GRANULOCYTIC

SERIES
 Cellular elements of the blood
- produce from a common, multipotential hematopoietic cell

Progenitor Cell
- undergoes mitotic division

The major categories of the cellular elements of the

circulating blood:
Erythrocytes
Leukocytes
Thrombocytes

Basis of function
LEUKOCYTES
granulocytic
monocytic
lymphoid series
Granulocytes
are a group of leukocytes whose cytoplasm is filled with granules with
differing staining characteristics and whose nuclei are segmented or
lobulated.
Eosinophils
granules containing basic proteins that stain with acid stains such as eosin
Basophils
granules that are acidic and stain with basic stains such as methylene blue
Neutrophils
granules that react with both acid and basic stains, which gives them a
pink to lavender color.
 GARNULOCYTIC SERIES
Production of Neutrophils, Eosinophils, and Basophils

Cells that are committed to differentiation (granulocytes)

- cloned in vitro
- produced a mixture of granulocytes and macrophage cells
Dependent on
colony-stimulating factor (CSF)
interleukins

- presence of different CSFs favors interleukins

- the microenvironment of the progenitor cell favors differential development of
either the granulocytic (myeloid) series or the macrophage-monocytic series.
- different CFSs stimulate specific differentiation - development of eosinophils
 GARNULOCYTIC SERIES
Sites of Development and Maturation

GRANULOCYTIC KINETICS

- development, distribution, and destruction of neutrophils, eosinophils,

and basophils.
MULTIPOTENTIAL CELL (in bone marrow)
Neutrophil
Basophil
Eosinophil

- remain in the bone marrow

- after developing (band or segmented forms)
- mature cells enter into the blood circulation
 GARNULOCYTIC SERIES

Neutrophils
-Neutrophils are present in the peripheral blood in two forms according to whether the nucleus
is segmented or still in a band shape. Segmented neutrophils make up the vast majority of
circulating leukocytes.

Neutrophil Development
-Neutrophil development occurs in the bone marrow. Neutrophils share a common progenitor
with monocytes and distinct from eosinophils and basophils, known as the
granulocytemonocyte progenitor (GMP).
-The major cytokine responsible for the stimulation of neutrophil production is granulocyte
colony-stimulating factor, or G-CSF.
 GARNULOCYTIC SERIES

Eosinophils
-Eosinophils make up 1% to 3% of nucleated cells in the bone marrow.
.-Slightly more than a third are mature, a quarter are eosinophilic metamyelocytes, and the

remainder are eosinophilic promyelocytes or eosinophilic myelocytes.

Eosinophil Development
-Eosinophil development is similar to that described earlier for neutrophils, and evidence
indicates that eosinophils arise from the CMP.
 GARNULOCYTIC SERIES

Basophils
-Basophils and mast cells are two cells with morphologic and functional similarities
-basophils are true leukocytes because they mature in the bone marrow and circulate in the
blood as mature cells with granules
-mast cell precursors leave the bone marrow and use the blood as a transit system to gain
access to the tissues where they mature.

Basophil Development
-Basophils are derived from progenitors in the bone marrow and spleen, where they
differentiate under the influence of a number of cytokines, including IL-3 and TSLP (thymic
stromal lymphopoietin)
 GARNULOCYTIC SERIES
There are three pools of developing neutrophils in the bone marrow
Stem cell pool
-consists of HSCs that are capable of self-renewal and differentiation.

.Proliferation (mitotic) pool

-consists of cells that are dividing and includes (listed in the order of maturation) common
myeloid progenitors (CMPs), also known as colony-forming units– granulocyte, erythrocyte,
monocyte, and megakaryocyte (CFUGEMMs); granulocyte-monocyte progenitors (GMPs);
myeloblasts; promyelocytes; and myelocytes.

