OBJECTIVES

1. Describe the properties and composition of blood

2. Understand composition of plasma and their function

3. Identify characteristics and functions of the formed elements of

the blood.

4. Identify the chemical regulators of blood cell production and

describe the process of erythropoiesis.

5. Describe the ABO system of red blood cell antigens and explain
the significance of the blood types.

6. Delineate the process of hemostasis that restricts blood loss and

how it is ultimately destroyed.
 INTRODUCTION
Blood is a dynamic and complex connective tissue in fluid form.

It is the ‘fluid of life’

It carries oxygen from lungs to tissues and carbon dioxide to

the lungs.

It is known as ‘fluid of growth’

It carries nutritive substances and hormones to all the tissues.

It is the ‘fluid of health’

It protects the body against the diseases

Gets rid of the waste products and unwanted

 Properties Of Blood

1. Color
Is red in color because it contains more oxygen (Arterial blood)

Purple red because of more carbon dioxide (venous blood)

2. Volume
In a normal adult ……. 5.6 L

In a newborn baby …… 450 ml

In females, it is slightly less and is about 4.5 L

Accounts 8 % of body weight in a normal young healthy adult.

(Cont…) Properties of blood
3. pH
 The pH of blood is 7.35–7.45; x = 7.4 (alkaline)

4. Density (Specific gravity)

 Refers to the weight of blood compared to that of water
 Specific gravity of H2O is taken as 1g/ml (i.e., 1ml of H2O
weighing 1gm at 40C).

Male: 1.052 - 1.063

Female: 1.050 - 1.058

(i.e., 1ml of whole blood weighing 1.052 to 1.061 gm)

 (Cont…) Properties of blood

5. Viscosity (thickness)
 4 – 5 times more viscous than water (where water = 1)

Due to red blood cells and plasma proteins

6. Osmolarity = 300 mOsm or 0.3Osm

 This value reflects the concentration of solutes in the plasma.

7. Salinity = 0.85%
 Reflects the concentration of NaCl in the blood

8.Temperature is 38C, slightly higher than “normal” body

temperature.
 Composition Of Blood
Blood makes up one-fourth of the extracellular fluid.

It is the circulating portion of extracellular fluid

Two major components

1. Plasma (55%)
 Protein-rich fluid, within which cellular elements are suspended.

2. Cellular elements (formed elements 45%)

1. Red blood cells or erythrocytes
2. White blood cells or leukocytes
3. Platelets or thrombocytes.
 Fragments of megakaryocytes in marrow.
 Composition Of Blood (cont,..)

Cell count and volume

Centrifug
e

Plasma (55% of whole

blood)
Buffy coat:
leukocyte, platelets
(<1% of whole
blood) Formed
Erythrocytes (45%)
of whole blood element
Take a sample of whole s
blood in a test tube
Hematocrit (PCV) : The percentage of erythrocytes in the total blood
volume.
Normal PCV: In males = 40% to 45%
In females = 38% to 42%
Composition Of Blood (cont,..)
Blood Plasma
Plasma is a straw-colored clear liquid part of blood
 Water = 92%
 Protein = 7%
 The remaining 1% is
 Dissolved organic molecules (glucose, amino acids, lipids,
and vitamins)
 Ions (Na+, K+, Cl-, H+, Ca++, HCO3-) … Na+ and Cl- are the

most abundant electrolytes (ions) and trace elements

 Non-protein nitrogenous substances – lactic acid, urea,
creatinine and bilirubin
 Dissolved O2 and CO2 and hormone
Cont,…

FIG. Composition of Blood plasma

 Composition of Blood (cont,..)

Serum

Clear a yellowish fluid that oozes from blood clot.

Volume of the serum is almost the same as that of plasma (55%).

It is different from plasma only by the absence of fibrinogen

because it is converted into fibrin during blood clotting.

For clinical investigations, it is separated from blood cells and

clotting elements by centrifuging.

Doesn’t require anticoagulant chemicals (heparin, EDTA,
oxalate etc.)
Composition Of Blood (cont,..)

Plasma volume:

Number of regulatory mechanisms in the body maintain

homeostasis of plasma volume.

Osmoreceptors in the hypothalamus

Antidiuretic hormone (ADH) … posterior pituitary

Renin-angiotensin-aldosterone system (RAAS)

Cont,…

Figure 2: Regulation of blood pressure.

 Plasma Proteins
 Most of plasma proteins are synthesized by the liver, with the exception
of some globulins, known as immunoglobulins (antibodies) which are
synthesized by lymphocytes.
 They are the largest of the plasma constituents and dispersed as a colloid

 Do not exit through the narrow pores in the capillary walls

 Establish an osmotic gradient

(colloid osmotic pressure ) between blood and interstitial fluid.

