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Hematology and Cytology > Leukon #1: Morphology and Function > Flashcards

Leukon #1: Morphology and Function Flashcards

1
Q

Neutrophil - Size

A

~11 microns

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2
Q

Neutrophil - Nucleus

A
  • Segmented 2-4 lobes
  • Dark, purplish to almost black
  • Condensed, “mature” chromatin
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3
Q

Neutrophil - Cytoplasm

A
  • Clear to faint pink

- Feline - slight blue

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4
Q

Neutrophil - Inclusions

A
  • Granules do not stain (neutral) or light pink
  • Two main types of granules (lysosomes)
  • -Primary (or azurophilic)
  • -Secondary (or specific)
  • -(Also Tertiary - less important)
  • Granule contents:
  • -Sequestered in granules to protect the cell
  • -Secreted into phagocytic vacuoles for killing
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5
Q

Neutrophil - Barr Body

A
  • Sex chromatin lobe
  • Low percentage of neutrophils of female mammals
  • Inactivated X chromosomes
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6
Q

Primary Granules

A
  • Small reddish-purple granules
  • Similar to lysosomes
  • Contain acid hydrolases
  • Contain myeloperoxidase (MPO)
  • -Neutrophils can be marked based on presence of MPO
  • Give neutrophils their function
  • BM: formed at late myeloblast/ early promyelocyte stage
  • Don’t normally stain very well in blood smears
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7
Q

Secondary Granules

A
  • Stain neutral
  • Specific granules
  • Most numerous
  • Contents involved in the inflammatory response
  • ID the cell
  • BM: develop at myelocyte stage
  • -Other granulocytes uptake stain based on pH
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8
Q

Tertiary Granules

A
  • Contain secretory enzymes

- Can insert adhesion molecules on the surface of the cell

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9
Q

Granule Considerations

A
  • Fine/faint pink granulation common
  • Mostly artifact: pH, sample age
  • Healthy neonatal foals: healthy
  • Toxic granulation
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10
Q

Granulopoiesis - Maturation

A
  • Myeloblasts have nucleoli
  • As they mature
  • -Size - smaller
  • -Cytoplasm - less basophilic
  • -Nucleus - more lobulated
  • -Chromatin - more coarse and condensed
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11
Q

Neutrophil - Function

A
  • Primary function: Phagocytosis and Bacterial Killing
  • -Respond to strong irritants
  • –Bacterial products, chemical toxins, tissue injury
  • Metabolically active processes
  • -Adhere and transmigrate through vessel wall
  • -Chemotaxis (migration along chemical gradient)
  • -Phagocytosis (ingestion) - forms phagosome
  • -Degranulation - into phagosome or externally
  • -Killing
    1. Granule contents (microbiocidal, enzymes)
    2. Respiratory burst - toxic reactive oxygen species
  • Also assist with lower grade irritants:
  • -Fungal organisms, yeasts, algae, parasites, viruses, foreign bodies
  • Receptors
  • -Cytokines
  • -Toll-like receptors
  • -Pathogen products (LPS, f-MLP)
  • -Adhesion molecules (selectins, integrins)
  • Activation of NADPH oxidase and microbial systems during phagocytosis
    1. Complement and antibody receptors promote
    2. Uptake of microorganisms
    3. Degranulation
    4. Production of ROS
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12
Q

Eosinophil - Size

A

~12-14 microns

  • Greater than size of neutrophil
  • Less than or equal to size of basophil
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13
Q

