Composition of Blood

• Consists of formed elements (cells)

suspended & carried in plasma (fluid
part)
• Total blood volume is about 5L
• Plasma is straw-colored liquid consisting
of H20 & dissolved solutes
– Includes ions, metabolites, hormones,
antibodies

13­7
Physiology of Blood
-Cells
By

Dr A. K. Gupta
MD (Pediatrics)
Ex . J.N. Medical College , A.M.U,
Aligarh
CMO (NFSG) ,Health Dept,GNCT
of Delhi
 Plasma Proteins

• Constitute 7-9% of plasma

• Three types of plasma proteins: albumins,
globulins, & fibrinogen
– Albumin accounts for 60-80%
• Creates colloid osmotic pressure that draws H20 from
interstitial fluid into capillaries to maintain blood volume &
pressure
• Globulins carry lipids
– Gamma globulins are antibodies
• Fibrinogen serves as clotting factor
– Converted to fibrin
– Serum is fluid left when blood clots
13­8
Red Blood Cells (Erythrocytes)
The major function of red blood cells :
Transport hemoglobin, which carries
oxygen from the lungs to the tissues.

Why Hb Should be inside RBCs:

When Hb is free in the plasma, 3 % of Hb
will leak through capillaries into tissue
spaces or glomerular filtrate each time the
blood passes through the capillaries.
So, Hb must be inside red blood cells to
remain in the human blood.
Red blood cells are responsible for
most of the acid-base buffering power
of whole blood? Explain
1.RBCs contain carbonic anhydrase
enzyme which catalyzes the reversible
reaction between carbon dioxide (CO2) and
water to form carbonic acid (H2CO3). This
reaction makes it possible for the water of
the blood to transport CO2 in the form of
bicarbonate ion (HCO3-) from the tissues to
the lungs, where it is reconverted to CO2
and expelled into the atmosphere as a
body waste product.
Shape and Size of Red Blood Cells.
Biconcave discs
Mean diameter -7.8 micrometers
The average volume of the red blood cell
is 90 to 95 cubic micrometers.
The shapes of red blood cells can
change as the cells squeeze through
capillaries. Because the normal RBC has
an excess of cell membrane for the
quantity of material inside, deformation
while passing through smallest cappilaries
does not affect the membrane,so the Red
Cells do not rupture passing through
Concentration of Red Blood Cells in
the Blood.
The average number of red blood cells is
Men- 5,200,000 (±300,000) per cumm
Women- 4,700,000 (±300,000) per cumm
Quantity of Haemoglobin in
the Cells & amount of O2
carried.
MAXIMUM CAPACITY : 34 Gm Hb in
100 ml of red cells.
Men: Hb: 15 grams per 100 ml of cells;
and
Women: Hb: 14 grams per 100 ml.
Each gram of pure haemoglobin
carries 1.34 ml of oxygen.
Therefore, in man 100 ml of blood can
 HEMOPOIESIS: INTRO
• Hemo: Referring to blood cells
• Poiesis: “The development or production of”
• The word Hemopoiesis refers to the
production & development of all the blood
cells:
– Erythrocytes: Erythropoiesis
– Leucocytes: Leucopoiesis
– Thrombocytes: Thrombopoiesis.
Production of Red Blood Cells Body

Areas of the Body That Produce Red

Blood Cells.
•In the early weeks of embryonic life- yolk
sac.
•During the middle trimester of
gestation: liver (main), spleen and lymph
nodes (additional sites).
•Then, during the last month of gestation
and after birth: exclusively in bone
marrow.-
•Till 5 yrs age: The bone marrow of
essentially all bones
•5 – 20 years age- The marrow of the long
bones, becomes fatty and produces no more
red blood cells after about age 20 years.
 Genesis of Blood Cells
Pluripotential Hematopoietic Stem
Cells, Growth Inducers, and
Differentiation Inducers.
The blood cells begin their lives in the bone
marrow from pluripotential hematopoietic
stem cell (PHSC).
There are successive divisions of the
pluripotential cells to form the different
circulating blood cells.
The intermediate-stage cells are very much
like the pluripotential stem cells, but are
committed to a particular line of cells and
 CLONAL HYPOTHESIS
PLURIPOTENT STEM CELL

