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Leukocytes Formation, Function and Pathology Clinical Pathology Kristin M. Canga, RVT

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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Leukocytes Formation, Function and Pathology Clinical Pathology Kristin M. Canga, RVT

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  1. Leukocytes • Formation, Function and Pathology • Clinical Pathology • Kristin M. Canga, RVT

  2. 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)

  3. Granulopoiesis • 1. Stem cell 2. Myeloblast • 3. Promyelocyte 4. Myelocyte • 5. Metamyelocyte 6. Band cell • 7. Mature cell (segmented neutrophil, basophilor eosinophil) Leukopoiesis

  4. Classification of Leukocytes • Leukocytes may be classified in three different ways: • ____________________________ • ____________________________ • ______________________________________________________________________________

  5. Classification by Defense Function • ____________________________ • Phagocytic leukocytes include: • ____________________________ • ____________________________ • ____________________________ • ____________________________ • _________________ production and cellular ______________ • _________________________

  6. 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

  7. Classification by Granules • ____________________(Presence of granules) • ________________– granules not attracted to stain • ________________– granules attracted to acidic stain • ________________– granules attracted to alkaline stain • ____________________(Absence of granules) • ________________ • ________________

  8. 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.

  9. 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.

  10. Details of the Granulocytes • Recall that the granulocytes include: • _____________________ • _____________________ • _____________________

  11. 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)

  12. 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.

  13. 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.)

  14. 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.

  15. 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)

  16. 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 ______________.

  17. 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 __________________.

  18. 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.

  19. 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 _____________.

  20. 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 _____________

  21. 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

  22. 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.

  23. 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 (__________________)

  24. Neutrophilia in a Periperal Blood Sample

  25. 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.

  26. 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

  27. 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 _________.

  28. 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. _________________, _______________)

  29. 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.

  30. 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

  31. 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

  32. Toxic Neutrophils Normal Neutrophils Toxic Neutrophils

  33. Toxic Neutrophils • Dohle Bodies may be noted in cytoplasm of toxic neutrophils. • Dohle Bodies are __________ and appear similar to _____________ seen in monocytes

  34. Left Shift

  35. 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.

  36. Hyper-segmented Neutrophil

  37. 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 ___________________________

  38. 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)

  39. Neutropenia in a Peripheral Blood Smear

  40. 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 ____________.

  41. Eosinophil Granules Feline Canine Equine Bovine

  42. 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.

  43. 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 ________.

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

  45. 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).

  46. Basophils Above: feline basophil (left); canine basophil (right)

  47. 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.

  48. 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.

  49. Agranulocytes • Recall that agranulocytes include • _________________________ • _________________________

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