MLT1314 hemato Lecture 1 (intro to hematology)

Granulocytes

Neutrophil

Eosinophil

Basophil

Agranulocytes

Lymphocyte (B and T)

Monocyte

Hematopoeitic stem cells are capable of?

continuous self-replication and proliferation

AND they can differentiate into stem cells of lymphoid or myeloid lineage

Myeloid lineage covers

Erythrocyte

thrombocyte

granulocytes

monocytes

Lymphoid lineage covers

lymphocytes

Mast cells are commonly mistaken for basophils, what are some main differences?

Mast cells are 6-12um (baso: 5-7um)

Lifespan: months (baso: days)

Location: tissues (baso: intravascular)

Matures in: connective tissue (baso: BM)

Granules: large (baso: small and large)

Nucleus: round (baso: lobated)

Response: localizes (baso: systematic)

What are barr bodies? When do you see them?

Seen in neuts of female PT samples

Little offshoot of nucleus, tiny ball

It is a portion of the X-chromosome

Main ways to visually differentiate between the 5 WBCs

Neut: granular, purple cyto, lobe nucleus

Lymph: smaller, darker, large nucleus

Mono: large, lilac cyto, vacuole

Eosino: orange granule

Baso: purple granules

How are metas/myelos counted on a Peripheral blood smear?

Theyre counted collectively for neuts, eosinos, and basos

How are band granulocytes counted on a peripheral blood smear?

Added to mature cell counts

Band neut=part of abs neut count

What to do if blood smear is too thin (due to anemia)?

-Increase angle of spreader slide

-Increase speed of the push

What to do if blood smear is too thick due to high RBC count?

-Decrease angle of the smear

-Decrease speed of the smear

What to do if blood smear is too thick due to viscous blood?

-Decrease angle of the smear

-Dilute blood w/ saline/diluent

What to do if blood smear has aglgutination

-Prepare slide after warming blood

What to do if blood smear has grit/particle in tail?

-Dirt on spreader slide or smear slide (get new clean slides)

-Stain deposit (filter stain or make new slides)

Size of neut, eosino, baso, mono, and lymphs

Neut 9-15um

Eosnio 10-16um

Baso 10-14um

Mono 12-20um (large)

Lymph 7-12 um (small)

Lifespans of neut, eosino, baso, mono, and lymphs

Neut 6hr-few days

Eosino 8-12 days tissue (4-12hrs circulating)

Baso few hrs-few days

Mono Few hrs-few days

B lymph yrs

T lymph yrs

Why is there no such thing as a low eosino/baso/mono count?

The reference range STARTS at 0, so its naturally low

Ref range neut

2.0-7.5 (x10^9/L)

Ref range eosino

0.0-0.6

Ref range baso

0.0-0.2

WBC ref interval

4.8-10.8

RBC size

6-8um

Neutrophilia

>7.5

Neutropenia

Neut:

Low alert

Crit low

Alert < 1.0

Critical

Neut:

High alert

Crit high

Alert > 50.0

Critical >100

Causes of neutrophilia

Bacterial infection

Pregnancy

Some cancers (CML, AML)

Vasculitis

Stress response

Surgery

Malignancies

Physical conditions (heat, cold, shock)

Drugs

Inflammatory response

Stress response

Tissue damage (surgical, trauma(

Neutrophilia goes hand-in-hand with a high _______?

WBC count (leukocytosis)

Causes of neutropenia

Chemo and other drugs

Early infectious mono

Environmental exposure

Aplastic anemia

Septicemia / infectious disease

Cyclic neutropenia (intervals of decimated neuts)

What does a "left shift" mean?

Neut population is showing immaturity

Normal band range

0-0.7 (high is >0.7)

Can a left shift still be noted if

Yes, only if its accompanied by other immature cells

What does a blast cell look like?

-Possible auer rods

-N:C = 4:1

-smooth, lacy, even chromatin

-large round nucleus

WBC count is naturally higher in children, slowly lowering until 17yrs of age. T or F

TRUE

What does a "leukoerythroblastic blood film" refer to?

