Pre-Clinical Sciences 2

Hematology

Myelopoiesis and maturation of

leukocytes

Dr. Merin Chandanathil

Department of Physiology
mchandanathil@auamed.net
Office- GC31, EXT 1105
 Background knowledge
New concept in Reviewed concept Applicable clinical Connected material
current course from previous context from
courses other disciplines
Myelopoiesis PHY.8.4. Differentiate Significance of total Immunology-
and maturation white blood cells count and Immunological
based on their
of leukocytes microscopic
differential count in functions of WBCs
appearance and clinical practice, Pathology-
function. cytopenia abnormalities in
and cytoses, count and
change in morphology in
morphology of diseases
WBCs in diseases
HL1: Apply • HL.1.1 Outline the steps of hematopoiesis
knowledge of with emphasis on myelopoiesis.
physiology to • HL.1.2 Classify leukocytes based on their
morphology and function
understanding • HL.1.3 Analyze the process of leukopoiesis,
the development including the regulation of leukocyte
and maturation production in the bone marrow and the factors
influencing their differentiation and
of different types maturation.
of leukocytes and • HL.1.4 Evaluate the significance of differential
leukocyte counts in clinical practice
their clinical
significance
 A 55-year-old female presented to the emergency
department with fever and chest pain. Physical
examination indicated respiratory distress. A
chest X-ray showed an opacity in the left upper
lobe. Laboratory findings showed an elevated
Case study white blood cell count of 22,000/mm3, with a
differential count indicating 90%
polymorphonuclear leukocytes, 5% bands, 3%
lymphocytes, and 2% monocytes. These findings
raise concerns for an underlying condition that
requires further investigation to determine the
cause.
 White blood cells or leukocytes
Age Normal WBC count
• Five different types-neutrophils, lymphocytes,
monocytes, eosinophils, and basophils Newborns 9000-30,000 cells/mm3
• Normally, the quantity and morphology of WBCs Children age <2 years 6200-17,000 cells/mm3
are constant and predictable.
Adults and children age >2
• Infections and hematopoietic malignancies can 4000-11,000 cells/mm3
years
alter WBC levels and appearance significantly
• Total WBC count vs. Differential leukocyte count
• High total count or leukocytosis Cell type Percentage Absolute count
• Infectious diseases
• Steroid medications Neutrophils 57%-67% 2500-7400/mm3
• Inflammation Lymphocytes 25%-33% 1125-3630/mm3
• Low total count or leukopenia
• Bone marrow failure (due to malignancy, toxins, or
Monocytes 3%-7% 135-770/mm3
other unknown causes) Eosinophils 1%-3% 45-330/mm3
• Certain types of infection (such as HIV)
• Congenital diseases that impair WBC production. Basophils <1% <110/mm3
CLASSIFICATION-WHITE BLOOD
CORPUSCLES(WBC)

• Granulocytes
• Neutrophils
• Eosinophil
• Basophil
• Agranulocytes
• Lymphocytes
• Small lymphocytes
• Large lymphocytes
• Monocytes
 • The marrow produces around 200 billion red cells,
100 billion platelets, and 60 billion neutrophils
daily, matching the rate of peripheral destruction
• Hematopoiesis must persist throughout life
• It must be finely tuned and highly responsive to
changes in peripheral blood counts.
• Cytopenias (too few formed elements) and

Introduction
cytoses (too many formed elements) can have
serious, potentially fatal consequences
• The hematopoietic system relies on a hierarchy of
progenitor cells
• HSCs are at the top of the hierarchy and are
multipotent, giving rise to all hematopoietic cells
• HSCs have self-renewal ability, ensuring
maintenance of HSC numbers
 HSCs and progenitors
• HSC division
• Symmetric division
• Can lead to either two HSCs or two committed
progenitors
• With 2 HSC often occur in the fetal liver, increasing
HSC numbers
• Symmetric divisions leading to differentiation occur
after hematopoietic stress
• Asymmetric division
• One HSC and one committed progenitor, common in
the bone marrow
• Early progenitors may differentiate into
either myeloid or lymphoid lineages
• With further divisions, progenitors matures
and become restricted to single cell types
Differentiation and maturation of HSC
• Differentiation
• Cell becomes part of a particular cell line (eg: neutophilic
cell line)
• This depends on specific signaling molecule like cytokines
and growth factors
• Maturation
• The process of a cell moving through the stages of that cell
line
• From blast to mature blood cell (eg: Myeloblast to
neutrophil)
Hematopoiesis
Leukopoiesis
• WBCs arise from
• Myeloid cell line
• WBCs can develop in two
directions.
• Monoblasts develop into
monocytes
• Myeloblasts can develop into
neutrophils, eosinophils, and
basophils
• Lymphoid cell lines
• Lymphoblasts lead to B cells (B
lymphocytes), T cells (T
lymphocytes), and natural killer
cells.
 Granulopoiesis
• Formation of granulocytes- Neutrophil, Eosinophil
and Basophil
• Myeloblast and promyelocyte can differentiate into
any type of granulocyte
• Neutrophilic /Basophilic/ Eosinophilic Myelocytes
• Differentiation into early cells
• Granules specific to the cells are seen
• Neutrophilic /Basophilic/ Eosinophilic metamyelocyte
• Similar to previous stage with a more condensed
chromatin
• Band form
• Horse-shoe shaped nucleus
• Seen in circulation
• An increase may indicate the presence of inflammation or
infection
• Segmented or mature neutrophil
• Three or four small sections of the nucleus joined by thin
strings of filament or nuclear membrane
 Control of myelopoiesis by growth factors
• G-CSF (Granulocyte colony-stimulating
factor) is a key growth factor for
neutrophil progenitors
• Secreted by macrophages, endothelial
cells, and fibroblasts
• G-CSF production increases in response
to inflammatory cytokines (e.g., IL-1,
tumor necrosis factor)
• Stimulates granulocytic progenitors in the
marrow to increase neutrophil production
• G-CSF are glycoproteins that bind to
specific cytokine receptors which can
stimulate signaling through several
downstream pathways, including the
JAK/STAT, RAS, and AKT pathways
• G-CSF and GM-CSF is primarily used in
the treatment of drug-induced
neutropenia
 White blood cells differentials
Age Normal WBC count
• Differential can provide important Newborns 9000-30,000 cells/mm3
clues to the type of infection. Children age <2 years 6200-17,000 cells/mm3