 Force preventing excessive loss of plasma from the capillaries into the

interstitial fluid.
 Helps to maintain plasma volume
 Cont,…
There are three groups of plasma
protein
A. Albumin (60%-80% or 4.5gm/dl)
 The most abundant plasma
proteins

B. Globulins (40%, or 2.5 g/dl)

 Are three subclasses (alpha, beta

and gamma globulin)

C. Fibrinogen (4%, or 0.3 g/dl)

Is the key factor in blood
clotting
 Characteristics Of Plasma Proteins
Albumin - Is formed in the liver
 Provide the colloidal osmotic pressure (oncotic pressure) needed to

draw water from the surrounding tissue fluid into the capillaries.
 25 millimeters of mercury (mm Hg).
 Contributes about 80% of the total colloid osmotic pressure of the
plasma
 To maintain blood volume and pressure

 During malnourished conditions (deficiency of protein intake), blood

volume decreases due to decreased albumin and this effect results in

edema development.
 Albumin also serves in transporting nutrients (FFA, hormones, bilirubin

etc.)
 Cont,..
Globulin- three subtypes: (alpha1&2, beta, gamma)

Alpha and beta globulins are produced by the liver

 Contributes about 20% of the total colloid osmotic pressure of the

plasma
 Are highly specific to which passenger they will bind and carry

 Function in transporting lipids and fat-soluble vitamins, thyroid

hormone and iron.

Gamma globulins are antibodies produced by lymphocytes.
 Function in immunity
Fibrinogen: Is synthesized by the liver
 It is mainly involved in blood clotting
 Functions Of Blood
1. Transportation
 Respiratory – transport O2 from alveoli of lungs to different tissues and

CO2 from tissues to alveoli for elimination in the

exhaled air.
 Nutritive – transport glucose, amino acids, lipids and vitamins to different

parts of the body for growth and production of energy.

 Excretory – carries waste products (such as urea), excess water and ions to

the kidney for excretion in the urine.

2. Regulation
 Temperature – thermoregulatory mechanism in the body, i.e. the
balance between heat loss and heat gain in the body.
 Hormonal – carries hormones from their site of origin to target
tissues ….. regulatory functions
 Cont,…

Regulation of acid-base balance

 Plasma proteins and hemoglobin act as buffers and help in the regulation of acid-
base balance
3. Storage
 Blood serves as a readymade source for substances (protein, glucose, Na +, K+ )
 These substances are taken from blood during the conditions like starvation, fluid
loss, electrolyte loss, etc.
4. Protection (Defensive)
Clotting – The clotting mechanism protects against blood loss when vessels
are damaged (broken) by activating plasma protein and platelets.
Immune – By activating WBC to defend the body against foreign invaders.
• Synthesizing and utilizing antibodies
 Erythrocytes
Erythrocytes Structure
• Are the most abundant cell type of the cellular (formed) elements

 Are flattened, biconcave disc shaped cells, indented in the middle on

both

sides much like doughnut shape.

 Biconcave shapes provides large surface area (30% more surface

area) for diffusion of O2 across the membrane.

Normal value
 RBC count ranges between 4.8 and 5.4 million/cu mm of blood.

 5.4 x 106 (±300,000)/cu mm …. male

 4.7 x 106 (±300,000)/cu mm ….. female
Cont,…

7.8 - 8µm

Figure3: Dimensions of RBC.

Cont,…

FIG. Osmotic changes to red blood cell shape

 Cont,…
Normal size
7.8 - 8 µm in diameter, 2.2 µm thick (at the periphery), and 1 µm
( at the center) because of the biconcave shape.
Thinness of the cell enables O2 to diffuse rapidly between the
exterior and inner most regions of the cell.
 Surface area : 20 sq µ.
 Volume : 90 to 95 cu µ.
 RBCs membrane has a special type of cytoskeleton, which is made

up of actin and spectrin.

 Spectrin give erythrocytes their flexibility, that allows RBC cells

to change shape as they squeeze through the capillaries of

circulation
 Cont,…
 Are non-nucleated, no centrioles, no organelles (mitochondria,ER)

RBCs are simple membranous “bags” filled with hemoglobin

(Hgb) - 97% of cell contents and enzymes

 They generate ATP anaerobically and prevents O2 consumption
being
transported.
Erythrocyte enzymes
 Only a few non renewable enzymes remain within a mature
erythrocyte
Glycolytic enzymes
 For generating energy needed to fuel the active transport mechanisms
To maintain proper ion concentrations within the cell
 Cont,…
 Carbonic anhydrase
 It is critical in CO2 transport in the form of bicarbonate ion
(HCO3-) from the tissues to the lungs.