MULTIPLICATION COMMITTMENT

COMMITTED
STEM CELL STEM CELL
MULTIPLICATION
COMMITTED
STEM CELL
PROGENITOR
CELL

CFU: COLONY
FORMING UNIT
CFU
A committed stem cell that produces
erythrocytes is called a colony-forming
unit-erythrocyte, and the abbreviation CFU-
E is used to designate this type of stem cell.
Colony-Forming Units which form granulocytes
and monocytes have the designation CFU-GM.
Growth Inducers:
Growth inducers are proteins that control the
growth and multiplication of the different stem
cells. Eg interleukin-3 , promotes growth
and reproduction of all the different types of
committed stem cells
Differentiation Inducers
Differentiation inducers are proteins which
causes differentiation of one type of
committed stem cell into a final adult blood
ERYTHROPOIESIS
 FACTORS REGULATING
ERYTHROPOIESIS
• SINGLE MOST IMPORTANT
REGULATOR: “TISSUE
OXYGENATION”
• BURST PROMOTING ACTIVITY
• ERYTHROPOIETIN
• IRON
• VITAMINS:
– Vitamin B12
– Folic Acid
• MISCELLANEOUS
 ERYTHROPOIETIN
• A hormone pro duced by the
Kidne y.
• A ci rc ul ati ng Gl yc oprote in
• Now adays availa bl e a s Syn theti c
Epoi eti n BY RE COMBI NANT
TECHNOL OGY
• Ac ts m ai nly o n CFU – E
• Inc rea se s the number of :
– Nucle ate d precur sor s in th e marr ow.
 VITAMINS
• B 12 : Cyanoc obal amine & F ol ic
Ac id:
– Is also cal led Ex tr insic F actor of
Cast le .
– Needs th e Intr insic Factor from th e
Gastri c j ui ce for ab so rpti on from
Smal l I nt est in e.
– De ficiency cause s Pe rnicious (Wh en
IF i s mi ssi ng) or Meg alo blasti c
An emia .
– Sti mul ate s Er yt hr opoie sis
 IRON
• Esse ntia l for the synthes is of
Hemo glo bin.
• Def icie ncy c ause s Mi croc yti c,
Hypo chromi c Ane mi a.
• The MCV ( ME AN CORP USCUL AR
VOL UME -NORMAL 90) , Co lo r
Inde x & MCH ( ME AN
CORP USCUL AR HE MOGL OBIN) a re
low .
 ERYTHROPOIESIS: SEQUENTIAL CHANGES
I II III IV
PV E
VI I L R M
R A N A E A
O R T T T T
N L E E I U
O Y R N C R
R N N B U E
M B B L L R
O L L A O B
B A A S C C
L S S T Y
A T T T
S E
T

MITOCHONDRIA
BASOPHILIA
HEMOGLOBIN
 ERYTHROID PROGENITOR CELLS
• BF U-E : Bu rst Fo rming Unit –
Erythroc yte :
– Give rise eac h to th ousan ds of
nuc leate d e ryth roid p re curso r cell s,
in vi tr o.
– Undergo some ch an ges to becom e
th e Co lon y F or ming U nit s-
Er yth rocyt e (CFU -E )
– Regul ato r: Bur st Pr om ot ing Ac tivi ty
(BPA )
 ERYTHROID PROGENITOR CELLS

• CFU- E: C olo ny Formi ng Uni t-

Erythroc yte :
– We ll d iff eren ti at ed eryt hroi d
proge ni tor cell.
– Pr ese nt on ly i n th e Re d Bon e
Mar row .
– Can form up to 64 nuc leate d
eryt hr oid precur sor cell s.
– Regul ato r: Er yth rop oi eti n.
• Bo th the se Pro ge nit or c ell s
cannot be dist ing uis hed ex ce pt
 Normoblastic Precursors

• PROE RY THROB LAST

(PRONORMOB LAST ):
– Lar ge ce ll : 15 – 20 Microns in
diam ete r.
– Cy to plasm is dee p v iolet- blue
stai ning
– Has no H emo gl ob in.
– Lar ge n ucle us 12 Mi crons o ccup ies
3/4 th of th e cell vol um e.
– Nucle us has fi ne sti ppled re ticulu m
& man y n ucle ol i.
 Normoblastic Precursors

• EARLY
NORMOBLAST(BASOPHILIC
ERYTHROBLAST):
– Smaller in size.
– Shows active Mitosis.
– No nucleoli in the nucleus.
– Fine chromatin network with few
condensation nodes found.
– Hemoglobin begins to form.
– Cytoplasm still Basophilic.
 Normoblastic Precursors

• INTERMEDIATE
NORMOBLAST(REYTHROBLAST):
– Has a diameter of 10 – 14 Microns.
– Shows active Mitosis.
– Increased Hemoglobin content in the
cytoplasm
– Cytoplasm is Polychromatophilic.
 Normoblastic Precursors

• LATE NORMOBLAST:
– Diameter is 7 – 10 Microns.
– Nucleus shrinks with condensed
chromatin.
– Appears like a “Cartwheel”
– Cytoplasm has a Eosinophilic
appearance.
 Normoblastic Precursors
• RETICULOCYTE:
– The penultimate stage cell.
– Has a fine network of reticulum like
a heavy wreath or as clumps of dots
– This is the remnant of the basophilic
cytoplasm, comprising RNA.
– In the Neonates, Count is 2 –
6/Cu.mm.
– Falls to <1 in the first week of life.
– Reticulocytosis is the first change
seen in patients treated with Vit B12
 Normoblastic Precursors
• MATURE ERYTHROCYTE:
– Biconcave disc.
– No nucleus.
– About One-third filled with
Hemoglobin.
Role of the Kidneys in Formation of
Erythropoietin.
1. 90 per cent of all erythropoietin is
formed in the kidneys; 10% in the
liver.
2. in the kidneys the erythropoietin is
formed probably in the renal tubular
epithelial cells which secrete the
erythropoietin,
3. when BOTH the kidneys are
destroyed by renal disease, the
person invariably becomes very
anemic because erythropoietin formed
Effect of Erythropoietin in
Erythrogenesis.
•Stimulate the production of
Proerythroblasts from hematopoietic
stem cells in the bone marrow.
•Speeds up Erythropoeisis by making
proerythroblasts rapdly pass through
different erythroblastic stages
• The erythropoietin mechanism for
controlling red blood cell production is
a powerful one.
•In the absence of erythropoietin, few
red blood cells are formed by the bone
marrow.
Maturation of Red Blood Cells-
Requirement for Vitamin B12 and Folic
Acid
•Vitamin B12 and folic acid are essential for
the synthesis of DNA as they form
thymidine tri-phosphate. Therefore, lack of
either of them cause
•Failure of rapid multiplication of
erythroblastic cells.
•Production of larger than normal red cells
called macrocytes which are fragile and
thus have a short life, one half to one third
normal.
Pernicious Anemia.