• Increase in specific cells are Adults and children age >2

years
4000-11,000 cells/mm3
termed- neutrophilia,
lymphocytosis, monocytosis,
eosinophilia, or basophilia Cell type Percentage Absolute count
Neutrophils 57%-67% 2500-7400/mm3
• Decrease in cells is designated by Lymphocytes 25%-33% 1125-3630/mm3
the suffix -penia Monocytes 3%-7% 135-770/mm3
Eosinophils 1%-3% 45-330/mm3
Basophils <1% <110/mm3
 Neutrophil
• Neutrophils are about twice the size of
RBCs.
• Known as polymorphonuclear cells
due to nuclei with 3-5 lobes.
• Immature neutrophils are called band
cells with a single band-like nucleus.
• Infection and inflammatory state
causes change in morphology to
increase band cells called a left shift,
toxic granulation, cytoplasmic
vacuoles and Dohle bodies.
• Hypersegmented neutrophils (more
than 5 lobes) are seen in vitamin B12
and folate deficiencies (megaloblastic
anemia)
 Granules of neutrophils
• Neutrophil Granules
• Primary (Azurophilic) Granules
• Darker-stained dots.
• Contain myeloperoxidase, acid phosphatase, serine
proteases, beta-glucuronidase, and defensins.
• Comprise ~20% of the granules.
• Secondary (Specific) Granules
• Pink, smaller, and blend into the background.
• Contain leukocyte alkaline phosphatase, lysozyme,
collagenase, lactoferrin, and NADPH oxidase.
• Make up ~80% of the lysosomes.
• Tertiary Granules
• Contain gelatinase, cathepsins, and glycoproteins.
• Function of Neutrophils
• Phagocytosis and destruction of pathogens, necrotic
tissue, and foreign substances.
 Eosinophil
• Average diameter of 12 to 17 µm
• They have a bilobed nucleus
• Eosinophilic granules are large (0.5-1.5 µm)
and have a bright orange-red glow due to
high arginine content
• Eosinophil Functions
• Defend against parasitic infections
• Modulate some allergic responses (eg, in
asthma)
• Adrenal insufficiency
• Neoplasia
• Eosinophilic granules contain
• Major basic protein (defend against
helminthic infections)
• Eosinophilic cationic protein
• Histaminase
• Eosinophil peroxidase
• Eosinophil derived neurotoxin.
 Basophil
• Least common type of white blood cell,
comprising less than 1% of leukocytes
• The nucleus is usually bilobed but often
obscured by granules
• Large granules, which have an affinity
for basic dyes and stain dark blue to
purple
• The specific granules within both cells
contain histamine (a vasodilator)
and heparin (an anticoagulant).
• Basophilia is seen in
myeloproliferative disorders
 Monocyte- Macrophage
• Monocyte Development
• Start in bone marrow as monoblasts
• Promonocyte is the only stage between
monoblast and mature monocyte
• Mature monocytes have an indented,
irregular nucleus (kidney bean or
horseshoe-shaped)
• Characteristics
• Largest leukocyte
• Have gray-blue “dishwater” cytoplasm and
horseshoe-shaped nucleus
• Live 10-20 hours in blood, then migrate to
tissues and transform into macrophages or
dendritic cell
• Macrophages survive for months, serving as
the first line of defense
 Monocyte- Macrophage
• Functions of macrophages
• Phagocytize bacteria, viruses, and debris in
the tissues
• Recognize pathogens through pattern-
recognition receptors
• Recruit T-helper cells by secreting cytokines
• Serve as antigen-presenting cells to
enhance adaptive immunity
• Tissue-Specific Macrophages
• Kupffer cells in the liver
• Microglia in the brain
• Alveolar macrophages in the lungs
 Lymphocyte
• Found mainly in lymphatic organs (spleen, tonsils,
lymph nodes), some in blood.
• Lymphocyte Development:
• Start as lymphoblasts, prolymphocyte and then the
mature lymphocytes (small or large lymphocyte)
• Final differentiation gives unique appearances: Small
lymphocyte is T or B cell and large lymphocyte is NK cells
• Types of Lymphocytes:
• B cells: Adaptive immunity, humoral response, can
become antibody-secreting plasma cells.
• T cells: Adaptive immunity, cell-mediated response.
• NK cells: Innate immunity, large with granules, kill virus-
infected and tumor cells.
• Lymphocytosis Causes:
• Commonly due to viral infections (e.g., cytomegalovirus,
Epstein-Barr virus).
• Other causes: bacterial infections, leukemias,
lymphomas, drug hypersensitivity.
SUMMARY
Summary-Case study
A 55-year-old female presented to the emergency department with
fever and chest pain. Physical examination indicated respiratory
distress. A chest X-ray showed an opacity in the left upper lobe.
Laboratory findings showed an elevated white blood cell count of
22,000/mm3, with a differential count indicating 90%
polymorphonuclear leukocytes, 5% bands, 3% lymphocytes, and
2% monocytes. These findings raise concerns for an underlying
condition that requires further investigation to determine the cause.