Leukocyte

Development, Kinetics
and Functions
GRANULOCYTES: Neutrophils
GRANULOCYTES: Neutrophils

*HSC, CMP and GMP are not distinguishable through staining and microscopy

Production rate: 0.9-1.0 x 109 cells/kg per day

Proliferation pool: contains approximately 2.1 x 109 cells/kg
Maturation pool: contains approximately 5.6 x 109 cells/kg (good
for 5 days supply)
GRANULOCYTES: Neutrophils
MYELOBLASTS
• About 0-3% of nucleated cells in the bone
marrow
• Measures 14-20um in diameter

1. Type I
- no visible granules
2. Type II
- with dispersed primary granules less
than 20 per cell
3. Type III
- rare in bone marrow and has darker
chromatin and cytoplasm with more than 20
granules per cell
GRANULOCYTES: Neutrophils
PROMYELOCYTES
• About 1-5% of the nucleated cells in the
bone marrow
• Measures 16-25 um in diameter
Nucleus: round to oval and eccentric
Cytoplasm: evenly basophilic and full of
primary (azurophilic) granules
GRANULOCYTES: Neutrophils
MYELOCYTES
• About 6-17% of the nucleated cells in
the bone marrow
• Measures 15-18 um in diameter
• Last stage capable of mitosis
• Stop of primary granules production
and start of secondary granules
GRANULOCYTES: Neutrophils
METAMYELOCYTES
• About 3-20% of the nucleated cells in
the bone marrow
• Measures 14-18 um in diameter
• Start of synthesis of tertiary granules
Nucleus: indented (kidney bean or
peanut shaped) with clumped chromatin
Cytoplasm: has very little residual RNA,
hence, less to no basophilia
GRANULOCYTES: Neutrophils
BANDS
• About 9-32% of the nucleated cells in
the bone marrow and 0-5% of the
nucleated peripheral blood cells
• Start of synthesis of secretory granules
Nucleus: highly clumped with
indentation more than half of the
nucleus without segmentation
Cytoplasm: absence of RNA, hence, no
cytoplasmic basophilia
GRANULOCYTES: Neutrophils
SEGMENTED NUETROPHILS
• About 7-30% of the nucleated cells
the bone marrow and 50-70% of th
nucleated peripheral blood leukocy
• Start of synthesis of secretory granu
Nucleus: presence of 2-5 nuclear lob
connected by thread-like filaments
Cytoplasm: same with bands

**Half-life: approximately 7 hours

 GRANULOCYTES: Neutrophils
PRIMARY GRANULES SECONDARY GRANULES TERTIARY GRANULES SECRETORY GRANULES
• Formed during • Formed during • Formed during • Formed during band and
promyelocyte myelocyte and metamyelocyte segmented stage
• Last to be released metamyelocyte and band stage • First to be released (fuse
stage • Second to be to plasma membrane)
• Third to be released released
• Myeloperoxidase • β2-macroglobulin • Gelatinase • CD 11b/ CD 18
• Acid β- • Collagenase • Collagenase • Alkaline phosphatase
glycerophosphatas • Gelatinase • Lysozyme • Vesicle-associated
e • Lactoferrin • Acetyltransferase membrane-2
• Cathepsins • Neutrophil • β2-macroglobulin • CD 10, CD 13, CD 14, CD
• Defensins gelatinase- 16
• Elastase associated lipocalin • Cytochrome b558
• Proteinase-3 • Trancobalamin I • Complement 1q
• Others • Others receptor
• Complement receptor-1
GRANULOCYTES: Neutrophils
GRANULOCYTES: Neutrophils
GRANULOCYTES: Eosinophils
• About 1-3% of the nucleated cells in the bone marrow and 1-3%
of the nucleated peripheral blood leukocytes

• GATA-1, PU.1, and c/EBP

GRANULOCYTES: Eosinophils
PROMYELOCYTE
- characterized by presence of
Charcot-Leyden crystals in primary
granules

MYELOCYTE
- characterized by presence of large,
pale, reddish-orange secondary granules,
along with azure granules in blue
cytoplasm
GRANULOCYTES: Eosinophils
METAMYELOCYTE and BANDS
- nuclear shape is same with that of
neutrophils
- secondary granules increase in
number
- start of secretory granules
synthesis
- characterized by 2 distinct
organelles:
a. lipid bodies
b. small granules
GRANULOCYTES: Eosinophils
MATURED EOSINOPHILS
Nucleus: bilobed
Cytoplasm: characterized by refractile,
orange-red secondary granules