Eosinophil - Nucleus

A
  • 2-3 round to irregular lobes

- Chromatin dark, condensed

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14
Q

Eosinophil - Cytoplasm

A

-Clear, faint pink/blue

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15
Q

Eosinophil - Inclusions

A
  • Secondary granules differ by species
  • -Dog: round, variable sizes
  • -Sighthounds: gray granules
  • -Cats: rod-shaped granules
  • -Horse: large and round
  • -Ruminants and pigs: small and round
  • Secondary granule contents:
  • -Major basic protein - cytotoxic to parasites, protozoa, and bacteria
  • -Eosinophil peroxidase - generates ROS toxic to helminths, bacteria, mycoplasmas, fungi, protozoa, viruses, and tumor cells
  • -Eosinophil cationic protein - toxic to helminths, protozoa, and bacteria
  • -Eosinophil-derived neurotoxin - toxic to myelinated nerve fibers
  • Tertiary granules:
  • -Contain secretory enzymes
  • -Can insert adhesion molecules on the surface of the cell
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16
Q

Eosinophil - Function

A
  • Association: worms, wheezes, and weird diseases
  • -IgE mediated conditions (released by plasma cells)
  • -Recruited to the tissues by IL-5 (released from Th2 cells)
  • Helminths
  • -Eosinophils bind to and degranulate
  • -Release major basic protein
  • -Release eosinophil-specific peroxidase
  • -Production of ROS
  • -Aided by complement and T-cell perforins
  • Hypersensitivity
  • -Attracted by mast cell/basophil chemicals
  • –Ex: histamine (respond with release of anti-histamine)
  • -Can promote inflammation, tissue damage
  • –Allergies, asthma - bind IgE-Ag and exocytose
  • –Inflammation, fibrosis, etc.
  • -Phagocytosis (possible, but not as efficient at it as neutrophils are)
  • Tumor associated eosinophilia
  • -Often paraneoplastic by IL-5 production by tumor cells
  • -Ex: mast cell tumor, T-cell lymphoma, carcinoma, fibrosarcoma
  • Rapid blood transit time
  • Long life in the tissues: lungs, GI tract, and skin
17
Q

Basophil - Size

A

~14-15 microns

  • Greater than or equal to size of eosinophils
  • Less than or equal to size of monocyte
18
Q

Basophil - Nucleus

A
  • 2-3 lobes, may look ribbon like
  • Medium to dark chromatin
  • Varies from seg to monocyte-like chromatin density
19
Q

Basophil - Cytoplasm

A

-Clear, blue/gray, magenta blue

20
Q

Basophil - Inclusions

A
  • Secondary granules vary by species
  • -Dog: indistinct lavender to purple granules
  • -Cat: pale to distinct lavender, rod-shaped
  • -Horse and ruminant: purple
21
Q

Basophile - Functions

A
  • Associated with similar conditions as eosinophils (worms, wheezes, and weird diseases)
  • Associated with similar functions as mast cell (similar granule contents)
  • -Histamine: both
  • -Heparin (involved in hemostasis): mast cells
  • -Share a progenitor in marrow, but…
  • –Different nuclear morphology
  • –Basophils leave marrow mature
  • –Mast cells leave marrow agranular
  • –Mast cells mature in tissues (rarely seen in blood)
22
Q

Mononuclear cell - General info

A
  • NOT devoid of granules
  • -Granules still present, just fewer - less obvious
  • -Large Granular Lymphocytes (LGLs)
  • –Focal pink/purple granules
  • –May be cytotoxic T cells or NK cells
23
Q

Monocytes –> Macrophages

A

Monocytes become macrophages and dendritic cells in the tissues

  • Collectively called Mononuclear Phagocyte System (MPS)
  • -Includes:
  • –Blood monocyte-derived macrophages
  • –Dendritic cells and fixed
  • –Tissue-specific cells
24
Q

Monocyte - Size

A

Greater than or equal to 15 microns

25
Q

Monocyte - Nucleus

A
  • Oval, reniform, bilobed, trilobed

- Light, lacy chromatin, some clumped

26
Q

Monocyte - Cytoplasm

A

-Light to dark blue-gray

27
Q

Monocyte - Inclusions

A
  • Vacuoles common
  • -Round, variable sizes, ‘punched out’ v. foamy
  • -Some may be sample age/storage induced
  • Fine pink granules possible (lysosomes)
28
Q

