GRANULOPEISIS

Sri Mulatsih
Subbagian Hematologi-onkologi,
IKA FK UGM/RSUP Dr. Sardjito
Yogyakarta
2008

Molecular mechanisms involved in

regulation of lineage choice
decisions during granulopoiesis

Hematopoiesis is a complex series of events which results in

the formation of all blood lineages.
The development of mature blood cells is regulated at various
levels:

self-renewal of pluripotent stem cells in the bone marrow,

proliferation of progenitors
and differentiation of hematopoietic stem cells to mature blood cells.

Aberrant regulation of hematopoiesis can lead to severe clinical

conditions ranging from myeloproliferative disorders to severe
immune deficiencies.
In order to develop novel therapies against illnesses caused by
aberrant regulation of hematopoiesis, it is necessary to
understand the molecular mechanisms underlying these
processes.

Cont

Cytokines, stromal cells and extracellular matrix proteins play an

important in regulation of hematopoiesis.
The PI3K/PKB signal transduction pathway plays a critical role in
regulation of lineage choice decisions during myeloid
development:
inhibition of PKB is required for eosinophil maturation, whereas
activation of PKB is necessary to induce neutrophil
differentiation.
Since PKB is regulated by both G-CSF and IL-5, additional
signals must be involved in down-regulation of PKB activity
during eosinophil differentiation.
Co-culture of hematopoietic progenitors with stromal cells
induces eosinophil but not neutrophil development, suggesting
that stromal cells might play an important role in inhibition of
PKB activity during eosinophil differentiation.

HEMATOPOEISIS

GRANULOPEISIS
Is

a complex process by which primitive

blood precursors differentiate into fully
differentiated, functionally active
granulocytes.
Maintains peripheral neutrophil counts in
the circulation at 1.8-9.0x109/L
Production in marrow: 1.6x109 cells/kg/d

TRANSCRIPTION FACTOR

Act both positively and negatively to regulate the

expression of a wide range of important genes:

growth factors and their receptors,

other transcription factors.

The activity of several transcription factors is

controlled by external stimuli, such as cytokines.
There is a complex interplay between all of these
factors - synergistic and antagonistic - which
allows for the exquisite control of granulocytic
cell production that is observed.

GRANULOPEISIS

Controlled by a myriad of transcription factors which

regulate the expression of essential genes.
Including those encoding growth factors and their
receptors, enzymes, adhesion molecules, and
transcription factors themselves.
In particular, C/EBP , PU.1, CBF, and c-Myb have
emerged as critical players during early granulopoiesis.
These transcription factors interact with one another as
well as other factors to regulate the expression of a
variety of genes important in granulocytic lineage
commitment.

GRANULOPOEISIS

GRANULOPOEISIS

GRANULOPOEISIS

GRANULOPEISIS

GRANULOPEISIS

GRANULOPEISIS

BONE MARROW

GRANULOPEISIS

MYELOBLAST

MYELOCYTE

METAMYELOCYTE

BANDS LEUKOCYTE

SEGMENTED NEUTROPHIL

EOSINOPHULIC LEUKOCYTE

BASOPHILIC GRANULOCYTE

EOSINOPHILIC LEUKOCYTE

G-CSF
Act

on both precursor cells and mature

terminally-differentiated neutrophil
Functions:

Stimulates the proliferation

Decreases apoptosis
Reduces the maturation times
Increasing the mature neutrophil by:

Augmenting the mitotic pool

Reducing the maturation time
Releasing the bone marrow storage pool

Granulopoeisis
In

peripheral:

6-12 hours
50% in axial stream
50% in marginal pool
Migrate into extravascular 1-2
day-apoptosis

Nutrophil

Segmented nucleus
Highly condensed
chromatin
Full complement
granules and
secretory vesicles

Neutrophil Granules and Secretory Vesicles

Four

type cytoplasmic inclusions:

Primary (azurophil) granulesMPO, protein

antimicrobial (BPI)
Secondary (specific) granules-lactoferrin
Tertiary (gelatinase) granules extravaculare
migration
Secretory vesicles

BPI
Functions:

Permeabilizing bacterial
membrane
Neutralizing endotoxin
Opsonizing bacteria
--phagocytosis

 Defensins

4 kD cationic protein
Cytotoxic activity:
Bacteria
Fungi
Parasites
Viruses

Lysozyme:

Lyzing bacteria wall (primary and secondary

granule)

Clinical correlation
Neutrophilia:

Increased mobilization neutrophil from the

bone marrow
Expansion of the progenitor pool or
shortening of the mitotic cycle
Failure to exit the circulation

Neutrophilia

Physiologic

Stimulus of exercise or epinephrine

Infection
Inflammations
Acute hemorrhage or hemolysis

Corticosteroid
Persistent inflammatory response
Leukemoid reaction
Post splenectomy

Genetic mutation 2 subunit CD 18 (LAD-1)

Genetic defect in selectin ligands

Acute response:

Chronic:

Disorder in neutrophil migration

Neutropenia
Decreased

production
Increased utilization
Peripheral destruction
Increased vascular margination

Neutropenia
Pseudo-neutropenia
Infection-Induced
Drug-Induced
Immune

Hemolytic anemia
Autoimmune disease
Chronic

Benign

Congenital neutropenia
Inhibitor

of CFU-GM
Or absence on local cytokine production

MIGRATION OF
GRANULOCYTE