Maturation (storage) pool

-consisting of cells undergoing nuclear maturation that form the marrow reserve and are
available for release: metamyelocytes, band neutrophils, and segmented neutrophils.
 GARNULOCYTIC SERIES
Development and Proliferation of Neutrophils, Eosinophils, and Basophils

colony-forming-unit-granulocyte-erythrocyte-megakaryocyte (CFU-GEMM) progenitor

cell differentiates into the colony-forming-unit-granulocyte-macrophage (CFU-GM)
progenitor cell

- the cell line becomes committed to developing into a myeloblast

- the maturational development from the myeloblast through the myelocyte
stage and mitotic division take place - bone marrows proliferative
compartment.
- mitotic pool and includes cells capable of DNA synthesis
 GARNULOCYTIC SERIES
Development and Proliferation of Neutrophils, Eosinophils, and Basophils

MYELOBLAST
- the first identifiable cell in the granulocytic series
- constitute approximately of the total nucleated bone
marrow cells
- lasts approximately 15 hours

PROMYELOCYTE
- constitutes approximately 3% of the nucleated bone
marrow cells
- lasts about 24 hours
 GARNULOCYTIC SERIES
Development and Proliferation of Neutrophils, Eosinophils, and Basophils

MYELOCYTE
- with approximately 12% of the proliferative cells
existing in this stage

METAMYELOCYTE
- lasts an average of 4.3 days
- metamyelocyte stage has been reached,

cells have undergone four or live cell divisions

Proliferative Phase comes to an end

 GARNULOCYTIC SERIES
Development and Proliferation of Neutrophils, Eosinophils, and Basophils

GRANULOCYTES - enter a maturation-storage

compartment

METAMYELOCYTES AND BAND FORMS

- mature into segmented granulocytes in this
compartment of the bone marrow.

MARROW RESERVE
- the segmented neutrophils in the maturation-
storage compartment
- constitutes a 4- to 8-day supply of neutrophils
 GARNULOCYTIC SERIES
Development and Proliferation of Neutrophils, Eosinophils, and Basophils

NEUTROPHILIC GRANULOCYTES
- normally remain in the maturation-storage phase for 7 to 10 days

EOSINOPHILS
- remain for about 2.5 days

BASOPHILS
- remain in this phase for the shortest period approximately 12 hours
 GARNULOCYTIC SERIES
Distribution of Neutrophils, Eosinophils, and Basophils

release of neutrophils from the bone marrow into the

circulatory system = COMPLEX PROCESS

Characteristics and Physiological Regulators

- promote movement of the granulocytes (sinusoid
wall of the bone marrow) anatomical barrier
 GARNULOCYTIC SERIES
Distribution of Neutrophils, Eosinophils, and Basophils

Cellar Characteristics
- overall reduction in cell size
- a smaller nuclear-cytoplasmic ratio

greater flexibility and mobility of mature cells

- enhance the migration of cells (marrow sinusoids)
- peripheral blood pool

Peripheral Blood Circulation pools of equal size:

Circulating Pools
Marginating Pools
 GARNULOCYTIC SERIES
Distribution of Neutrophils, Eosinophils, and Basophils
Marginating Granulocytes
- adhere to the endothelium of the blood vessels.
- found in the spleen

Mature granulocytes (peripheral blood)

- only in transit to their potential sites of action in
the tissues

DIAPEDESIS
- movement of granulocytes from the circulating
pool to the peripheral tissues

Peripheral Tissues (NEUTROPHILS)

- able to carry out their function of phagocytosis
 GARNULOCYTIC SERIES
Distribution of Neutrophils, Eosinophils, and Basophils
SEGMENTED NEUTROPHILIC GRANULOCYTE (circulating blood)
average lifespan:
- approximately 7 to 10 hours

Mature Cells (migrated into the tissues)

- life span is considered to be several days
unless the cells encounter antigens, toxins, or
microorganisms

Eosinophils (peripheral blood)

- for a few hours believed to reside in the tissues for
several days
 GARNULOCYTIC SERIES
Distribution of Neutrophils, Eosinophils, and Basophils

Basophils
average circulation time of about 8.2 hours.

excessive numbers of eosinophils

disease state
damaged or degenerated eosinophils
Charcot-Leyden crystals (body secretions)
sputum
stool
 GARNULOCYTIC SERIES
Distribution of Neutrophils, Eosinophils, and Basophils

CELLS (not prematurely destroyed while defending the body)

sloughed off with various body secretions
Urine
Saliva
Gastrointestinal secretions

PHAGOCYTOSIS
alternative route for the removal of granulocytes from the circulation
by the mononuclear phagocyte cells of the spleen
DE GUZMAN
BMLS 3-A
 INTRODUCTION
The cellular elements of the blood are produced from a common, multipotential
hematopoietic cell. This cell, the progenitor cell, undergoes mitotic division.
Subsequent maturation of progenitor cells produces the major categories of the
cellular elements of the circulating blood: the erythrocytes, leukocytes, and
thrombocytes.
• GRANULOCYTES