•In pernicious anemia there is red blood cell maturation

failure due failure to absorb vitamin B12 from the gastrointestinal
tract due to atrophic gastric mucosa.
•The parietal cells of the gastric glands secrete a glycoprotein called
intrinsic
factor.
•The Intrinsic factor binds tightly with the vitamin B12. In this bound
state, the B12 is protected from digestion by the gastrointestinal
secretions.
•In the bound state, intrinsic factor binds to specific receptor sites on
the brush border membranes of the mucosal cells in the ileum.
•Then, vitamin B12 is transported into the blood during the next few
hours by the process of pinocytosis, carrying intrinsic factor and the
vitamin together through the membrane.
• Lack of intrinsic factor, therefore, causes diminished bsorption of
the vitaminB12.
•Once vitamin B12 has been absorbed from the gastrointestinal tract,
it is first
stored in the liver, then released slowly as needed by the
bone marrow.
The minimum amount of vitamin B12 required each day to
maintain normal red cell maturation
Failure of Maturation Caused by
Deficiency of Folic Acid
(Pteroylglutamic Acid).
Folic acid is a normal constituent of green
vegetables, some fruits, and meats
(especially liver). It is easily destroyed
during cooking.

In small intestinal disease called sprue,

there is highly reduced absorbption of
both folic acid and vitamin B12
 Formation of Hemoglobin
•Synthesis of hemoglobin begins in the
proerythroblasts and continues even
into the reticulocyte stage of the red
blood cells. When reticulocytes leave the
bone marrow and pass into the blood
stream, they continue to form minute
quantities of hemoglobin for another day or
so until they become mature erythrocytes.
Types of haemoglobins
The different types of chains alpha
chains, beta chains, gamma chains, and
delta chains form diferent types of
haemoglobins:
1.HbA: The most common form of
hemoglobin in the adult human.
It is a combination of two alpha chains
and two beta chains.
It has a molecular weight of 64,458.
A total of four molecules of oxygen (or
eight oxygen atoms) that can be
transported by each hemoglobin
2. The types of hemoglobin chains in
the hemoglobin molecule determine
the binding affinity of the hemoglobin
for oxygen.
For instance, in sickle cell anemia, the
amino acid valine is substituted for
glutamic acid at one point in each of the
two beta chains.
When this type of hemoglobin is exposed
to low oxygen, it forms elongated
crystals inside the red blood cells
which make it almost impossible for
the cells to pass through small
Combination of Hemoglobin with
Oxygen.
1. Oxygen binds loosely with one of
the coordination bonds of the iron
atom. This is an extremely loose bond, so
that the combination is easily reversible.
2. Furthermore, the oxygen does not
become ionic oxygen but is carried as
molecular oxygen (composed of two
oxygen atoms) to the tissues, where,
because of the loose, readily reversible
combination, it is released into the tissue
fluids still in the form of molecular oxygen
Iron Metabolism
Uses of Iron in body:
1.For the formation of hemoglobin
2. For formation of other elements like e.g.,
myoglobin, cytochromes, cytochrome oxidase,
peroxidase, catalase

The total quantity of iron in the body averages

4 to 5 grams,
1. 65 % in the form of hemoglobin.
2. 4 per cent is in the form of myoglobin,
3. 1 per cent is in the form of the various
heme compounds that promote intracellular
oxidation,
4. 0.1 per cent is combined with the protein
transferrin in the blood plasma,
 Daily Loss of Iron.
1. Excretion in faeces: 0.6 milligram per day,
2. Loss due to bleeding in woman due to menstrual loss :1.3
mg/day.

Absorption of Iron from the Intestinal Tract

Iron is absorbed from all parts of the small intestine, mostly by
the following
mechanism.
•The liver secretes moderate amounts of apotransferrin
into the bile, which flows through the bile duct into the
duodenum.
•In dudenum the apotransferrin binds with iron and
withhemoglobin and myoglobin from meat. This combination is
called transferrin.
•Transferrin binds with receptors in the membranes of
the intestinal epithelial cells.
•Then, by pinocytosis, the transferrin molecule, carrying
its iron store, is absorbed into the epithelial cells and
released into the blood capillaries beneath these cells in the
form of plasma transferrin.
•Iron absorption from the intestines is very slow, only a
Regulation of Total Body Iron by
Controlling Rate of Absorption.
2.When the body has become saturated
with iron so that essentially all apoferritin
in the iron storage areas is already
combined with iron, the rate of additional
iron absorption from the intestinal tract
becomes greatly decreased.
3.On the contrary, when the iron stores
have become depleted, the rate of
absorption can increase five times normal.
Thus, total body iron is regulated by
altering the rate of absorption.
Life Span and Destruction of Red Blood Cells Body:
When red blood cells are delivered from the bone marrow into
the circulatory system, they normally circulate an average of
120 days before being destroyed.
Even though mature red cells do not have a nucleus,
mitochondria, or endoplasmic reticulum, they do have
cytoplasmic enzymes that are capable of metabolizing
glucose and forming small amounts of adenosine
triphosphate.
These enzymes also
(5)maintain pliability of the cell membrane,
(6)maintain membrane transport of ions,
(7)keep the iron of the cells' hemoglobin in the ferrous form
rather than ferric form, and
(8)prevent oxidation of the proteins in the red cells.

