WHITE BLOOD CELL

By
Dr. Trushika Patel,
Assistant Professor,
Department of Physiology,
Note : This is for the reference purpose only. You have to read your textbook. Kiran Medical College, Surat.
OBJECTIVES
• Introduction
• Classification
• Formation : Leukopoiesis
• Different WBCs with their functions and variation in count
Introduction
• The white blood cells (WBCs) or leukocytes are colorless in contrast
to the red colour of RBCs.

• These are nucleated cells and play an important role in the defence
mechanism of the body.

• The leukocytes of the peripheral blood are of two main varieties,

distinguished by the presence or absence of granules.

• These are granulocytes and agranulocytes (non-granulocytes).

Classification
Granulocytes

• The white blood cells with granules in their cytoplasm are called
granulocytes.

• Depending upon the colour of granules, granulocytes are further

divided into three types:
oNeutrophil
oBasophil
oEosinophil
Agranulocytes

• White blood cells which do not contain granules in their cytoplasm are
called agranulocytes.

• These are of two types:

o Lymphocytes and
o Monocytes.
Total leukocyte count

• Adults: 4000–11,000/μL of blood.

• At birth, in full-term infant: 10,000–25,000/μl of blood.

• Infants up to 1 year of age: 6000–16,000/μl of blood.

Differential leukocyte count (DLC)

Granulocytes
• Neutrophils : 40 -75 %
• Eosinophils : 1 - 6 %
• Basophils : 0 - 1 %

Agranulocytes
• Lymphocytes : 20 - 40 %
• Monocytes : 2 - 10 %
• The DLC determines if there is an increase or decrease in a particular
type of leukocyte, because in different diseases, one or the other type
of cells show an increase or decrease in its numbers.

• The differential count is done in 100 or 200 cells and shows only a
relative increase or decrease in particular variety of cells.
Variations In WBC Count
Leukocytosis
• Leukocytosis refers to increase in total WBC count above 11,000/μl.

Physiological causes of leukocytosis are:

1. Age,
2. Exercise,
3. After food intake,
4. Mental stress,
5. Pregnancy and
6. Exposure to low temperature.
Pathological causes of leukocytosis are:

1. Acute bacterial infections especially by the pyogenic organisms,

2. Acute haemorrhage,
3. Burns,
4. Post-operative period,
5. Tuberculosis and
6. Glandular fever.
Leukopenia
• Leukopenia refers to decrease in the total WBC count below 4000/μl.

Causes of leukopenia are:

• Infections by the non-pyogenic bacteria, especially typhoid fever and
paratyphoid fever.
• Viral infections, such as influenza, smallpox, mumps, etc
• Protozoal infections.
• Starvation and malnutrition.
• Aplasia of bone marrow.
• Bone marrow depression due to:
oDrugs, Repeated exposure to X-rays or radiations, Chemical poisons
like arsenic, dinitrophenol and antimony.
Leukaemia

• Leukaemias constitute a group of malignant diseases of the blood in

which there occurs an increase in the total WBC count associated with
presence of immature WBCs in the peripheral blood.

• The total WBC count is usually above 50,000/μ and may be as high as
100,000–300,000/μ.
Types of Leukaemias

1. Acute myeloblastic leukaemia,

2. Acute lymphoblastic leukaemia,
3. Chronic myeloid leukaemia and
4. Chronic lymphoid leukaemia.
LEUKOPOIESIS
• The process of development and maturation of white blood cells
(leukocytes), called leukopoiesis, is a part of haemopoiesis (formation
of blood cells).

• All the blood cells develop from the pluripotent haemopoietic stem
cells (PHSCs).

• The leukopoiesis can be discussed under two headings:

o Formation of granulocytes (granulopoiesis) and monocytes, and
o Formation of lymphocytes (lymphopoiesis)
 Leukopoiesis
 Myeloid stem cell  Lymphoid stem cell
oMyeloid series oLymphoid series
Formation of granulocytes
(granulopoiesis)
• Neutrophils Formation of lymphocytes
• Basophils (lymphopoiesis)
• Eosinophils • Small lymphocytes
• Large lymphocytes
oMonocyte –macrophage series
Formation of monocytes
Formation of Granulocytes
And
Monocytes
• The granulocytes and monocytes are formed in the bone marrow from
the colony forming unit called CFU-GM (colony forming unit
granulocytes and monocytes)

• The progenitor cells (CFU-GM) forming different cells are further

named as:

• CFU-G for neutrophil

• CFU-EO for eosinophil
• CFU-Ba for basophil
• CFU-M for monocyte.
• The development of granulocytes through various stages is called
myeloid series.