Properties
Rouleaux formation
- When the RBCS are observed under microscope, they pile up one above
another like the pile of coins.
Suspension stability
- During circulation, the RBCs remain suspended uniformly in the blood.
Specific gravity
- Specific gravity of RBC is 1.092 to 1.101.
Packed cell volume (It is 45% of whole the blood)
 The proportion of blood occupied by RBCs expressed in percentage.
Cont,…

Figure 4: Rouleau formation

 Erythropoiesis
 Erythropoiesis is the process of the production, development and
maturation of erythrocytes.
Site Of Erythropoiesis
A. In fetal life
Occurs in three stages:
1. Mesoblastic Stage – During 1st two months of intrauterine life, the RBCs
are produced from mesenchyme of yolk sac.
2. Hepatic Stage – From 4-6th month of pregnancy (Liver, Lymph nodes
and Spleen) produce RBC
3. Myeloid Stage – During the last three months of intrauterine life and
RBCs are above red bone marrow and liver.
Note: The bone marrow of all bones produces red blood cells until a
person is 5 years old.
 Cont,…
 Up to the age of 20 years: RBCs are

produced from red bone marrow of

all bones (long bones and all the flat
bones).
 After the age of 20 years: RBCs are

produced from membranous bones

like ribs, sternum, vertebrae, and
pelvic bones, but not in long bones
Figure 5: Relative rate of red like femur or tibia (because of fat
blood cells production in the bone
deposition) .
marrow of different bones at
 The marrow becomes less
different ages
productive as age increases.
 Genesis Of Erythropoiesis
Stem Cells
 The primary cells capable of self-renewal and differentiating into

specialized cells.
 Pluripotential hemopoietic stem cells are the primitive cells in the

bone marrow, which give rise to all types of blood cells.

Types Of Stem Cells

1. Embryonic stem cells

 Derived from the inner cell mass of a blastocyst

 Self-renewal capacity and pluripotent nature

 Remain in the body as adult stem cells

 Cont,…
2. Adult stem cells
 Undifferentiated multipotent progenitor cells found (children and
adults).
 Capable of dividing and reforming the dying cells and regenerating the
damaged tissues.
 Are found in the bone marrow
1. Hemopoietic stem cells, which gives rise to blood cells
2. Bone marrow stromal cells, differentiate into cardiac and skeletal
muscle cells
Cont,…

Hemopoietic stem cells

 Are uncommitted pluripotent hemopoietic stem cells (PHSC)

(in the bone marrow).

 When the cells are designed to form a particular type of blood

cell, are called committed PHSCs.

Is restricted to give rise to one group of blood cells

Are of two types

1. Lymphoid stem cells (LSC), …… lymphocytes and natural

killer (NK) cells
 Cont,…
2. Colony forming blastocytes, give rise to myeloid cells
 When grown in culture, will produce colonies of specific types of
blood cells.
 Colony-forming unit–erythrocytes (CFU-E) are stem cells that

produce RBC’s
 Colony-forming unit-granulocyte/monocyte (CFU-GM) gives rise

to granulocye and monocyte

 Colony forming unit-megakaryocytes (CFU-M) give rise to
Platelets
 Cont,…

 Growth and reproduction of the different stem cells are controlled

by protein called growth inducer. E.g. interleukin-3

 Formation of the growth inducers and differentiation inducers

is itself controlled by factors outside the bone marrow.

Exposure of the blood to low oxygen cause growth and

differentiation of RBC
Infectious diseases cause growth and differentiation of specific

type of WBC
Figure: Hematopoeisis
 Stages Of Differentiation Of Blood Cells

1. Proerythroblast

2. Early normoblast

3.Intermediate
normoblast

4. Late normoblast

5. Reticulocyte

6. Matured erythrocyte
 Cont,…
1.Proerythroblasts: Nucleus present and Hb synthesis begins
2.Basophile erythroblast: cells continue dividing with the very little
hemoglobin
3. Poly-chromatophil erythroblast: b/c of presence of Hb the
cytoplasm stains with both acidic as well as basic stains
4.Orthochromatic erythroblast : nucleus disintegrates and disappears,
the process is called pyknos, but quantity of Hb increases
5.Reticulocytes: No nucleus, but remnants of ER, mitochondria and
Golgi apparatus are present that are later extruded and the cells pass
from the bone marrow into the blood capillaries by diapedesis
6. Erythrocytes: Reticular network disappears have no nucleus, but Hb
forms and accounts to 97% of the cell volume) attains the biconcave
shape
Note: The above stages requires 7 days for the development and
maturation of RBC.
Stages Of Differentiation Of Blood Cells
Table : Important events during
erythropoiesis
 Regulation of Erythropoiesis
Erythropoiesis is hormonally controlled and depends on adequate
supplies of iron, amino acids, and vitamins (B, C, D, etc.,)