• EOSINOPHILS
Granules containing basic proteins that stain with acid stains such as eosin;

• BASOPHILS
Granules that are acidic and stain with basic stains such as methylene blue;

• NEUTROPHILS
Granules that react with both acid and basic stains, which gives them a pink to lavender
color. Because nuclear segmentation is quite prominent in mature neutrophils, they have
also been called polymorphonuclear cells, or PMNs.
 GRANULOCYTIC SERIES: PRODUCTION OF
NEUTROPHILS, EOSINOPHILS, AND BASOPHILS

The development of blood cells from hematopoietic stem cells in the bone marrow is a complex
process that depends on the microenvironment consisting of stromal cells, cytokines, and the
extracellular matrix.

Cytokines mediate communication between the cells and exert their biological functions mostly
through specific receptors expressed on the surface of the target cells. These signal substances have
the capacity to stimulate, enhance, or suppress the proliferation of the stem and progenitor cells.

Cells that are committed to differentiation as granulocytes have been cloned in vitro and have
produced a mixture of granulocytes and macrophage cells.
 GRANULOCYTE SITES OF
GROWTH FACTORS DEVELOPMENT AND
MATURATION
• These cells is dependent on colony-
The development, distribution, and
stimulating factor (CSF) and
destruction of neutrophils, eosinophils,
interleukins.
and basophils are collectively referred to
• The presence of different CSFs favors as granulocytic kinetics or the cell
interleukins, and the microenvironment turnover rate. The neutrophil, basophil,
of the progenitor cell favors differential and eosinophil each begin as a
development of either the granulocytic multipotential cell in the bone marrow.
(myeloid) series or the macrophage- Throughout the normal processes of
monocytic series. differentiation, multiplication, and
maturation, these cells remain in the bone
• In addition to the differentiation of
granulocytes and monocytes, different marrow. After developing into their band
CSFs stimulate specific differentiation, and segmented forms, mature cells enter
such as the development of into the blood circulation
eosinophils.
 NEUTROPHIL
• Neutrophils are present in
the peripheral blood in two
forms according to whether
the nucleus is segmented
or still in a band shape.
Segmented neutrophils
make up the vast majority
of circulating leukocytes
THREE POOLS OF
DEVELOPING NEUTROPHILS
IN THE BONE MARROW
NEUTROPHIL
DEVELOPMENT
• Occurs in the bone marrow
• Share a common progenitor with
monocytes and distinct from
eosinophils and basophils, known as
the granulocyte monocyte progenitor
(GMP)
• The major cytokine responsible for
the stimulation of neutrophil
production is granulocyte colony-
stimulating factor, or G-CSF
• STEM CELL POOL
• PROLIFERATION (MITOTIC) POOL
• MATURATION (STORAGE) POOL
THREE POOLS OF DEVELOPING NEUTROPHILS IN THE BONE MARROW

• STEM CELL
POOL
- Consists of HSCs that are capable of self-renewal and differentiation.

• PROLIFERATION (MITOTIC) POOL

- Consists of cells that are dividing and includes (listed in the order of maturation) common
myeloid progenitors (CMPs), also known as colony-forming units– granulocyte, erythrocyte,
monocyte, and megakaryocyte (CFUGEMMs); granulocyte-monocyte progenitors (GMPs);
myeloblasts; promyelocytes; and myelocytes.

• MATURATION (STORAGE) POOL

- Consisting of cells undergoing nuclear maturation that form the marrow reserve and are
available for release: metamyelocytes, band neutrophils, and segmented neutrophils.
NEUTROPHIL DEVELOPMENT SHOWING STIMULATING CYTOKINES AND THE THREE
BONE MARROW POOLS
NEUTROPHIL DEVELOPMENT SHOWING STIMULATING CYTOKINES AND THE
THREE BONE MARROW POOLS

• HSCs, CMPs, and GMPs are not distinguishable with the light microscope and Romanowsky staining and
may resemble early type I myeloblasts or lymphoid cells. They can, however, be identified through surface
antigen detection by flow cytometry.

• The MYELOBLAST is the first identifiable cell in the granulocytic series. Myeloblasts constitute approximately
1% of the total nucleated bone marrow cells. This stage lasts approximately 15 hours.

• The next stage, the promyelocyte, constitutes approximately 3% of the nucleated bone marrow cells. This
stage lasts about 24 hours.