• The development of monocytes through various stages is called

monocyte–macrophage series.
Formation of Granulocytes

(Granulopoiesis)

Myeloid Series
 MYELOID SERIES
• The cells of myeloid series
include :

oMyeloblast (most primitive

precursor),
oPromyelocytes,
oMyelocytes,
oMetamyelocytes,
oBand forms and
oSegmented granulocyte (mature
form).
• The process of granulopoiesis takes about 12 days.

• Granulocytes are formed and stored in the bone marrow.

• When need arises they are released in circulation.

• Normally the number of granulocytes in bone marrow is about three

times as compared to circulating in the peripheral blood.
 Formation of Monocytes

Monocyte–Macrophage Series
 MONOCYTE–
MACROPHAGE SERIES

• Monocyte–macrophage series
include :

1. Monoblast (myelomonoblast)
2. Promonocyte
3. Monocyte
 MONOCYTE–
MACROPHAGE SERIES

1. Monoblast
(myelomonoblast):

• It is a large cell similar in structure

to the myeloblast.
2. Promonocyte:

• It is a young monocyte about 20 μm

in diameter.
• Its nucleus is large, indented (often
kidney shaped) and contains one
nucleolus.
• The nuclear chromatin is arranged in
a loose network.
• The cytoplasm is basophilic and
contains no azurophilic granules, but
may have fine granules which are
larger than those in the mature
monocyte.
3. Monocyte

• From the bone marrow, the

monocytes migrate into spleen and
lymphoid tissues.

• The transformed stages of these

cells in the various tissues are
called tissue macrophages and
form a part of tissue–macrophage
system, which was previously
known as reticuloendothelial
system.
Formation of Lymphocytes

Lymphoid Series
• The lymphocytes are formed from the lymphocyte stem cells which
are formed from the PHSCs in the bone marrow.

• The lymphocyte stem cells migrate into the thymus and the
peripheral lymphoid tissues, where they proliferate and mature into
lymphocytes.

• In man, the bone marrow and thymus form the primary

lymphopoietic organs, where lymphoid stem cells undergo
spontaneous division independent of antigenic stimulation.
• The tissues which actively produce lymphocytes from the germinal
centers of lymphoid follicles as a response to antigenic stimulation
constitute the so-called secondary or reactive lymphoid tissue.

• It is comprised of the:

o Lymph nodes
o Spleen
o Gut associated lymphoid tissue.
Lymphoid series
1. Lymphoblast

• It is the earliest recognizable cell of the lymphoid series.

• It is actively dividing cell and resembles the myeloblast

morphologically except for the following minor differences:

oNuclear chromatin is slightly clumped and stippled as compared to

the fine meshwork in myeloblast

oNuclear membrane is fairly dense as compared to very fine

membrane of the myeloblast.
2. Prolymphocyte

• It is the intermediate stage between the lymphoblast and mature

lymphocyte.

• Its features are:

o Diameter is 9–18 μm.
o Nucleus is round to indented with slightly stippled or coarse
chromatin and may have 0–1 nucleoli.
o Cytoplasm is scanty and non-granular
3. Lymphocytes

• Prolymphocytes mature successively into a large lymphocyte and a small

lymphocyte, both of which are found in circulation.

• Then some lymphocytes enter thymus where they are processed and come
out as T lymphocytes.

• In thymus, a factor called thymosin plays an important role in the

processing.

• Some lymphocytes are processed in liver (in fetal life) and bone marrow
(after birth). These come out as B lymphocytes.
Regulation of Leukopoiesis
Regulation of leukopoiesis
• During tissue injury and
inflammation, bacterial toxins,
products of injury, etc. cause a
great increase in the rate of
production and release of
leucocytes.

• Thus, the products of dead and

dying white cells themselves
control leukopoiesis.
• The cytokines which control the formation of different types of
granulocytes are called colony stimulating factors (CSFs).
oThe CSFs are glycoproteins formed by monocytes and T
lymphocytes.

• The cytokines which control the formation of lymphocytes are called

interleukins e.g. IL -1, IL-3, etc.
oThe interleukins are formed by monocytes, macrophages and
endothelial cells.

