Differential Leukocytes Flashcards
NUCLEUS composition of neutrophils
3 to 5 lobes separated by filaments
CYTOPLASM color of neutrophils
Pink to tan with violet or lilac granules
Most common WBC in normal peripheral blood
Neutrophils
First phagocytes to reach infection sites
Neutrophils, followed by monocytes
Neutrophil movement
Zigzag pattern, becomes straight line in presence of chemotactic attractant
Progenitor shared by neutrophils and monocytes
GMP (Granulocyte monocyte progenitor)
Major cytokine for neutrophil production
G-CSF (granulocyte colony stimulating factor)
Neutrophil half-life in blood
Approximately 7 hours
Major functions of neutrophils
1) Phagocytosis and destruction of foreign material, 2) Generation of NETs, 3) Secretory function (source of cytokines and transcobalamin I)
NETs structure
Extracellular threadlike structures made of chains of nucleosomes from unfolded nuclear chromatin (DNA)
NETs composition
Contain enzymes from neutrophil granules
NETs function
Trap and kill gram-positive and gram-negative bacteria, as well as fungi
NETs production timing
Produced when neutrophils die as a result of antibacterial activity
Unique form of neutrophil cell death that results in the release of NETs
NETosis
Order of formation of neutrophil granules
Primary, Secondary, Tertiary, Secretory
Order of degranulation of neutrophil granules
Secretory, Tertiary, Secondary, Primary
Size of neutrophil granules (largest to smallest)
Primary, Secondary, Tertiary, Secretory
Primary (Azyrophilic) Granules contents
Myeloperoxidase (MPO), Acid β-glycerophosphatase, Lysozyme, Cathepsins, Defensins, Elastase, Proteinase-3
Primary (Azyrophilic) Granules formation stage
Promyelocyte stage
Primary (Azyrophilic) Granules release order
Last to be released (exocytosis)
Myeloperoxidase function
Bactericidal, fungicidal, and viricidal properties
Myeloperoxidase detection method
Flow cytometry and cytochemical staining
Lysozyme function
Antimicrobial enzyme (muramidase)
Secondary (Specific) Granules contents
β2-microglobulin, Collagenase, Gelatinase, Lactoferrin, Neutrophil gelatinase-associated lipocalin, Transcobalamin I, Lysozyme
Secondary (Specific) Granules formation stage
Myelocyte and metamyelocyte stages
Secondary (Specific) Granules release order
Third to be released
Lactoferrin function
Iron-binding protein, competes with bacteria for iron
Tertiary (Gelatinase) Granules contents
Gelatinase, Collagenase, Lysozyme, Acetyltransferase, β2-microglobulin
Tertiary (Gelatinase) Granules formation stage
Metamyelocyte and band stages
Tertiary (Gelatinase) Granules release order
Second to be released
Secretory Granules contents
CD11b/CD18, Vesicle-associated membrane-2, CD10, CD13, CD14, CD16, Cytochrome b558, Complement 1q receptor, Alkaline phosphatase, Complement receptor-1
Secretory Granules formation stage
Band and segmented neutrophil stages
Secretory Granules release order
First to be released (fuse to plasma membrane)
Alkaline phosphatase activity in WBCs
Only neutrophils have alkaline phosphatase activity
CD11b/CD18 function in neutrophils
Contributes to tight stationary binding between neutrophils and endothelial cells
Neutrophil pools in the bone marrow
Stem cell pool, Mitotic (Proliferation) pool, Storage (Maturation) pool
Mitotic (Proliferation) pool components
CMP (CFU-GEMMs), GMP, Myeloblast, Promyelocyte, Myelocyte (last mitotic stage)
Storage (Maturation) pool components
Metamyelocyte, Band, Segmented neutrophils
Hematopoietic stem cell marker
CD34
Peripheral blood neutrophil pools and their ratio
Circulating Neutrophil Pool (CNP) and Marginal Neutrophil Pool (MNP), 50:50 ratio
Location of Circulating Neutrophil Pool (CNP)
Found in plasma
Location of Marginal Neutrophil Pool (MNP)
Found adhering to blood vessels
Nucleus characteristics of eosinophils
Dark purple, usually has two lobes
Cytoplasm characteristics of eosinophils
Filled with large, spherical granules of uniform size that stain bright orange
Most specific cytokine for eosinophil lineage
IL-5
Functions of IL-5 in eosinophil lineage
Promotes terminal maturation, functional activation, and prevention of apoptosis
Normal tissue destinations of eosinophils
Underlying columnar epithelial surfaces in respiratory, gastrointestinal, and genitourinary tracts
Hallmark condition associated with eosinophilia
Allergic disorders
Eosinophil’s role in helminthic infections
Secretes MBP, eosinophil cationic protein, and produces reactive oxygen species to destroy tissue-invading helminths
Eosinophil regulation of mast cells
Releases MBP to cause mast cell degranulation and cytokine production
Additional function of eosinophils in mast cell regulation
Produces nerve growth factor to promote mast cell survival and activation
Circulating half-life of eosinophils
Approximately 18 hours
Survival time of eosinophils in human tissues
2 to 5 days
Primary granules of eosinophils are formed during which stage?
Promyelocyte stage
Primary granules of eosinophils contain what key protein?
Charcot-Leyden crystal protein
Secondary granules of eosinophils are formed during which period?
Throughout remaining maturation
Core content of secondary eosinophil granules includes?
Major basic protein, Interleukins 2, 4, and 5, Granulocyte-macrophage colony-stimulating factor
Matrix content of secondary eosinophil granules includes?