Granulocyte left shift

Nucleated RBCs

Very serious condition (MDS)

When do primary grnaules appear in WBCs? When do they go away?

In immature forms of granulocytes

Disappear as the cell grows from promyelo to myelo

Look azurophilic

When do secondary granules appear in WBCs?

Increase in # as cell matures

Late promyelocyte - myelocyte stage

What do you do if you count 0 neuts on a blood film?

Count another 100

IF still seeing none after counting 200 cells, scan entire readable area

Report out of 200 cells

Report as "rare neut seen on scan" or "0" is none were TRULY seen

What will you see in bacterial infection?

Leukocytosis (increase in total WBC count)

Neutrophilia (increase in ANC)

Left shift (band neut count > 0.7)

TG/TV

Dohle bodies

What consitutes a toxic change?

TV, TG, and dohle bodies

What are dohle bodies?

Light-blue smear in cyto of neuts

Theory--> remnants of rRNA or rER due to accelerated granulopoiesis

High dohle body count means?

Consider bacterial infection

Seen in May Hegglin anomaly

What is toxic granulation (TG)?

Larger/darker granules than normal

Composed of peroxidase

Why does toxic vacuolization (TV) occur?

Results from phagocytosis and digestion of foreign particles w/in the cytoplasm

In what cells can TG be seen?

Metamyelocytes, Band Neutrophils and mature Neutrophils

Phagocytosis is carried out by what cells?

Neuts and monos

4 steps of phagocytosis

Chemotaxis

Opsonization

Ingestion

Killing

Chemotaxis

Chemical signals that trigger the neuts to respond

Opsonization

Preparation of the foreign body for attachment by the use of complement and immunoglobulins

Ingestion (phago)

cell membrane of the neut/monocyte engulfs/wraps itself around the invading body until it has completely engulfed the body within the newly formed vacuole = phagosome

Killing (phago)

The lytic or digestive elements contained within the lysosome granules are released, destroying the particle and the neutrophil itself

Right shift means?

Decrease in # of immature neuts + increase in # of mature neuts

When do you see a right shift?

When recovering from infection or inflammation

When are crystals of death seen ("green cytoplasmic inclusions") in neuts and sometimes monos

Advanced liver failure

Multiple organ failure

When seeing hypo/agranular neuts, consider?

myelodysplastic syndrome

B12/folate deficiency

Hypersegmented neuts have how many lobes?

Several neuts w/ 5 lobes OR

1 neut w/ 6+ lobes

Hypersegmented neuts are sometimes seen w/ shift to the left. T or F

FALSE. Seen w/ shift to the right

Also can be seen as a hereditary conditon

Name 4 hereditary neut disorders

Pelger huet anomaly (PHA)

May hegglin anomaly (MYH9 disorders)

Alder reily anomaly (ARA)

Chediak-Higashi anomaly

PHA lobes vs pseudo-PHA lobes

PHA --> bilobed neuts w/ fine filament connection

Pseudo-PHA--> pince-nez w/ ~

Harlequin cells are aka?

Dysplastic eosinophil

When are harlequin cells seen

Common: CML

Sometimes: myelodysplastic syndrome (MDS)

What is a harlequin cell?

Eosinophils with atypical basophilic staining granules.

the cell couldnt decide wtf it wanted ot be

Eosinophilia

>0.6

Eosinophilia causes

Allergies

Asthma

Skin disease (eczema, psoriasis, etc.)