• Prostaglandins formed by monocytes and lactoferrin also control

leukopoiesis.
Different WBCs.
 Different WBCs
Granulocytes Agranulocytes

• Neutrophils • Monocytes
• Basophils • Lymphocytes
• Eosinophils oSmall lymphocytes
oLarge lymphocytes
 Neutrophil
• Constitution of WBC : 50-70%
• Diameter : 10-14 μm
• Cytoplasm : granular, Slight bluish
• Granules :
oVery fine like sand particles
oNumerous red brown or purplish in colour
• Nucleus : Multilobed (1-6 lobes), purple in colour
• Lifespan : 8 – 10 hrs
 Functions of neutrophils
• The neutrophils along with the monocytes constitute the first line of
defence against the micro-organisms, viruses and other injurious
agents that enter the body.

• Neutrophils play this role by the following mechanisms:

• Phagocytosis
• Reaction of inflammation
• Febrile response
Reaction of inflammation.
• The neutrophils release leukotriene, prostaglandins, thromboxane, etc.
that bring about the reactions of inflammation like vasodilatation and
oedema.

Febrile response.
• The neutrophils contain a fever producing substance called
endogenous pyrogen which is an important mediator of febrile
response to the bacterial pyrogens.
 Phagocytosis
• Phagocytosis (cell eating) refers to the process of engulfment and
destruction of solid particulate material by the cells.

• The neutrophils engulf the foreign particles or bacteria and digest them
and ultimately may kill them by a process called phagocytosis.
The process of phagocytosis involves following steps:

1. Margination
2. Emigration and diapedesis
3. Chemotaxis
4. Opsonization (attachment stage)
5. Engulfment stage
6. Secretion (degranulation) stage
7. Killing or degradation stage
A: margination; B: diapedesis; C: chemotaxis; D: opsonization; E: engulfment; F: formation of phagolysosome;
1. Margination
• In the area of infection, the neutrophils gets marginated.

• Neutrophils get attached towards the capillary endothelium and start

rolling along its surface. This process is called margination or
pavementing.

• It is caused by binding of selectins (cell adhesion molecules) present

on the endothelial cells with the carbohydrate molecules present on
the surface of neutrophils.
• The endothelial selectins are markedly increased in areas where there
is inflammation.
2. Emigration and diapedesis
• The marginated neutrophils are emigrated in large number from the
blood to the site of infection by diapedesis into the tissues.

• They pass through the junction between endothelial cells of the blood
vessels.
3. Chemotaxis
• Chemotaxis refers to the process by which the neutrophils are
attracted towards bacteria at the site of inflammation.

• The process of Chemotaxis is mediated by the chemotactic agents

called chemokines.

• They are released at the infected area.

• There are various types of chemokines e.g. Leukotriene B4, cytokines.

• The chemokines increase the adhesive nature of neutrophils which

form clumps surrounding the infected area.
4. Opsonization (attachment stage)
• Opsonization refers to the process of coating of bacteria by the
opsonins by which bacteria become tasty to the phagocytes.

• The principal opsonins are the naturally acting factors in the serum
and include IgG opsonin and opsonin fragment of the complement
protein.
5. Engulfment stage
• The neutrophils project pseudopodia in all directions around the
opsonized particle which is bound to the surface of neutrophil.
• Pseudopodia meet each other on opposite side and fuse.
• This creates an enclosed chamber with the engulfed material.
• It breaks away from the membrane forming a phagocytic vesicle.
• Then the lysosomes of the cell fuse with the phagocytic vesicle to
form phagolysosome or phagosome.
6. Secretion (degranulation) stage
• Once the bacterium is engulfed, the lysosomes pour their enzymes into
the vesicle and also in interstitial space. This process is called
degranulation.

• There are large numbers of proteolytic enzymes especially geared up

for digesting bacteria.

• In addition, lysosomes of macrophages also contain lipases which can

digest the thick lipid membranes possessed by certain bacteria.
7. Killing or degradation stage
• The neutrophils and macrophages contain bactericidal agents
(defensins α and β) which can kill most of the bacteria.

• The bactericidal substance accomplishes the killing process by the

following mechanisms:

• Oxygen-dependent bactericidal mechanism

• Oxygen-independent bactericidal mechanism
• Myeloperoxidase mechanism
• Phagocytosis is completed after the stage of killing is over.