Once the red cell membrane becomes fragile many of the

red cells self-destruct in the spleen, where they squeeze
through the red pulp of the spleen. There, the spaces
between the structural trabeculae of the red pulp,
through which most of the cells must pass, are only 3
micrometers wide, in comparison with the 8-micrometer
diameter of the red cell.
 Destruction of Hemoglobin.
•When red blood cells burst and release their
hemoglobin, the hemoglobin is phagocytized
almost immediately by macrophages in many
parts of the body, but mainly Kupffer cells of
the liver and macrophages of the spleen and
bone marrow.
•The macrophages release iron from the
hemoglobin and pass it back into the blood, to
be carried by transferrin either to the bone
marrow for the production of new red blood
cells or to the liver and other tissues for
storage in the form of ferritin.
•The porphyrin portion of the hemoglobin
molecule is converted by the macrophages,
through a series of stages, into the bile
pigment bilirubin, which is released into the
blood and later removed from the body by
 Anemias
Anemia means deficiency of hemoglobin in the
blood, which can be caused by:
•Blood Loss Anemia.
•After rapid hemorrhage, the body replaces the
fluid portion of the plasma in 1 to 3 days, but this
leaves a low concentration of red blood cells. The
Hb returns to normal by 3 to 6 weeks.
•In chronic blood loss, a person frequently
cannot absorb enough iron from the intestines to
form hemoglobin as rapidly as it is lost. Red cells
are then produced that are much smaller than
normal and have too little hemoglobin inside them,
giving rise to microcytic, hypochromic anemia,
which is shown in Figure in next slide.
2. Aplastic Anemia. Bone marrow aplasia
means lack of
functioning bone marrow. For instance, a person
exposed to gamma ray radiation from a nuclear
3. Megaloblastic Anemia.
Deficiency of Vitamin B12, folic acid, and
absence of intrinsic factor from the
stomach mucosa lead to slow reproduction
of erythroblasts in the bone marrow.
As a result, the red cells grow too large,
with odd shapes, and are called
megaloblasts.
Thus, atrophy of the stomach mucosa, as
occurs in pernicious anemia, or loss of
the entire stomach after surgical total
gastrectomy can lead to megaloblastic
anemia.
Also, patients who have intestinal sprue, in
4. Hemolytic Anemia. Due to Breakdown
of RBCs.
Inherited defects of Red cell membrane
make the cells fragile, so that they rupture
easily as they go through the capillaries,
especially through the spleen.
The number of red blood cells formed
may be normal in hemolytic diseases but
the life span of the fragile red cell is so
short that the cells are destroyed faster
than they can be formed.
Eg
a. hereditary spherocytosis- the red cells
are very small and spherical rather than being
biconcave discs. These cells cannot withstand
compression forces because they do not have
the normal loose, baglike cell membrane
b. Sickle cell anemia-
The RBCs have an abnormal type of hemoglobin
called hemoglobin S, containing faulty beta chains in
the hemoglobin molecule,
When this hemoglobin is exposed to low
concentrations of oxygen, it precipitates into
long crystals inside the red blood cell.
These crystals elongate the cell and give it the
appearance of a sickle rather than a biconcave
disc. The precipitated hemoglobin also damages the
cell membrane, so that the cells become highly
fragile, leading to serious anemia.
Such patients frequently experience a circle of events
called a sickle cell disease “sicke cell crisis," in which
low oxygen tension in the tissues causes sickling, which
leads to ruptured red cells, which causes a further
decrease in oxygen tension and still more sickling and
red cell destruction.
c. Erythroblastosis fetalis, Rh-positive red blood cells
in the fetus are attacked by antibodies from an Rh-
negative mother. These antibodies make the Rh-
Effects of Anemia on Function of the
Circulatory System.
•The blood viscosity falls.
•This decreases the resistance to blood flow in
the peripheral blood vessels, so that far
greater than normal quantities of blood flow
through the tissues and return to the heart,
thereby greatly increasing cardiac output.
•Moreover, hypoxia resulting from
diminished transport of oxygen by the
blood causes the peripheral tissue blood
vessels to dilate, allowing a further
increase in the return of blood to the
heart and increasing the cardiac output
to a still higher level-sometimes three to four
times normal.
•Thus, one of the major effects of anemia is
5. When a person with anemia begins to
exercise, the heart is not capable of
pumping much greater quantities of blood
than it is already pumping. Consequently,
during exercise, which greatly
increases tissue demand for oxygen,
extreme tissue hypoxia results, and
acute cardiac failure ensues
 Polycythemia
Secondary Polycythemia (Production of
large number of RBCs) It occurs when
there is hypoxia in tissues because of too
little oxygen in the breathed air, such as at
high altitudes, or because of failure of oxygen
delivery to the tissues, such as in cardiac
failure, so the erythropoeitin stiumuate blood-
forming organs to automatically produce large
quantities of extra red blood cells.
This condition is called secondary
polycythemia, and the RBCs count rises to
6 - 7 million/mm3 ie 30 per cent above normal.
Eg physiologic polycythemia, occurs in
Polycythemia Vera (Erythremia). In addition to
those people who have physiologic polycythemia,
others have a pathological condition known as
polycythemia vera, in which the red blood cell
count may be 7 to 8 million/mm3 and the
hematocrit may be 60 to 70 per cent instead of the
normal 40 to 45 per cent. Polycythemia vera is
caused by a genetic aberration in the
hemocytoblastic cells that produce the blood cells.
The blast cells no longer stop producing red cells
when too many cells are already present. This
causes excess production of red blood cells in the
same manner that a breast tumor causes excess
production of a specific type of breast cell. It
usually causes excess production of white blood
cells and platelets as well.
In polycythemia vera, not only does the hematocrit
increase, but the total blood volume also increases,
on some occasions to almost twice normal. As a
result, the entire vascular system becomes
Effect of Polycythemia on Function of the
Circulatory System
•Because of the greatly increased
viscosity of the blood in polycythemia,
blood flow through the peripheral blood
vessels is often very sluggish.
•The color of the skin depends to a great
extent on the quantity of blood in the skin
subpapillary venous plexus. In polycythemia
vera, the quantity of blood in this plexus is
greatly increased. Further, because the blood
passes slowly through the skin capillaries
before entering the venous plexus, a larger
than normal quantity of hemoglobin is
deoxygenated. The blue color of all this
deoxygenated hemoglobin masks the red
 Leukocytes (White Blood Cells)
The leukocytes, also called white blood
cells,
They are formed partially in the bone
marrow (granulocytes and monocytes and
a few lymphocytes) and
partially in the lymph tissue
(lymphocytes and plasma cells).
After formation, they are transported in the
blood to different parts of the body where
they are needed in areas of serious
infection and inflammation, thereby
providing a rapid and potent defence
Genesis of leukocytes
Types of White Blood Cells.
Six types of white blood cells are normally
present in the blood. They are
1. Polymorphonuclear neutrophils, 2.
Polymorphonuclear eosinophils,
3. Polymorphonuclear basophils, 4. Monocytes, 5.
Lymphocytes, and, 6. occasionally, Plasma cells.
The first three types of cells, the
polymorphonuclear cells, all have a granular
appearance, for which reason they are
called granulocytes, or, in clinical
terminology, "polys," because of the
multiple nuclei.
The granulocytes and monocytes protect
the body against invading organisms mainly
by ingesting them-that is, by phagocytosis.
Concentrations of the Different White
Blood Cells in the Blood.
Normal WBC count: Total Leukocyte
Count (TLC) =7000 per ml of blood (in
comparison with 5 million red blood cells).
The normal % of the different types
called Differential Leukocyte Count
(DLC) is :
Polymorphonuclear neutrophils 62.0%
Polymorphonuclear eosinophils 2.3%
Polymorphonuclear basophils 0.4%
Monocytes 5.3%
Lymphocytes 30.0%
 INTRODUCTION
• Leucocytes: Mobile units of the
body’s defence mechanism
• Formed in the :
– Bone marrow
– Lymphoid tissue.
• Rapidly deployed through the
blood to areas where:
– Infection &
– Inflammation are seen.
 CLASSIFICATION OF LEUCOCYTES
LEUCOCYTES
100

