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Leukocytes Formation, Function and Pathology Clinical Pathology Kristin M. Canga, RVT. Leukopoiesis. All WBC production starts out in red bone marrow from the same _____________________ ______ _______ that produces RBCs.

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slide1

Leukocytes

  • Formation, Function and Pathology
  • Clinical Pathology
  • Kristin M. Canga, RVT
leukopoiesis
Leukopoiesis
  • All WBC production starts out in red bone marrow from the same _____________________ ______ _______ that produces RBCs.
  • The ____________ that act on the PPSC determine which cell type will be produced.
  • Each WBC has its own stimulus for production.
  • All WBCs differentiate and develop in the bone marrow except for some _____________________ which start out in bone marrow but develop elsewhere.
  • At the beginning of leukopoiesis, all WBCs ___________________ (in the bone marrow)
leukopoiesis1

Granulopoiesis

    • 1. Stem cell 2. Myeloblast
    • 3. Promyelocyte 4. Myelocyte
    • 5. Metamyelocyte 6. Band cell
    • 7. Mature cell (segmented neutrophil, basophilor eosinophil)
Leukopoiesis
classification of leukocytes
Classification of Leukocytes
  • Leukocytes may be classified in three different ways:
    • ____________________________
    • ____________________________
    • ______________________________________________________________________________
classification by defense function
Classification by Defense Function
  • ____________________________
    • Phagocytic leukocytes include:
      • ____________________________
      • ____________________________
      • ____________________________
      • ____________________________
  • _________________ production and cellular ______________
    • _________________________
classification by shape of nucleus
Classification by Shape of Nucleus
  • _____________________________
    • Means that the nucleus is _________________, or __________________
      • Neutrophils
      • Eosinophils
      • Basophils
  • _____________________________
    • Means that the nucleus is varying in ___________ and ______________________
      • Monocytes
  • _____________________________
    • Means that there is a _____________, rounded nucleus
      • Lymphocyte
classification by granules
Classification by Granules
  • ____________________(Presence of granules)
    • ________________– granules not attracted to stain
    • ________________– granules attracted to acidic stain
    • ________________– granules attracted to alkaline stain
  • ____________________(Absence of granules)
    • ________________
    • ________________
more on granulocytes
More on Granulocytes
  • Originally no granules present
    • PPSC  _________________, _________________, __________________,__________________, and finally the various granulocytes based on _________________ granules.
  • _____________________ produces first set of granules as cells mature
    • First set are called _______________________granules
  • Golgi Apparatus also produces granules that are attracted to stain
    • __________________ granules are added as cell matures to determine ____________________ of the cell.
functions of the specific granules
Functions of the Specific Granules
  • Specific Granules contain different substances depending on the cell:
    • Neutrophil granules contain____________________
      • Aid the cell in killing microorganisms after ___________________
    • Eosinophilgranules contain__________________________________________