**half-life:
In circulation – 18 hours
In tissues – 2-5 days

Turn-over rate: approximately 2.2 x 108 cells/kg per day

Storage pool (in the bone marrow): contains approximately 9-14
x 108 cells/kg
 GRANULOCYTES: Eosinophils
Type of Granules Formation Contents
Formed during Charcot-Leyden crystal protein
Primary
promyelocyte
Granules
stage
Formed Major Basic Protein, Eosinophil cationic
throughout protein, Eosinophil- derived neurotoxin,
Secondary
remaining Eosinophil peroxidase, Lysozyme, Catalase, β-
Granules
maturation glucuronidase, Cathepsin D, Interleukins 2, 4
and 5, GM-CSF
Acid phosphatase, arylsulfatase B, catalase,
Small Lysosomal
cytochrome b558, elastase, eosinophilic
Granules
cationic protein
Cyclooxygenase, 5-lipoxygenase, 15-
Lipid Bodies lipoxygenase, leukotriene C4 synthase,
esterase
GRANULOCYTES: Eosinophils
Functions:
1. Involved in the deletion of double positive thymocytes in
newborns
2. Acts as antigen presenting cells (APCs) and promotes proliferation
of effector T cells
3. Secretes cytokines that initiates of either type 1 or 2 immune
responses
4. Plays a role in acute and chronic allograft rejection
5. Regulate mast cell degranulation and survivability through MBP
release
6. Anti-helminthic response through MBP and eosinophilic cationic
proteins as well as reactive oxygen species
GRANULOCYTES: Basophils
• About <1% of the nucleated cells in the bone marrow and 0-2%
of the nucleated peripheral blood leukocytes
GRANULOCYTES: Basophils
IMMATURE BASOPHILS
Nucleus: have round to somewhat
lobulated nuclei with slightly condensed
chromatin
Cytoplasm: bluish cytoplasm and
contains blue-black secondary granules
GRANULOCYTES: Basophils
MATURE BASOPHILS
Nucleus: often obscured by its granules.
The chromatin pattern, if visible, is
clumped
Cytoplasm: usually colorless and contains
numerous large blue-black granules

*life span: 60 hours

GRANULOCYTES: Basophils
Functions:
1. Release Th2 immune response by secreting cytokines such as IL-4
and IL-13
2. Induces B cells to synthesize IgE
3. Initiates allergic inflammation through release of preformed
cytokines
4. Plays a role in angiogenesis through the expression of vascular
endothelial growth factor (VEGF) and its receptors
5. Controls helminthic infections, together with eosinophils, by
enclosing toxic egg products with granulomas and preventing
tissue damage
6. Play a nonredundant role in mediating acquired immunity against
ticks
MAST CELLS
 MAST CELLS
Functions:
1. Effector cells in allergic reactions
through IgE-dependent activation
2. Plays a role in inflammatory
processes
3. Plays as APCs to induce
differentiation of Th2 cells
4. Functions as anti-inflammatory and
immunosuppression
5. Act as immunologic “gatekeepers”
because of their location in mucosal
surfaces and role in barrier function
MONONUCLEAR CELLS: Monocytes
• About 2-11% of circulating leukocytes in the peripheral blood

Promonocyte pool: 6 x 108 cells/kg

Production rate: 7 x 106 cells/kg per hour
Life span: 3 days
 MONONUCLEAR CELLS: Monocytes
PROMONOCYTES
- measures 12-18 um in
diameter
Nucleus: slightly indented or folded
with delicate chromatin patterns
and at least one apparent nucleolus
Cytoplasm: bluish in color and
contains scattered azure granules
that are fewer and smaller than
those seen in promyelocytes
 MONONUCLEAR CELLS: Monocytes
MONOCYTES
- measures 15-20 um in diameter
Nucleus: may be round, oval, kidney
shaped, or deeply indented (horse-shoe
shaped) with loose chromatin patterns
described as lace-like or stringy
Cytoplasm: blue-gray in color with fine
azure granules referred as “azure dust” or
ground glass appearance. May be
characterized by cytoplasmic pseudopods
or blebs and vacuoles
 MONONUCLEAR CELLS: Monocytes