Monocyte - Tissue Differentiation

A
  • Blood monocyte-derived macrophage activation leads to specialization:
  • -Phagocytic cells (classic macrophage)
  • –Increased vacuolated cytoplasm
  • –Enhanced phagocytosis, clean up
  • –Organisms, debris, foreign material
  • -Epithelioid cells (signaling)
  • –Orchestrate inflammation through secretory functions
  • Blood monocyte-derived dendritic cells
  • -APCs in various tissue sites
  • –Skin (Langerhans cells)
  • –Mucosal sites: respiratory, digestive, etc.
  • –Lymphoid tissues
  • –Link innate and adaptive immune systems
  • Fixed tissue macrophages
  • -Ex: Kupffer cells (liver), microglia (CNS), alveolar cells (lungs), osteoclasts (bone), etc.
  • -Long lived (weeks/months)
  • -Thought all were blood monocyte derived
  • -Recent reports suggest these may be seeded in sites early in life and replicate there
29
Q

Monocyte/Macrophage - Functions

A

-Respond to low-grade irritants (v. neutrophil)

  • Phagocytize: foreign materials, dead/effete cells
  • -Clean up foreign and domestic debris
  • -Not as efficient as neutrophils on bacteria
  • –Higher order bacteria: mycobacteria, nocardia, actinomyces
  • –Protozoa: leishmania, toxoplasma
  • –Fungi: balstomycosis
  • -Kill cell function (immune)
  • –Ab-mediated cytotoxic function (viral-mediated, tumor cells)
  • Secrete: orchestrate inflammation and tissue repair
  • -Inflammation
  • –Chemotaxis factors, complement, collagenase, elastase, etc.
  • –G-CSF, GM-CSF:
  • —PMNs: increase marrow divisions, inhibit apoptosis, “prime” functions
  • -Hematopoiesis
  • –Major source of CSFs, cytokines
  • —G-CSF, M-CSF, IL-1, IL-3, TNF
  • –Iron metabolism –> erythropoiesis
30
Q

Lymphocyte - Size

A

~8-10 microns (normal and small)

  • High NC ratio
  • Bigger than RBC, smaller than neutrophil
  • Ruminants: small and large (less than or equal to 15 microns)
  • Sizing lymphocytes:
  • -Small: nucleus fits inside neutrophil
  • -Intermediate: nucleus is same size as neutrophil
  • -Large: neutrophil fits inside nucleus
31
Q

Lymphocyte - Cytoplasm

A
  • Clear to faint blue

- Darker blue edges if reactive

32
Q

Lymphocyte - Nucleus

A
  • Round to slightly angular
  • Ruminant: large lymphocytes more irregular
  • Stimulated neoplastic lymphocytes –> +/- pleomorphic
  • Moderately condensed chromatin
33
Q

Lymphocyte - General Information

A
  • Morphology alone only IDs plasma cells and LGLs
  • Cell markers (CD markers) are needed to subtype lymphocytes
  • Have a heterogeneous population
  • Lymphocytes recirculate
34
Q

Lymphocyte - Inclusions

A
  • Granular lymphocytes (should only be ~10% granular)
  • -Small pink or purple granules
  • -Often in perinuclear clear area
  • -Mostly cytotoxic T or NK cells
  • -Can be normal, reactive, or neoplastic
35
Q

Lymphocyte - Artifacts

A

Sitting in EDTA 30-60 minutes

  • Possible vacuoles, lobulation, or smudging of nuclei
  • Make smears ASAP after drawing blood
  • True in other cells to varying degrees
36
Q

Lymphocyte - Functions

A
  • B cells
  • -Humoral (antibody) immunity
  • -Divide and transform into effector cells that produce IG
  • -Plasma cells are the most differentiated cell
  • T cells
  • -Cell-mediated immunity
  • -Divide and transform into effector cells that produce lymphokines and mediate cellular immunity

Hematology and Cytology (6 decks)

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