GRANULOCYTES AGRANULOCYTES

EOSINOPHILS BASOPHILS MONOCYTES LYMPHOCYTES

2.3 0.4 5.3 30

NEUTROPHILS
62
 NEUTROPHILS
• Most numerous Leucocytes ( 50 –
70%)
• Are 10 – 14 Microns in diameter.
• Have a constantly changing
shape due to amoeboid
movements.
• The Nucleus can have 1 – 7 lobes
connected by a fine strand.
• The Cytoplasm contains 50 – 200
fine granules.
 NEUTROPHILIA
• NEUTROPHILIA: Increased
neutrophil count, can be due to:
– Release of stored cells from the
bone marrow reserves.
– Bacterial Infections causing
increased Neutropoiesis.
– Exercise can cause release of
stored neutrophils.
 Cytoplasmic Granules
• Fine, azurophilic (Stain with both
Eosin & Methylene blue) in
nature.
• Contain enzymes such as:
– Cathepsins.
– Phosphatases.
– Nucleases.
• Granules serve as lysosomes.
 NEUTROPHILS & MONOCYTES:
Functions
• They seek, attack and destroy
invading bacteria, viruses and other
injurious agents
• Neutropils attack and destroy
bacteria and viruses, even in the
blood.
• Monocytes are immature until they
enter the tissues. There, they swell
up to 80 Microns, develop lysosomes,
and become Macrophages, capable of
defence.
Neutrophils & Macrophages
• Diapedesis: They squeeze
through the pores of the
blood vessels.
• Amoeboid movement: They
move at rates several times
their own length!
• Chemotaxis: Directed
movement – cells move to
wards infected areas.
 EOSINOPHILS
• 3 – 8% of the
Leucocytes.
• Have a typical
‘Spectacle
shaped’, bilobed
nucleus.
• Have coarse
bright pink
staining granules
in the cytoplasm.
 EOSINOPHILS

• They are parsiticidal in function.