      • De-granulate at site of allergic or anaphylactic reaction to reduce swelling
    • Basophil granules contain___________ and _________
      • Histamines are released to initiate ___________________ at site of reaction
      • Inflammation draws ____________________ to site
      • _________________ acts as local anticoagulant to keep blood flowing to injured/damaged area.
details of the granulocytes
Details of the Granulocytes
  • Recall that the granulocytes include:
    • _____________________
    • _____________________
    • _____________________
granulocytes neutrophils
Granulocytes - Neutrophils
  • Also called “polymorphonuclear” cells
  • Lobes of the ____________ are connected by thin ________________
  • As cell ages, the chromatin of the nucleus ______________ (squeezes together) and changes shape. (Think “________ and __________”)
  • As cell becomes ____________, the segments break apart, losing all evidence of chromatin. (Indicates dead cell)
neutrophil formation
Neutrophil Formation
  • Adult neutrophils take ~___-___ days to be produced by the bone marrow
  • Are being produced for release as circulating neutrophils migrate to __________ to fight microorganisms or die of old age.
  • If high ___________ for neutrophils occurs, they can be produced in less time.
neutrophil characteristics
Neutrophil Characteristics
  • Granules are considered to be ________ because they stain neither red, nor blue
  • Often identified on blood smear based on shape of ______________.
  • PMN adult cells may contain between 2-4 lobes connected by thin filaments of _____________.
    • Sometimes the chromatin is so thin that segments look separated.
    • (your book says 2-5 lobes, either is fine.)
neutrophil function
Neutrophil Function
  • Neutrophils only spend ~____ hours in circulation before migrating to __________.
  • Once in the tissue, neutrophils do not ________________ circulation
  • For this reason, the entire population of circulating neutrophils needs to be replaced ~________ times daily.
  • If demand is so high that bone marrow cannot supply enough adults, ________ cells will be released
  • If demand is still too high, progressively _______________ cells will be released.
neutrophil function1
Neutrophil Function
  • First line of defense in the blood
  • Major function is __________________ of invaders
  • Can respond to foreign microorganisms very quickly.
  • Use the blood as _________________, squeezing through endothelium cells via ______________. (PLEASE review Figure 9-5 on pg 234 of A&P book)
neutrophil function2
Neutrophil Function
  • Attracted to site of infection by ______________
    • Chemotaxis is the chemical reaction created between the microorganisms and the tissues that ____________cells.
  • __________________ cells with granules that contain __________________.
    • Lysosomes contain _______________ enzymes capable of destroying bacteria and viruses upon phagocytosis.
    • Membrane flows around invader and encases it in vacuole that is created, allowing lysosomes to digest invader.
    • ____________ is produced through increased _____ absorption, and is toxic to invaders AND ______________.
neutrophil function3
Neutrophil Function
  • Normally found in tissues that are constantly susceptible to invasion by microorganisms such as the________ and _____________tract.
  • Dead or abnormal neutrophils are disposed of by tissue __________________.
normal neutrophil count in blood
Normal Neutrophil Count in Blood