• The myelocyte is the next maturational stage, with approximately 12% of the proliferative cells existing in this stage.

• The stage from myelocyte to metamyelocyte lasts an average of 4.3 days. Once the metamyelocyte stage has
been reached, cells have undergone four or five cell divisions and the proliferative phase comes to an end.
 MATURATION-STORAGE COMPARTMENT
• The metamyelocytes and band forms
mature into segmented granulocytes in the compartment of the bone
marrow.

• The relative proportions of these cells are approximately 45%,

35%, and 20%, respectively. The segmented neutrophils in the
maturation-storage compartment are frequently referred to as the marrow
reserve.

• This reserve constitutes a 4- to 8-day supply of neutrophils. It is

estimated that neutrophilic granulocytes normally remain in the
maturation-storage phase for 7 to 10 days.

• Eosinophils remain for about 2.5 days, and basophils remain in this phase for the shortest period, approximately
12 hours.
• From this stage forward, the cells are no longer capable of division and the major morphologic change is in
the shape of the nucleus. The nucleus is indented (kidney bean shaped or peanut shaped), and the chromatin
is increasingly clumped. Nucleoli are absent. Synthesis of tertiary granules (also known as gelatinase
granules) may begin during this stage.
 DISTRIBUTION OF NEUTROPHILS,
EOSINOPHILS, AND BASOPHILS

• The release of neutrophils from the bone marrow into the

circulatory system is a complex process.

• Certain characteristics and physiological regulators promote

movement of the granulocytes through the sinusoid wall of the
bone marrow, which is normally an anatomical barrier.

• Some of the factors that influence cellular release include the

interleukins. Cellular characteristics include an overall reduction in
cell size and a smaller nuclear-cytoplasmic ratio.

• The greater flexibility and mobility of mature cells enhance the

migration of cells through the marrow sinusoids into the peripheral
blood pool.
COMPARATIVE MATURATIONAL
CHARACTERISTICS.
The peripheral blood circulation is subsequently divided into two pools
of equal size:
• CIRCULATING POOL
• MARGINATING
POOL.
The marginating granulocytes adhere to the endothelium of the blood vessels Some granulocytes are
additionally found in the spleen. Mature granulocytes in the peripheral blood are only in transit to their
potential sites of action in the tissues.

DIAPEDESIS - • The movement of granulocytes from the circulating pool to the peripheral tissues
• Once in the peripheral tissues, granulocytes, particularly the neutrophils, are able to
carry out their function of PHAGOCYTOSIS.

LIFE SPAN IN CIRCULATING POOL:

• The average life span of a segmented neutrophilic granulocyte in the
circulating blood is approximately 7 to 10 hours.
• Eosinophils are in the peripheral blood for a few hours and are believed to reside in the tissues for
several days.
• Basophils have an average circulation time of about 8.5 hours.
CHARCOT-LEYDEN
crystals
• If excessive numbers of eosinophils are present because of a disease
state, damaged or degenerated eosinophils give rise to found in body
secretions, such as the sputum and stool.
• If cells are not prematurely destroyed while defending the body, they are
sloughed off with various body secretions, such as the urine, saliva, or
gastrointestinal secretions. An alternative route for the removal of granulocytes
from the circulation is phagocytosis by the mononuclear phagocyte cells of the
spleen
NORMAL MATURATIONAL CHARACTERISTICS OF
NEUTROPHILS, EOSINOPHILS, AND BASOPHILS
 • TYPE I (LEFT CELL)
• TYPE II (RIGHT CELL)
MYELOBLASTS

MYELOBLAST
• The earliest morphologically identifiable granulocytic
precursor is the myeloblast

• This cell has an average overall diameter of 10 to 18 μm. The

nuclear chromatin is finely reticular, with one to five light-
staining nucleoli. The cytoplasm appears as a small rim of
basophilic cytoplasm that lacks granules.
NEUTROPHILIC SERIES
 AUER RODS
Auer rods are aggregates of fused lysosomes, may
appear as red, needle-like crystalline cytoplasmic
inclusions. These inclusions may appear alone or in
groups. Auer rods are pathological, not normal,
inclusions.