ORGAN MACROPHAGE
Liver Kupffer cells
Lungs Alveolar macrophages
Brain Microglia
Skin Langerhans cells
Spleen Splenic macrophages
Intestines Intestinal macrophages
Peritoneum Peritoneal macrophages
Bone Osteoclasts
Synovial macrophages Type A cells
Kidneys Renal macrophages
Reproductive organs Reproductive organ macrophages
Lymph nodes Dendritic cells
MONONUCLEAR CELLS: Monocytes
Functions:
1. Innate immunity
a. recognition of bacterial pathogens through toll-like receptors
b. synthesize nitric oxide, which is cytotoxic against viruses, bacteria,
protozoa, fungi, helminths and tumor cells
c. opsonin-dependent phagocytosis
2. Adaptive immunity
- functions as antigen-presenting cells
3. Housekeeping functions
a. removal of debris and dead cells at sites of infection or inflammation
b. destruction of senescent red cells and maintenance of storage pool
of iron for erythropoiesis
c. synthesis of wide array of proteins like coagulation factors, IL and
enzymes
MONONUCLEAR CELLS: Lymphocytes
• Three major groups: T cells, B cells, and NK cells
• About 18-42% of circulating lymphocytes

Antigen-independent development:
- happens in central or primary
lymphatic organs
Ex. Bone marrow and Thymus

Antigen-dependent development:
- happens in peripheral or
secondary lymphatic organs
Ex. Spleen, lymph nodes, tonsils, MALT
 MONONUCLEAR CELLS: Lymphocytes

Pro-B Pre-B Immature

cells cells B cells

Functions:
1. Antibody production
2. Antigen presentation to T cells
3. Cytokine production for regulation
of T cells and APC functions
 MONONUCLEAR CELLS: Lymphocytes
Functions:
T helper (CD4+) cells
Th1 – response against intracellular pathogens
Th2 – defense against extracellular parasites
Th17 – response against extracellular bacteria and fungi
Treg – maintains self tolerance by regulating immune
response
T cytotoxic (CD8+) cells
Response to intracellular pathogens and tumors cells

Pro-T Pre-T Immature

cells cells T cells
 MONONUCLEAR CELLS: Lymphocytes
NATURAL KILLER (NK) CELLS
- approximately about 4-29% of
circulating lymphocytes
- surface antigens: CD 56, CD16, and
CD7

Functions:
1. Kills virally infected cells and tumor
surveillance
2. Modulate functions of other cells
including macrophages and T cells
WBC COUNTING and DIFFERENTIALS
DIFFERENTIAL COUNTING METHODS
1. Cross-sectional or crenellation method
2. Longitudinal method CELL ABSOLUTE COUNT RELATIVE COUNT
3. Battlement method
Neutrophils 2.3-8.1 x 109/L 50-70%
Bands 0-0.06 x 109/L 0-5%
Eosinophils 0-0.4 x 109/L 1-3%
Basophils 0-0.2 x 109/L 0-2%
Monocytes 0.01-1.3 x 109/L 2-11%
Lymphocytes 0.8-4.8 x 109/L 18-42%
 CELL ABSOLUTE COUNT RELATIVE COUNT
Neutrophils 2.3-8.1 x 109/L 50-70%
Bands 0-0.06 x 109/L 0-5%
Eosinophils 0-0.4 x 109/L 1-3%
Basophils 0-0.2 x 109/L 0-2%
Monocytes 0.01-1.3 x 109/L 2-11%
Lymphocytes 0.8-4.8 x 109/L 18-42%
WBC counting and differentials

NEUTROPHILIA NEUTROPENIA
• Physiologic stress (exercise, acute • Viral infections
emotional stress, exposure to heat and • Exposure to radiation, cytotoxic drugs
cold) and benzene
• Systemic or severe local infections • Liver disease
• Acute rheumatic fever • Vitamin B12 deficiency
• Gout
• Exposure to epinephrine, lithium,
histamine , heparin
WBC counting and differentials
EOSINOPHILIA
• Allergic reactions
• Parasitism
• Neoplastic diseases (Hodgkin’s
lymphoma)
• hypoadrenalism
BASOPHILIA
• Systemic mast cell disease
• Chronic hypersensitivity
• Hypothyroidism
• Ulcerative colitis
• Estrogen therapy
• Myeloproliferative disorders
EOSINOPENIA
• Cushing’s syndrome
• Labor and eclampsia
• After electric shock treatment
BASOPENIA
• Acute infections
• Stress
• Hyperthyroidism
• Increased levels of glucocorticoids
WBC counting and differentials
LYMPHOCYTOSIS LYMPHOPENIA
• Whooping cough, brucellosis, syphilis • Immunodeficiency syndromes
• Viral infections • Adrenal gland hyperactivity
• Hypoadrenalism
• Immune diseases

MONOCYTOSIS MONOCYTOPENIA
• Tuberculosis • Increased levels of glucocorticoids
• Chronic bacterial infections
• Cancers, lymphomas and
myeloproliferative disorders