• A neutrophil can usually phagocytose 5–20 bacteria before it itself

become inactivated or dead.

• Neutrophils are not capable of phagocytosing particles much larger

than bacteria.

• Neutrophils killed by the toxins released from the bacteria are

collected in the centre of infected area.
A: margination; B: diapedesis; C: chemotaxis; D: opsonization; E: engulfment; F: formation of phagolysosome;
 Variations In Counts
Neutrophilia

• Neutrophilia refers to increase in the circulating neutrophil counts

(absolute count > 10,000/μl).

• Commonest cause of leucocytosis.

 Causes of Neutrophilia
Physiological causes Pathological causes
• Newborn babies, • Acute pyogenic bacterial infections,
• After exercise, • Non-infective inflammatory
• After meals, conditions like gout, acute
rheumatic fever,
• Pregnancy, menstruation, parturition
and lactation, • Acute tissue destruction as in:
oBurns,
• Mental stress and emotional stress,
oPost-operatively and
• After injection of epinephrine. oMyocardial infarction.
Neutropenia
• Decrease in the neutrophil count is known as neutropenia (absolute
count < 2500/μL).

Causes
• Typhoid and paratyphoid fever,
• Malaria,
• Aplasia of bone marrow,
• Bone marrow depression due to:
oDrugs, such as chloromycetin and cytotoxic drugs used in the
malignant diseases
oRepeated exposure to X-rays and radiations
oChemical poisons like arsenic.
 Arneth count
• Counting the number of neutrophils with different nuclear lobes and
expressing the count as percentage of cells with different number of
nuclear lobes is called Arneth or Cooke’s Arneth count.

Clinical significance.
• The Arneth count is useful in judging the rate of formation of
neutrophils.
• The three-lobed cells are fully mature and functionally the most
efficient.
• Thus, presence of younger cells (shift to the left) and more mature
cells (shift to the right) in the blood can provide important information
about the rate of formation and release of neutrophils from the bone
marrow.
 Arneth count
Stage Nuclear lobes Diagram Normal Range
Stage 1 Nucleus is unilobed. 2-5%
(N1) The nucleus is C or U shaped.

Stage 2 Nucleus is bilobed. 20-30%

(N2) Two lobes are separated by thin strands.
Nucleus is trilobed.
Stage 3
Three lobes are separated by thin 40-50%
(N3)
strands.
Nucleus is tetralobed. 10-15%
Stage 4
Four lobes are separated by thin
(N4)
strands.
Stage 5 Nucleus is pentalobed. 2-5%
(N5) Five lobes are separated by thin strands.
 Eosinophil
• Constitution of WBC : 1-4%
• Diameter : 10-14 μm
• Cytoplasm : granular, Light pink,
• Granules : large and Brick red in colour,
• Nucleus : prominent and bilobed, purple in colour
• Lifespan : 8-12 days
• Function : It secretes an enzyme that modulate
allergic reaction and inflammatory reactions caused
by the foreign body.
 Functions of Eosinophils
Mild phagocytosis.
• Eosinophils are not very motile and thus have a very mild phagocytic
activity.

Role in parasitic infestations.

• They play an important role in the defence mechanism of body
especially in parasitic infestations.
Role in allergic reaction.
• The eosinophils increase in number in allergic conditions like
bronchial asthma and hay fever.

Role in immunity.
• The eosinophils are present in abundance in the mucosa of respiratory
tract, gastrointestinal tract and urinary tract, where they probably
provide mucosal immunity.
 Variations In Counts
Eosinophilia
• Eosinophilia refers to increase in the eosinophil count (absolute count
> 500/μL).

Causes
• Allergic conditions like bronchial asthma and hay fever,
• Parasitic infestation, e.g. intestinal worms like hookworm, roundworm
and tapeworm,
• Skin diseases like urticaria
• Scarlet fever.
Eosinopenia
• Eosinopenia is the decrease in the eosinophil count (absolute count
< 50/μL).

Causes
• Adrenocorticotrophic hormone (ACTH) and steroid therapy,
• Stressful conditions and
• Acute pyogenic infections.
 Basophil

• Constitution of WBC : 0-1%

• Diameter : 10-14 μm
• Cytoplasm : Blue & granular
• Granules : Purple or blue, Overlying the nucleus
• Nucleus : pale with bilobes or trilobes, purple in colour
• Lifespan : few hours to few days
• Function : It release histamine and heparin. Histamine
increases tissue blood flow. Heparin acts as an
anticoagulant.
 Functions of Basophils
Mild phagocytosis.
• Basophils have very mild phagocytic function.