AUER RODS (ARROW)

 PROMYELOCYTE
• The promyelocyte represents the second maturational stage seen in
granulocytes. The outstanding feature of this cell is the
presence of prominent granulation that may actually obscure the
other morphological features of the cell. These granules are primarily
azurophilic granules and are rich in the enzymes myeloperoxidase
(MPO) and chloroacetate esterase.

• The promyelocyte is larger than theblast stage, with an average

diameter of 14 to 20 μm. The N:C ratio is lower in the promyelocyte
than in the myeloblast. The nuclear chromatin is more condensed
than in the blast, and nucleoli are present. The
cytoplasm is a pale grayish blue.

PROMYELOCYTES. (A), PROMYELOCYTE (ARROW) WITH

NUCLEOLI
AND A LARGE NUMBER OF AZURE GRANULES.
(B), ELECTRON MICROGRAPH OF A PROMYELOCYTE
 MYELOCYTE

• The myelocyte is the third maturational stage. This cell is

• The myelocyte has an average diameter of 12 to 18 μm. The N:C
. (A), TWO EARLY NEUTROPHIL MYELOCYTES. (B), ARROWS ARE POINTING TO THREE LATE
MYELOCYTES IN THE FIELD.
characterized by the recognizable appearance of secondary or
specific cytoplasmic granulation. The separate cell types—
neutrophils, eosinophils, and basophils—become visibly
recognizable at this stage. Neutrophilic granules are fine and stain
a blue-pink color with Wright stain. Eosinophilic granules are
larger than neutrophilic granules. These round or oval-shaped
granules are orange and have a glassy or semiopaque texture.
Basophilic granules have a dark blue-black color and a dense
appearance.
ratio continues to decrease. The nucleus has a more oval
appearance than in previous stages, nucleoli are no longer visible,
and the chromatin is much more clumped than in previous stages.
 METAMYELOCYTE

• The metamyelocyte is the fourth maturational stage. Its

most characteristic feature is that the nucleus begins to
assume an indented or kidney bean shape, which will
continue to elongate as the cell matures through this
phase. The chromatin continues to become more
condensed or clumped.

• The color of the specific granulation continues to become

a major distinguishing feature. In human bone marrow,
METAMYELOCYTES. (A), TWO NEUTROPHIL METAMYELOCYTES
the metamyelocyte is the most common granulocytic
(ARROWS).
leukocyte.
 MATURE FORMS

• Two stages of granulocytes are observed in the

circulating blood: the band form and the segmented
form (Fig. 8.9). The band form has a typical elongated
nucleus.
• A mature, segmented neutrophil has a characteristic
multilobed nucleus. The separate lobes are attached
to each other by a fine threadlike filament. The
filament, between separate lobes, may be hidden.
SEGMENTED NEUTROPHI
(FIG. 8.9) The nucleus of the basophil can be difficult to see
because it is usually obscured by dark, large
granules.
 • The band form of neutrophils, eosinophils,
and basophils and, in the final stage of
maturation, the segmented neutrophils,
eosinophils (Fig.8.10), and basophils (Fig.
8.11) are the cell forms normally found in the
circulating blood.

• Mast cells (tissue basophils) are not observed

in the blood of healthy persons. These cells
have an appearance similar to that of the
blood basophil. Mast cells have a round or
oval nucleus. The granules of the mast cell do
not overlie the nucleus as they do in basophils

FIGURE 8.10 EOSINOPHILIC SERIES FIGURE 8.11 BASOPHILIC

SERIES
 GRANULATION IN MATURE FORMS
THE CHARACTERISTICS OF THESE GRANULES ARE AS FOLLOWS:

• The granules of segmented neutrophils are rich in various antibacterial

substances, including lysosomal hydrolases, lysozyme, and myeloperoxidase.
Some of these granules are typical lysosomes.
• Eosinophilic granules - differ from neutrophilic granules in that they lack lysozyme.
These granules are of two types:
1. Smaller round granules, which have been identifi ed as not containing
crystalloids. These granules exist in small
quantities in the mature eosinophil and are rich in acid phosphatase.

2. Larger crystalline granules, which are more numerous. These crystalline

granules are elliptical, are larger than
the granules of the neutrophil, and have an amorphous matrix surrounding an
internal crystalline structure. The
crystals are thought to represent the enzyme peroxidase (not the same as the
myeloperoxidase found in neutrophils), and the matrix contains acid phosphatase.
• Basophilic granules contain heparin and histamine. Mast cells have
granules that have an enzyme content similar to those of the blood basophil.

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