The neutrophil count in peripheral blood is kept within a specific range

(dogs: 3,000 – 11,400/µL; cats: 2,500 – 12,500/µL) in healthy animals and is controlled by three factors:

1) Release of mature neutrophils from the ______________ pool in the bone marrow into ________________________blood.

2) Rate of ______________ from peripheral blood into tissue.

3) Entrance of increase numbers of __________ into the neutrophil _________________ line.

maintaining normal neutrophil pools
Maintaining Normal Neutrophil Pools

1) Release of mature neutrophils from the ____________ pool in the bone marrow into the ________________ blood.

  • ________________ supply of mature neutrophils are “on call” and ready for __________________ release.
  • Caused by sudden __________________ movement of neutrophils into _____________.
maintaining normal neutrophil pools1
Maintaining Normal Neutrophil Pools

2) Rate of ___________ from peripheral blood into ____________.

  • Total neutrophil population in peripheral blood can enter the ___________ within a couple of hours
  • Triggered by massive acute _____________
maintaining normal neutrophil pools2
Maintaining Normal Neutrophil Pools

3) Entrance of increased numbers of _________ into the neutrophil ______________ line.

  • _________ method of control; it takes 3 to 6 days for neutrophils to mature for release
intravascular pools of neutrophils
Intravascular Pools of Neutrophils
  • The______________________________(CNP)
    • Refers to the blood contained in the ____________________________.
    • Blood samples obtained for _______________ analysis contain neutrophils from this pool.
    • The normal range for neutrophils is based on those contained in this pool.
  • The_____________________________ (MNP)
    • Composed of neutrophils that line the _________ of small blood vessels (not _______________________).
    • These neutrophils are not contained in blood sampled for laboratory analysis.
neutrophilia
Neutrophilia
  • Defined: An ______________ in neutrophils in blood circulation
  • If overall neutrophils increase, total ________ count increases causing __________________
    • To meet increased demand for neutrophils in tissue, the bone marrow releases its reserve stores of mature, and if necessary, _______________ neutrophils into the blood.
    • If a blood sample is drawn while these neutrophils are in transit, a ___________ than normal number of neutrophils will be included in the sample (__________________)
neutrophilia and stress
Neutrophilia and Stress
  • _____________________ leukocytosis:
    • Caused by exercise, ___________, or ________________
    • __________________ release results in a shift of neutrophils from the MNP to the CNP (_______________________)
    • No left shift occurs (no increase in _______ cells)
    • Count usually not more than ________ the normal value
    • Should return to normal within ___ minutes of removal of stimulus
    • Most common in_________ and animals <____mo.
neutrophilia and stress1
Neutrophilia and Stress
  • ____________________________
  • _____________________ -induced
    • Treatment with exogenous steroids
    • Steroids inhibit ______________
    • Endogenous steroids released in response to major _____________ illnesses, ________________ disturbances, and __________
    • Neutrophil count nearly _____________ – mature neutrophilia
    • No ____________/ Shift from MNP to CNP
neutrophis and stress
Neutrophis and Stress
  • Stress Leukogram – Continued
    • Expect to see_________________ , and _______________________.
    • Expect to see _________________ due to lysis of cells.
    • Expect to see ____________________.
    • Possible see ___________________ in dogs
    • Develops over several ________ and may last for several _________.
neutrophilia1
Neutrophilia
  • Other causes of neutrophilia include:
    • ____________________ – mild inflammation yields a leukocyte response similar to __________
      • severe inflammation yields neutrophilia with a _______________.
    • Bacterial infections
    • Conditions associated with extensive ________damage:
      • Burns, _______________, trauma, extensive ________, neoplasia
    • Extreme leukocytosis (with neutrophilia) may be associated with_____________________ that produce colony-stimulating factors:
      • Hepatozooncanisinfections, leukemias, and closed cavity infections (i.e. _________________, _______________)
leukemia
Leukemia
  • Literally means “_______________ blood”
  • Caused by malignant proliferation of one of the WBC types
    • In response to some unknown stimulus, stem cells in bone marrow start producing _______________ cells in one cell line at an __________________ rate.
    • Abnormal cells show up in blood and bone marrow in large numbers, usually before they are _____________ and cause a dramatic _____________in total WBC count.
    • Leukemias are considered a form of ____________ and can be ___________ or _____________.
    • Classified by type of _________ involved
    • Leukemia and leukocytosis may resemble one another; sometimes the distinction is difficult.
left shifts
Left shifts
  • _______________________ Left Shift - Neutrophilia with some ________ cells present; however, mature, _____________ neutrophils predominate.
  • ________________________ Left Shift – Neutropenia where ____________ neutrophils outnumber _____________neutrophils
    • Usually result of extreme migration of cells into tissues and/or detrimental effects of toxins.

Degenerative Left Shift

left shifts1
Left Shifts
  • Characterized by an increase in _________ (immature) neutrophils in the blood.
    • Note: It is normal for 0-300/µL band cells to be present in the blood of a healthy canine/feline.
  • Usually associated with _________________ conditions
  • Demand for neutrophils is ______than bone marrow pool.
  • Left shifts vary from___________ (slightly increased number of bands) to_____________ (metamyelocytes, myelocytes, and rarely- even promyelocytes present in blood)
  • ___________ cytoplasm often present during left shifts
toxic neutrophils
Toxic Neutrophils

