and fluerescent staining

Siemens diffs

Peroxidase staining, optical scatter, and absorption for all except
basophils: lyse, laser, high and low scatter used

3 part differential

Granulocytes (not neut), Lymphocyte & Monocytes
-Sysmex
(forward scatter on x axis and side scatter on y axis)

5 part differential

Top to bottom, progressively going left as it goes down
Basophils
Neutrophiles
Monocytes on left and eos on right
Lymphocytes
nRBCs

size on y axis (0 degrees) and complexity on x axis (7 degrees)

-COULTER, Abbot Saphire, Mindray

QBC Star

single tube centrifuge and read station
- directyly measures grans, lymphs/monos, Hct
- derives Hgb
-calculates MCHC

-uses dye to differentiate the components

Basophilic stippling (RBC inclusion)

-RNA inclusion
lead intoxication

Howell-Jolly bodies (RBC inclusion)

-DNA inclusion
splenectomy, anemias

Cabot ring (RBC inclusion)

- chemo and myelodyplastic syndrome

Acanthocyte (Burr cell)

severe liver disease and splenectomy

Schistocyte

Clot in vessels, anemias, severe burns

Sickle (drepanocyte)

homozygous hemoglobin S disease (sickle cell anemia)

Elliptocyte/ ovalocyte

elliptocyte = cigar shape
ovalocyte= egg

hereditary ellipocytosis or ovalocytosis

dohle bodies

associated with burns, infections, and cancer
-composed of rough ER RNA

toxic granulation

associated with burns, infections, and cancer
-primary granules

cytoplasmic vacuoles

associated with burns, infections, and cancer

fungi inclusion

slightly larger than bacteria, round or oval
-often in immunosuppressed patient

Morulae

basophilic, irregularly shaped granular, cytoplasmic inclusions found in leukocytes in an infectious disease called ehrlichiosis.

Pelger-Huet

Autosomal dominant condition with bilobed nuclei
-no known adverse effects
-need to know so don't think theres left shift

Acquired/ psuedo pelger huet

myeloproliferative state, hypo granular, intense clumping, round nucleus

CHÉDIAK-HIGASHI SYNDROM

Rare, recessive disorder
-neutropenia, thrombocytopenia
-albinism
-fusion of neutrophil primary and secondary granules, issues with chemotaxis

May-Hegglin Anomaly

autosomal dominant trait disorder where neutrophils have blue-staining inclusions that resemble Dohle bodies; thrombocytopenia is also present with giant abnormal platelets

Dihydrorhodamine Test (DHR test)

Normal cells will reduce DHR to fluorescence

Leukocyte adhesion deficiency: LAD Type 1

Rare autosomal recessive disorder•
↓ or absent leukocyte cell-surface adhesion proteins(CD11/CD18 complex -> β2 integrins)• Unable to recognize C3bi complexes• So No phagocytosis of microbes

• Diagnosis—flow cytometry for CD11b/CD18• Treatment—prophylatic antibiotics105

NK markers

CD56 and CD16

Alveolar mac

lung macrophage

kupffer

liver macrophage

microglia

brain macrophage

osteoclasts

bone macrophage

langerhans

skin macrophage

splenic macrophage

spleen macrophage

macrophage CD markers

CD11b and CD14
GAIN CD68

monocyte

big cell, foldy nucleus, ground glass appearance, blue-grey cytoplasm, vacuoles

macrophage

huge, weird shape, round nucleus, vacuoles, in tissue but not PB
-nurse rbc in bm

Prolymphoblast

slightly smaller than normal lymphocyte, nuclei still present, clumpy chromatin

marginating pool

neutrophils on blood vessel walls

circulating pool

freely moving neutrophils in blood

mitotic pool

bm cells actively making neutrophils

storage pool

mature neutrophils in bm ready to go

LAP score

Low =possibly CML
High = prolly leukemoid reaction

Leukocyte-adhesion deficiency (LAD)

Auto-recessive
-No surface adhesion proteins on leukocytes (CD11/CD18 -> Beta integrin)
-NO PHAGOCYTOSIS
-tested with flow cytometry

EBV (mono)

leukocytosis, lymphocytosis, elevated CRV (c reactive protein)
- heterophiles antibodies and blasts present
- reactive lymphocytes and increased liver enzymes

Coulter DIffs

VCS
(volume, Conductivity, Scatter)