• Eosinophilia or increased count
occurs in:
– Parasitic infestations.
– Allergic conditions.
– TPE: Tropical Pulmonary
Eosinophila.
 Basophils
• Are very few in number: <
1%.
• Have a large indented
nucleus which is obscured
by cytoplasmic granules.
• Granules are coarse and
basic staining: blue.
• They are abundant and
protrude through the cell
membrane.
LYMPHOCYTES: IMMUNOCYTES
• Morphologically,
– LARGE
Lymphocytes:
– Sized about 12 –
15 µ
– Thin cytoplasmic
rim
– Large spherical
nucleus
– No cytoplasmic
granules.
LYMPHOCYTES:
IMMUNOCYTES
Small
Lymphocytes:
– Sized about 8 µ.
– ( Smallest
Leucocytes)
– Thin cytoplasmic
rim & Large
spherical
nucleus.
– No granules
visible.
 LYMPHOCYTES:
IMMUNOCYTES
• Physiological Classification: T
and B
– ‘T’ LYMPHOCYTES :
• Thymus trained or schooled
cells
• Responsible for Cell
mediated immunity.
• Provide protection against
intracellular pathogens
LYMPHOCYTES: IMMUNOCYTES:
‘B’ LYMPHOCYTES
– Trained in the Bone
marrow(Bursa Fabricius in
birds)
– Responsible for Humoral
Immunity.(Immunity through
Antibody production)
– Protect the body from
encapsulated pyogenic
bacteria like Pneumococcus &
streptococci.
 LEUCOCYTES: REVIEW
• Chemotaxis: Directed movement
• Chemotaxins: Cytotaxins &
Cytotaxigens
• Eosinophils: Spectacle shaped
nucleus, Coarse pink granules.
• Basophils: Coarse blue granules,
obscuring nucleus. Custard apple
appearance.
• Lymphocytes: Physiologically T and B
types.Responsible for immunity:
Immunocytes.
 Life Span of the White Blood Cells
The life of the granulocytes after being released from
the bone marrow is normally 4 to 8 hours circulating in
the blood and another 4 to 5 days in tissues where they
are needed.
The monocytes :
•Have a short transit time, 10 to 20 hours in the blood, before
wandering through the capillary membranes into the tissues.
•Once in the tissues, monocytes swell to larger sizes to become
tissue macrophages, and, in this form, can live for months
unless destroyed while performing phagocytic functions.
•These tissue macrophages are the basis of the tissue
macrophage system, which provides continuing defense
against infection.
Lymphocytes :
8.Enter the circulatory system continually, along with drainage
of lymph from the lymph nodes and other lymphoid tissue.
9.After a few hours, they pass out of the blood back into the
tissues by diapedesis. Then, still later, they re-enter the lymph
and return to the blood again and again; thus, there is
continual circulation of lymphocytes through the body.
10.The lymphocytes have life spans of weeks or months; this
life span depends on the body's need for these cells.
3. Neutrophils and Macrophages
Defend Against Infections
It is mainly the neutrophils and tissue
macrophages that attack and destroy
invading bacteria, viruses, and other
injurious agents. The neutrophils are
mature cells that can attack and destroy
bacteria even in the circulating blood.
How White Blood Cells Enter the
Tissue Spaces:
1. By Diapedesis. Neutrophils and
monocytes can squeeze through the pores
of the blood capillaries by diapedesis.
2. by Ameboid Motion. Both neutrophils
and macrophages can move through the
tissues by ameboid motion.
3. Attracted to Inflamed Tissue by
Chemotaxis. Many chemical substances in
the tissues cause both neutrophils and
macrophages to move toward the source of
the chemical. This phenomenon is known
How White Blood Cells Enter the
Tissue Spaces
 Phagocytosis
A. The most important function of the neutrophils and
macrophages is phagocytosis, which means cellular ingestion
of the offending agent. Phagocytes must be selective of the
material that is phagocytized; otherwise, normal cells and
structures of the body might be ingested.
B. Whether phagocytosis will occur depends especially
on three selective procedures.:
•First most natural structures in the tissues have
smooth surfaces, which resist phagocytosis. But if the surface
is rough, the likelihood of phagocytosis is increased.
•Second, most natural substances of the body have
protective protein coats that repel the phagocytes.
Conversely, most dead tissues and foreign particles have no
protective coats, which makes them subject to phagocytosis.
•Third, the immune system of the body develops
antibodies against infectious agents such as bacteria. The
antibodies then adhere to the bacterial membranes and
thereby make the bacteria especially susceptible to
phagocytosis.
C. The Complement 3 molecules attach to receptors on the
phagocyte
membrane, thus initiating phagocytosis. This selection and
D. Mechanism of Phagocytosis by
Neutrophils.
1.On approaching a particle to be
phagocytized, the neutrophil first attaches
itself to the particle and then projects
pseudopodia in all directions around the
particle.
2.The pseudopodia meet one another on the
opposite side and fuse. This creates an
enclosed chamber that contains the
phagocytized particle.
3.Then the chamber invaginates to the inside
of the cytoplasmic cavity and breaks away
from the outer cell membrane to form a free-
E. Phagocytosis by Macrophages.
•Macrophages are the end-stage product of
monocytes that enter the tissues from the blood.
•They are much more powerful phagocytes
than neutrophils, often capable of phagocytizing