Normal Neutrophils

Toxic Neutrophils

toxic neutrophils1
Toxic Neutrophils
  • Dohle Bodies may be noted in cytoplasm of toxic neutrophils.
  • Dohle Bodies are __________ and appear similar to _____________ seen in monocytes
right shift
Right shift
  • ___________________________ – refers to the presence of ___ or more distinct nuclear lobes within neutrophils.
  • A right shift reflects prolonged ___________time of neutrophils in blood and can occur as a result of:
    • Resolving chronic ___________________
    • Glucocorticoid administration
    • Hyperadrenocorticism (Cushing’s disease)
    • ___________________________________ disorders
  • May develop_____________ when blood film preparation is delayed for more than a few hours.
neutropenia
Neutropenia
  • Neutropenia will cause the total ________ count to decrease (_____________________)
  • If an infection is out of control, all the reserves of ____________________ can be used up faster than the bone marrow can replace them.
  • Such a condition signifies that the body is _____________ against the invading microorganisms.
  • Prognosis is ________ for a critically ill animal that has __________________ and ___________________________
neutropenia1
Neutropenia
  • Neutropenia can occur due to:
    • _______________________ of neutrophils (pseudoneutropenia)
    • Excessive ___________ demand or destruction of neutrophils
      • Destruction can be _______________-mediated
    • Reduced or ineffective granulopoiesis
  • Conditions that cause neutropenia:
    • Overwhelming _____________ infections (ex: septicemia)
    • Idiosyncratic drug reactions may result in neutropenia or pancytopenia (e.g. penicillins, cephalosporins, sulfonamides in dogs and chloramphenicol in cats)
    • Feline ______________ Virus
    • Canine ____________ Neutropenia(a.k.a. Gray Collie Syndrome)
eosinophils
Eosinophils
  • Formation: It takes ___ to ___ days to produce an eosinophil from a PPSC.
  • Population in circulation ~__-__% of total count
  • _______________ and ____________ pools are also found in peripheral blood (like __________________)
    • Bone marrow contains good supply of mature cells for release as needed.
  • Eosinophils migrate into tissue in just a few _______ where they spend the rest of their lives and undergo the same aging process as neutrophils
  • Granules’ color and size vary depending on the ____________.
eosinophil granules
Eosinophil Granules

Feline

Canine

Equine

Bovine

eosinophils1
Eosinophils
  • Eosinophils are attracted to, and inhibit local ____________ and hypersensitivity-related reactions.
    • Their granules contain anti-________________ substances that are released at the site of the allergic reaction.
  • They ingest substances associated with the _______________ immune response (antigen-antibody reaction complexes)
  • They have some ________________ and bactericidal functions
    • They are especially effective in phagocytosis of ______ pathogenic organisms, such as ____________, and some parasitic worms but are not protective against most _______________ infections.
eosinophils2
Eosinophils
  • Normal Eosinophil Values:
    • dogs: 100-750/µL; cats: 0 – 750/µL)
  • ___________________ is usually a result of:
    • ___________________: Skin, respiratory, GI tract
    • ___________________________: Anaphylaxis, Fleas, food, grasses, or non-specific allergens. Feline asthma, Chronic ______________ (skin, GI, respiratory, urogenital), Tumor associated (fibrosarcoma, mast cell tumor, lymphoma, etc.)
  • ____________________is difficult to detect and evaluate because their numbers are normally ________.
slide45

Feline eosinophilic leukemia. Two neutrophils (arrows) are adjacent to several eosinophils at various stages of development that include segmented, band, metamyelocyte, and myelocyte forms

basophils
Basophils
  • Formation:
    • Basophilsare produced in the bone marrow from the same PPSCs as other cells
  • Characteristics:
    • The granules of basophils are ________ soluble and often washed out during the staining procedure
    • _____________ are not always visible on a stained smear.
  • Function:
    • Basophilsare the_________ phagocyticof the granulocytes.
    • Their granules contain ____________ and _____________ which are responsible for at least part of their function (not much is known about basophil production or function).
basophils1
Basophils

Above: feline basophil (left); canine basophil (right)

basophil function
Basophil Function
  • Histamine and heparin granules
    • _______________ helps initiate __________________ and acute ___________ reactions.
    • _____________________ are attracted to the site of an allergic reaction by____________________________________released from the granules.
    • _____________ acts as a localized _____________________ to keep blood flowing to an injured or damaged area.
basophilia and basopenia
Basophilia and Basopenia