Role in allergic reaction.

Role in preventing spread of allergic inflammatory process.
Release of heparin which prevents clotting of the blood.
 Variations In Counts
Basophilia
• Basophilia refers to increase in the basophil count (absolute count
> 100/μL).

Causes
• Viral infections, e.g. influenza, small pox and chicken pox,
• Allergic diseases and
• Chronic myeloid leukaemia.
Basopenia
• Decrease in the basophil count is called basopenia.

Causes
• Corticosteroid therapy,
• Drug-induced reactions and
• Acute pyogenic infections.
 Monocyte

• Constitution of WBC : 2-8%

• Diameter : 10-18 μm, Irregular outline
• Cytoplasm : agranular
• Granules : absent
• Nucleus : Single; usually kidney shape, Pale in colour,
Peripheral in position
• Lifespan : hours to days
• Function : differentiation into macrophages that
removes the dead cell and pathogens out of the spleen.
 Functions of Monocytes
Role in defence mechanism.
• Monocytes along with the neutrophils play an important role in the
body’s defence mechanism.

Role in tumor immunity.

• Monocytes may also kill tumor cells after sensitization by the
lymphocytes.

Synthesis of biological substances.

• Monocytes synthesize complement and other biologically important
substances.
 Variations In Counts
Monocytosis
• A rise in the blood monocytes above 800/μl is termed monocytosis.

Causes
• Certain bacterial infections, such as tuberculosis, syphilis and subacute
bacterial endocarditis.
• Infectious mononucleosis or the so-called glandular fever.
• Viral infections.
• Protozoal and rickettsial infections, e.g. malaria and kala-azar.
Monocytopenia
• Monocytopenia refers to decrease in the monocyte count.

Causes
• Monocytopenia is rare.
• It may be seen in the hypoplastic bone marrow
 Lymphocyte
• Constitution of WBC : 20-30%
• Diameter of Small lymphocyte : 7-10 μm
• Diameter of Large lymphocyte : 10-14 μm
• Cytoplasm : agranular, Scanty, Sky blue colour, less
than the amount of nucleus
• Granules : absent
• Nucleus : Single, Very big, occupying whole of the
cells, Purple in colour, Oval or rounded, Eccentric
Small lymphocyte Vs. Large lymphocyte
Small lymphocyte Large lymphocyte
• Nucleus occupies 80-90% of • Nucleus is indented, more
cytoplasm cytoplasm
 Functions of Lymphocytes
• Lymphocytes play an important role in immunity.

• B lymphocytes as well as their derivatives, the plasma cells are

responsible for the development of humoral immunity also called
antibody-mediated immunity.

• T lymphocytes are responsible for the development of cellular

immunity, also called cell-mediated immunity or T cell immunity.
 Variations In Counts
Lymphocytosis

• Lymphocytosis refers to increase in the lymphocyte count (absolute

count > 4000/μL).
 Causes of Lymphocytosis
Physiological causes Pathological causes

• In healthy infants and young • Chronic infections like tuberculosis,

children, the lymphocytes count hepatitis and whooping cough,
is usually high (about 60% in • Viral infections like chicken pox,
DLC) while the TLC is normal
(relative lymphocytosis). • Autoimmune diseases like
thyrotoxicosis,
• Infectious mononucleosis and
• In females, during menstruation
lymphocytes are increased • Lymphatic leukemia (most common
cause of lymphocytes > 10,000/μl).
Lymphopenia

• Lymphopenia or lymphocytopenia refers to decrease in lymphocyte

count (absolute count below 1500/μl).

Causes
• Patients on corticosteroid and immunosuppressive therapy,
• Hypoplastic bone marrow,
• Widespread irradiation and
• Acquired immunodeficiency syndrome
 Questions
• Classify the leucocytes and describe their morphological
features with the help of diagram.
• Elaborate the steps involved in the phagocytic function of
neutrophils.
• Describe the physiological roles of the different types of
granulocytes circulating in blood.
• Write down the variations in count of total WBCs with their
causes.
• Write down the variations in count of differential WBCs
with their causes.
• Functions of WBCs.
• Leukopoiesis
• Granulopoiesis
THANK YOU