as many as 100 bacteria.
• They have the ability to engulf much larger
particles,
•Also, after digesting particles, macrophages
can extrude the residual products and often
survive and function for many more months.
•Once Phagocytized, Most Particles Are
Digested by Intracellular ENZYMES.
Both neutrophils and macrophages contain an
abundance of lysosomes filled with proteolytic
enzymes especially geared for digesting bacteria
and other foreign protein matter. The lysosomes of
macrophages (but not of neutrophils) also contain
large amounts of lipases, which digest the thick
F. Both Neutrophils and Macrophages
Can Kill Bacteria.
•In addition to the digestion of ingested
bacteria,neutrophils and macrophages
contain bactericidal agents that kill
most bacteria even when the
lysosomal enzymes fail to digest
them.
•Much of the killing effect is due to
powerful oxidizing agents formed by
enzymes in the membrane of the the
peroxisome. These oxidizing agents
include large quantities of superoxide (O2-),
Acute Increase in Number of Neutrophils
in the Blood-"Neutrophilia."
Also within a few hours after the onset of
acute, severe inflammation, the number of
neutrophils in the blood sometimes increases
fourfold to fivefold-from a normal of 4000 to
5000 to 15,000 to 25,000 neutrophils per
microliter.
This is called neutrophilia, which means an
increase in the number of neutrophils in the
blood.
Neutrophilia is caused by products of
inflammation that enter the blood stream, are
transported to the bone marrow, and there act
on the stored neutrophils of the marrow to
mobilize these into the circulating blood. This
Eosinophils
The eosinophils normally constitute about 2 per cent of all
the blood leukocytes.
Eosinophils are weak phagocytes, and they exhibit
chemotaxis, but in comparison with the neutrophils, it is
doubtful that the eosinophils are significant in protecting
against the usual types of infection.
Eosinophils, however, are often produced in large
numbers in people with parasitic infections, and they
migrate in large numbers into tissues diseased by
parasites.
Although most parasites are too large to be phagocytized by
eosinophils or any other phagocytic cells, eosinophils attach
themselves to the parasites by way of special surface
molecules and release substances that kill many of the
parasites. For instance, one of the most widespread infections
is schistosomiasis,
Eosinophils attach themselves to the juvenile forms of the
parasite and kill many of them. They do so in several ways:
(1) by releasing hydrolytic enzymes from their granules, which
are modified lysosomes; (2) probably by also releasing highly
reactive forms of oxygen that are especially lethal to parasites;
and (3) by releasing from the granules a highly larvacidal
polypeptide called major basic protein.
Eosinophils also have a special
propensity to collect in tissues in which
allergic reactions occur, such as :
1.in the peribronchial tissues of the lungs in
people with asthma and in the skin after
allergic skin reactions.
2. The eosinophils are believed to detoxify
some of the inflammation-inducing
substances released by the mast cells and
basophils and probably also to phagocytize
and destroy allergen-antibody complexes,
thus preventing excess spread of the local
inflammatory process.
Basophils
The basophils in the circulating blood are similar to the large
tissue mast cells located immediately outside many of the
capillaries in the body.
Both mast cells and basophils liberate heparin into the
blood, a substance that can prevent blood coagulation.
The mast cells and basophils also release histamine, as well
as smaller quantities of bradykinin and serotonin. I
ndeed, it is mainly the mast cells in inflamed tissues that
release these substances during inflammation.
The mast cells and basophils play an exceedingly
important role in some types of allergic reactions
because the type of antibody that causes allergic reactions,
the immunoglobulin E (IgE) type has a special propensity to
become attached to mast cells and basophils.
Then, when the specific antigen for the specific IgE antibody
subsequently reacts with the antibody, the resulting
attachment of antigen to antibody causes the mast cell or
basophil to rupture and release exceedingly large quantities of
histamine, bradykinin, serotonin, heparin, slow-reacting
substance of anaphylaxis, and a number of lysosomal enzymes.
These cause local vascular and tissue reactions that cause
many, if not most, of the allergic manifestations.
Leukopenia (decreased WBC count)
A clinical condition known as leukopenia
occasionally occurs in which the bone
marrow produces very few white blood
cells, leaving the body unprotected
against many bacteria and other agents
that might invade the tissues.
Causes :
4.Irradiation of the body by x-rays or gamma
rays, or exposure to drugs and chemicals that
contain benzene or anthracene nuclei, is likely
to cause aplasia of the bone marrow.
5.Some common drugs, such as
chloramphenicol (an antibiotic), thiouracil
(used to treat thyrotoxicosis), and even
 The Leukemias
Uncontrolled production of white blood cells
can be caused by cancerous mutation of a
myelogenous or lymphogenous cell. This causes
leukemia, which is usually characterized by