Eosinophil cationic protein, Eosinophil-derived neurotoxin, Eosinophil peroxidase, Lysozyme, Catalase, Interleukin-6
Key functions of Charcot-Leyden crystal protein
Disintegration product of eosinophils
Appearance of Charcot-Leyden crystals under Hematoxylin stain
Black
Appearance of Charcot-Leyden crystals under Eosin stain
Red
Appearance of Charcot-Leyden crystals under Trichrome stain
Purplish red
Small lysosomal granules of eosinophils contain?
Acid phosphatase, Arylsulfatase B, Catalase, Cytochrome b558, Elastase, Eosinophil cationic protein
Lipid bodies in eosinophils contain enzymes like?
Cyclooxygenase, 5-Lipoxygenase, 15-Lipoxygenase, Leukotriene C4 synthase, Eosinophil peroxidase, Esterase
Function of storage (secretory) vesicles in eosinophils
Carry proteins from secondary granules to be released into the extracellular medium
Methods of eosinophil degranulation
1) Classical exocytosis, 2) Compound exocytosis, 3) Piecemeal degranulation, 4) Cytolysis
Granules fuse with the membrane and release contents
Classical exocytosis
Granules join together, then fuse with the membrane
Compound exocytosis
Vesicles take proteins from granules and release them
Piecemeal degranulation
Granules are released when the cell bursts
Cytolysis
Appearance of basophil nucleus
Difficult to observe due to overlying granules; generally unsegmented or bilobed
Appearance of basophil cytoplasm
Densely stained with dark violet to purple-black granules; variable in size and unevenly distributed
Contents of basophil granules
Histamine, Interleukin-4, Interleukin-13, Chondroitin sulfates (e.g., heparin)
Function of Toluidine blue stain
Binds acid mucopolysaccharides to form metachromatic complexes (e.g., reddish-violet granules in basophils and mast cells)
Metachromasia
Reaction product color different from the dye itself (e.g., reddish-violet granules with toluidine blue)
Fixative used before toluidine blue stain
Mota’s fixative
Value of toluidine blue stain
Identifies basophils and mast cells, especially neoplastic forms with reduced granules
Surface receptors on basophils
IgE receptors that trigger granule release upon cross-linking by antigen
Basophil-induced antibody synthesis
Induce B cells to synthesize IgE
Role of basophils in parasitic infections
Promote eosinophilia and aid worm expulsion
Least common WBC in peripheral blood
Basophils
Life span of basophils
60 hours
Mast cells vs basophils
Mast cells are connective tissue cells with basophilic granules (e.g., heparin, histamine) and are not leukocytes
Substances released by mast cells
Heparin, serotonin, bradykinin, histamine (in response to IgE stimulation)
Monocyte nucleus shape
Round, horse-shoe shaped or lobulated, with folding or convolutions
Monocyte chromatin appearance
Lace-like or stringy
Presence of nuclear vacuoles in monocytes
May be present
Monocyte cytoplasm appearance
Abundant, blue-gray with fine, indistinct granules (azure dust), ground-glass appearance
Features in monocyte cytoplasm
Small pseudopods or blebs, cytoplasmic vacuoles may be present
Monocyte size comparison
Largest cell in normal blood, about 2 to 3 times the diameter of an RBC
Monocyte identification stain
Strong positive reaction with nonspecific esterase stain
Enzymes used in nonspecific esterase stain
Alpha-naphthyl acetate and butyrate esterase
Sodium fluoride effect on monocytic enzyme
Inhibits enzyme activity of monocytic origin
Sodium fluoride effect on granulocytic or lymphocytic enzyme
No inhibition occurs
Monocyte enzyme present
Rich in muramidase
Promonocyte division under normal conditions
2 mitotic divisions to produce 4 monocytes in 60 hours
Promonocyte division under increased demand
4 divisions to produce 16 monocytes in 60 hours
Monocyte pools in peripheral blood
Marginal pool (3.5x the circulating pool) and circulating pool
Monocyte circulation time
Approximately 3 days
Characteristics of lymphocyte nucleus
Deep purple, round, oval, or indented with visible nucleoli
Typical color of lymphocyte cytoplasm
Sky-blue or ‘Robin egg’ blue
Sources of lymphocytes
Bone marrow, thymus, lymphatic system
Types of lymphocytes
T cells, B cells, Natural Killer (NK) cells
T & B cells immunity type
Adaptive immunity
NK cells immunity type
Innate immunity
Physiological characteristic 1 of lymphocytes
Predestined to migrate
Physiological characteristic 2 of lymphocytes
A heterogeneous group of cells
Physiological characteristic 3 of lymphocytes
Not an obligate end cell
Obligate end cell definition
A mature cell committed to perform a function, then dies (e.g., neutrophil)
Variant lymphocyte appearance
Nucleus: Dense to pale and immature-looking; Cytoplasm: Deeply basophilic to pale blue and abundant
Variant lymphocytes seen in
Nonmalignant reactive disorders
Synonyms for variant lymphocytes
Reactive lymphocytes, atypical lymphocytes, virocytes, stress lymphocytes, Downey cells, transformed lymphocytes, transitional lymphocytes, glandular fever cells
Type I variant lymphocyte
aka Turk’s Irritation Cell or Plasmacytoid Lymphocyte, seen in German Measles
Type II variant lymphocyte
aka Infectious Mononucleosis Cell, seen in Infectious Mononucleosis, characterized by flared skirt/fried egg appearance
Type III variant lymphocyte
Finely reticulated nuclear pattern