Parasitic infections

Myeloproliferative disorders

Basophilia

>0.2

Basophilia common causes

Malignancies

Myeloproliferative disorders (esp CML)

Allergies, asthma, anaphalaxis

Infection/autoimmune disorders

Chicken pox

small poc

Influenza

Tuberculosis

Basophilia rarer causes

Ulcerative colitis

Chronic inflammatory conditions (RA)

Parasitic infection

In terms of WBCs, how would a bacterial infection present

Usually leukocytosis neutrophilia

May have: left shift, TG/TV, dohle bodies

In terms of WBCs, how would pregnancy present

Usually mild leukocytosis neutrophilia

May have slight left shift

No toxic change (unless infection)

In terms of WBCs, how would a leukemoid rxn present

Exaggerated/severe neutrophilia

Often WBC > 50.0x109/L

May have: left shift, TG/TV, dohle bodies

In terms of WBCs, how would malignancy present

Usually leukocytosis neutrophilia basophilia

Left shift w/ immature cells are present (esp earlier band forms)

4 kinds of neutrophil variations we keep track of

TG

TV

Hypogranulation

Hyposegmentation

(Dohle bodies, etc)

Describe the appearance of secondary granules in neuts, eosinophils, and basophils

Neut→ neutral stain (mix dye), purple

Eosino→ basic stain (acid dye), red/orange

Baso→ acidic stain (basic dye), dark blue

CML

chronic myelogenous leukemia

A wrights giemsa stain is composed of 2 dyes, name them

Eosin Y

Methylene Blue

How does eosin Y dye work

Its an acidic anionic dye (neg charge)

Stains basic components (pos charge) = eosinophilic

Name basic components of a cell that eosin Y would dye

Cytoplasm, eosino granules, RBCs

Name acidic components methylene blue/azure B would dye

Nuclear chromatin, alkaline cytoplasm, baso granules, DNA, RNA

How does Methylene blue dye work

Basic cationic dye (pos charge)

Stains acidic components (neg charge)

Eosinophils can "degranulate," what does that mean?

They release cytotoxic proteins directed at invader.... but this is also toxic to normal tissue = allergic reaction

What do eosinophil granules contain?

Granules contain enzymatic and non-enzymatic proteins to destroy targeted cells.

Eosinophils cant phagocytize. T or F

FALSE, they can

Where are eosinophils found?

Held in tissue

Released into circulation when needed

What do basophil granules contain

histamine, heparin and serotonin (all released during immune response)

Why would a basophil degranulate? And what happens when they do?

Triggered by inflammation, infection, allergy

Burst granules, release contents (histamine, herpain, serotonin)

Also release proteolytic enzymes (peroxidase)

Histamine function

increase blood flow to area

Heparin function

Anticoagulant

What is sepsis/Septicemia (aka bacteremia)

Infection in the bloodstream

How does septicemia present?

Leukocytosis

Neutrophilia

Left shift

Immature forms (metas, myelos)

TG/TV/Dohle bodies

Look for intra/extracellular organisms

Primary infection

Other BFs may have pos cultures

Septicemia is often caused by fungi/viruses. T or F

FALSE, its often bacterial

Symptoms of septicemia

Widespread inflammation

Fever, chills

Rapid heart rate

Rapid breathing

Confusion or disorientation

Low BP

Decreased urine output

Extreme fatigue

Organ failure

Septicemia can present with low WBC count. T or F

TRUE, severe sepsis MAY have leukopenia and neutropenia (this is an emergency)

Dying cells (apoptosis) look pyknotic, and this is often seen when?

Artifact as blood sample ages

PT w/ increased destruction

PT w/ splenectomy

Pyknotic

condensation of chromatin as the cell undergoes apoptosis/necrosis

BASIC inflammatory response (may be out of order a bit)

--Pathogen enters body

--Early in, neu count is low

--24hrs needed to inc neut prod

--AG triggered

--Vasodilation

--Cytokines/inflamm factors stim neut prod

--Immune cell, protein, nutrients move in

--Chemoattractants draw neuts in

--Neuts activate/do their thing

--Leukocytosis/neutrophilia occurs

--Phagocytes clean up

LAP stands for

Leukocyte alkaline phosphatase

LAP score helps us do what?

ID infection (inc WBC/leukemoid rxn)

vs

leukemia (CML)

How does LAP score help differ between infection and leukemia?

LAP stains enzymes in mature neutrophils

The azo dye forms an enzyme complex (insoluble brown precipitate)

What blood tube is used to collect for an LAP score

Peripheral blood or BM

NO EDTA

Do heparin or capillary collection