greatly increased numbers of abnormal white
blood cells in the circulating blood.
Types of Leukemia.
Leukemias are divided into two general types:
•lymphocytic leukemias and 2. myelogenous
leukemias.
The lymphocytic leukemias are caused by
cancerous production of lymphoid cells,
usually beginning in a lymph node or other
lymphocytic tissue and spreading to other areas of
the body.
The Myelogenous leukemia, begins by
cancerous production of young myelogenous
cells in the bone marrow and then spreads
 Effects of Leukemia on the Body
2.The first effect of leukemia is metastatic
growth of leukemic cells in abnormal areas of
the body.
•Leukemic cells from the bone marrow
invade the surrounding bone, causing
pain and a tendency for bones to fracture
easily.
•Almost all leukemias eventually spread
to the spleen, lymph nodes, liver, and
other vascular regions, regardless of
whether the origin of the leukemia is in the
bone marrow or the lymph nodes.
•Common effects in leukemia are the
THROMBOCYTES:
PLATELETS
Characteristics of Platelets
1. Diameter: Platelets are non nucleated
minute discs of 1 to 4 micrometers
diameter & donot multiply.
2. Formed in the bone marrow from
fragmentation of Megakaryocytes under
action of Thrombopoeitin formed in
Liver.
3. The normal platelets count in the
blood is between 150,000 and 300,000
per ml.
4. Active factors in Cytoplasm of
platelets:
• Contractile Proteins actin , myosin
molecules, and thrombosthenin,
which help platelets to contract;
• Endoplasmic reticulum and the
Golgi apparatus that synthesize
various enzymes and store calcium ions;
• Mitochondria capable of forming
adenosine triphosphate (ATP) and
adenosine diphosphate (ADP); ADP
helps in aggregation of platelets.
• Fibrin-stabilizing factor-An important
protein
• A growth factor that helps repair
5. The cell membrane of the platelets
A glycoproteins coat which do not adhere
to normal endothelium but adhere to
injured areas of the vessel wall to the
injured endothelial cells & or any exposed
collagen from deep within the vessel wall.
It contains phospholipids that activate
multiple stages of the blood-clotting
process.
6. Half-life of platelets in the blood is 8 to
12 days.
 THROMBOCYTES: INTRO
• Normally 1.5 - 4.0 Lakhs/Cu.mm
in blood.
• Are 2 – 4 µ in diameter; smallest
blood cells.
• Developed from giant cells
called Megakaryocytes.
 THROMBOCYTES: STRUCTURE
• Spherical, oval, or rod-shaped colorless
bodies.
• Diameter is between 2 to 4 μ.
• When unstimulated, under Electron
Microscopy they appear as:
– Flattened discs
– Having a cell membrane
– And a Cytoplasmic matrix.
– Microtubules encircle the thrombocyte just below
it’s surface membrane.
 THROMBOCYTES:
STRUCTURE
• Do not have nuclei.
• Cannot reproduce.
• But behave functionally as whole cells.
• Cytoplasm includes active proteins
such as:
–Actin.
–Myosin.
–Thrombesthenin.
 THROMBOCYTES:
STRUCTURE
• Cell Organelles in thrombocytes
include:
– Lysosomal granules.
– Dense bodies: about 50 – 100 in number.
– Mitochondria & Enzyme systems which
produce:
• ATP
• ADP
– Enzyme systems producing:
• Prostaglandins – Local hormones.
 THROMBOCYTES:
Structures within
– Fine Glycogen granules.
– Microvesicles.
– Microtubules.
– Filaments.
– Granules:
• Dense: Serotonin, ADP etc.
• ά-granules: Clotting factors such as:
– Fibrin Stabilizing Factor (FSF) Factor
XIII
– PDGF: Platelet Derived Growth Factor
 THROMBOCYTES:
STRUCTURE
• Internal Membranous systems:
– Open Canalicular System:
• Spongelike invaginations
• Provide multiple channels for:
– Taking up Calcium ions
– Secreting granule contents.
– Dense Tubular System:
• Channels of S.E.R.
• Serves as an intracellular store for Calcium
ions.
IDENTIFY THROMBOCYTES!
 Platelet Physiology
• Have a half life of • About half of them
8 – 12 days. are removed by the
• Eliminated from Macrophages in the
circulation by the Spleen.
Tissue Macrophage • Platelet surface
system. membrane has
• Thrombocytes are Phospho lipids,
active structures. Cholesterol &
glycolipids
 THROMBOCYTES: FUNCTIONS
• Formation of Platelet plugs in
Hemostasis.
– Activation.
– Adhesion.
– Aggregation/ Accumulation.
– Cohesion or Plug formation
• Supporting Coagulatory mechanisms.
• Phagocytosis.
• Storage & transport of substances.
 PLATELET PLUG FORMATION
• It can by itself stop blood loss, if
the rent is small.
• Many such minute ruptures occur
thousands of times every day in
minute blood vessels.
• Platelets manage to plug these very
well, all by themselves.
 APPLIED ASPECTS
• Thrombocytopenia:
– Decrease in Thrombocyte count.(Normal:
1.5 – 4 Lakhs/cu.mm of blood)
– Critical Thrombocyte Count is
40,000/cu.mm.
– Causes Purpura:
• Multiple subcutaneous purplish blotches.
IDIOPATHIC THROMBOCYTOPENIC
PURPURA
– Usually cause not known.
– Called as Idiopathic Thrombocytopenic Purpura.
– Diagnosis by:
• Easy bruisability & Purpura.
• Critical Platelet count.
• Prolonged Bleeding Time.
– Can be treated by giving multiple transfusions
of:
• Whole blood
• Platelet rich plasma: PRP
 APPLIED ASPECTS (Contd.)
• With normal thrombocyte count,
purpura may occur in
• Thrombesthenic Purpura, where the
thrombesthenin is defective.
• Thrombocytosis can cause increased
predisposition for Thrombotic events.
PURPURA