Normal Basophil Values: ________ in both dogs and cats

  • ___________________ can be associated with an __________ or _________________ reaction in the tissue.
    • Sometimes ______________ and _______________________ are seen at the same time.
  • _______________________: is not clinically significant. Basophils should make up less than ___% of all WBCs in peripheral blood.
agranulocytes
Agranulocytes
  • Recall that agranulocytes include
    • _________________________
    • _________________________
details of agranulocytes
Details of Agranulocytes
  • Monocytes are formed in the bone marrow from the PPSC population.

(Maturation: 1. ______________– 2. __________________– 3. ______________)

  • Lymphocytes originate from the PPSC in the bone marrow; however some travel to ________________ organs to develop and mature before settling into their permanent home in ________________ lymphoid tissue. (More on these later.)
monocytes
Monocytes
  • Formation:
    • Mature much ____________ and stay in the blood _____________ than neutrophils.
    • Total development time is ___-____ hours then they stay in the blood for ___-____hoursbefore entering tissue where they carry out their function.
  • Description
    • Monocytes are slightly __________than a neutrophil
    • Monocytes have a _______-_______cytoplasm, ______________nucleus, and a “_________” chromatin pattern.
monocytes1
Monocytes
  • Function:
    • Major _________________cells; are known as tissue ________________________once in the tissues.
    • Monocytes in the blood are considered _______________ tissue macrophages and are less effective ________________ than they are in the tissue.
monocytes2
Monocytes
  • Monocytes enter tissue by the process of_________________ in response to tissue damage caused by trauma or invading microorganisms.
  • _________________ respond more quickly to tissue damage, but ________________ stay around longer once they reach the damaged site and have become ________________.
  • Monocytes can also function in circulating blood to ___________________ damaged blood cells or microorganisms found in the blood (septicemia).
tissue macrophages
Tissue Macrophages
  • _____________ than monocytes in bloodstream
  • Are most prevalent in “__________” organs such as the_________, spleen, lungs, and____________________________.
  • Some tissue macrophages are free and wander through tissue, while others become ___________ in specific tissues and remain there for the rest of their life span (e.g.____________________ cells of liver).
  • Macrophages are often associated with_________________________ since they have a longer life span than neutrophils.
  • Collectively, the tissue macrophages and monocytes are known as the____________________________ phagocyte system.
the mononuclear phagocyte system
The Mononuclear Phagocyte System
  • Functions:
    • Clean up __________ debristhat remains after _________________ or infection is ______________
    • ____________ processing cells. The MPS can ingest antigens and present them on their cell membranes to the lymphocytes, which will destroy them.
    • Have ability to form multinucleated __________cellsin the tissue in response to foreign bodies (ex: granulomas).
    • Ingest foreign substances.
      • Capable of_________________and _________________. Can engulf structures beyond the phagocytic capacity of neutrophils such as fungi, protozoa, viruses, and dead neutrophils.
    • Are a major source of colony stimulating factors, _______________ that regulate inflammatory responses.
slide59

Fine needle aspirate of a granulomatous skin lesion. Neutrophils are adjacent to large macrophages that have abundant vacuolated cytoplasm and round to oval nuclei. Macrophages are derived from blood monocytes

monocytosis monocytopenia
Monocytosis & Monocytopenia
  • Normal Monocyte Values: (dogs: 150-1,350/µL; cats: 0-850/µL)
  • ____________________ is often associated with:
    • Chronic inflammatory conditions (particularly mycotic and other granulomatous infections)
    • ________________________
    • Bacteriemia
    • Corticosteroid or ____________ responses (especially in dogs)
  • _________________________is occasionally seen but usually has no __________________ significance.
lymphocytes
Lymphocytes

…To be continued.