The total mass of RBCs in the circulatory system is regulated within

narrow limits, so
 An adequate number of RBCs is always available to transport O2

-------tissues
 The cells do not become so numerous that they impede blood flow

Any condition that causes the quantity of oxygen transported to the

tissues to decrease normally increases the rate of RBCs production.
 Cont,…
Hormonal controls (Erythropoietin)
 The principal stimulus for RBC production in low oxygen states in the
tissues (hypoxia)
 Hypoxial stimulates production of a transcription factor called hypoxia
inducible factor 1 (HIC-1) which,
 Stimulate erythropoietin (EPO) gene to increase EPO synthesis

 90 % of it is formed in the kidneys and the remainder is formed mainly

in the liver.
 Norepinephrine, epinephrine and prostaglandins stimulate erythropoietin
production.
Is triggered by:
1. Less RBCs from bleeding (hemorrhage) …….low blood volume
2. Less RBCs due to excess RBC destruction ….. low hemoglobin
3. Low oxygen levels (high altitude, pulmonary disease)
 Cont,…
 In an atmosphere of low oxygen erythropoietin begins to be formed

within minutes to hours, and it reaches maximum 24 hours.

 Then EPO causes rapid production of RBCs and speed up to pass

more quickly through different erythroblastic stages than they

normally do.
After 5 days of the onset of erythropoietin, new RBCs appear in

the circulating blood.

Production of RBCs continues until enough red blood cells have

been produced to carry adequate amounts of O2 to the tissues.

Cont,…

Figure Control of erythropoiesis

Figure : Reflex by which decreased oxygen delivery to the kidneys
increases erythrocyte production via increased erythropoietin
 Cont,…
Maturation factors
 Erythropoietic cells of the bone marrow are among the most rapidly

growing and reproducing cells in the entire body.

 Their maturation and rate of production are affected greatly by a

person’s nutritional status.

 For the final maturation of RBCs two important vitamins are

essentially required, Vitamin B12 and folic acid.

Vitamin B12 (Cyanocobalamin) and folic acid

Essential for DNA synthesis in early mitotic divisions leading to

the formation of erythrocytes

Cont,…
Vitamin B12 (Cyanocobalamin)

 Vitamin B12 is called extrinsic factor since it is obtained mostly

from diet and absorbed from gastrointestinal tract

 Its absorption from intestine requires the presence of intrinsic

factor of Castle.
 Intrinsic factor is a glycoprotein, which is secreted by parietal

cells of gastric gland

 Combines with vitamin B12 in food and makes the B12

available for absorption by the gut.

Cont,…
Mechanism of Intrinsic factor to bind vitamin B12:

I. Intrinsic factor binds tightly with B12

II. Intrinsic factor binds to specific receptor sites on mucosal cells

in the ileum

III. Vitamin B12 is transported in to blood by process of pinocytosis

IV. Stored in large quantities in the liver

The minimum amount of vitamin B12 required each day to

maintain normal red cell maturation is only 1 to 3 micrograms
Cont,…
Folic Acid (Pteroylglutamic Acid)
 Folic acid is also required for the synthesis of DNA and cell

maturation.
 Folic acid is a normal constituent of green vegetables, some

fruits, and meats (especially liver).

 In the absence of folic acid, the synthesis of DNA decreases

causing failure of maturation.

 This leads to anemia in which the cells are larger and appear in

megaloblastic (proerythroblastic) stage.

 Formation of Hemoglobin
 Hemoglobin (Hb) is the iron containing pigment (coloring)

matter of red blood cell (RBC).

 Is O2 binding protein of RBCs and found only in RBCs.

 It is a tetramer comprising four globular protein chains (globins)

Each centered around an iron-containing heme group

 Synthesis of hemoglobin begins in the proerythroblasts stage of

RBCs and continue until RBCs become mature erythrocytes.

 Its synthesis requires an adequate supply of iron in the diet (red

meat, beans, spinach, etc.).

Cont,…
 Heme portion of hemoglobin is synthesized in mitochondria

where as the protein part, globin is synthesized in ribosomes.

 Succinyl-CoA binds with glycine to form a pyrrole molecule.

 Four pyrroles combine to form protoporphyrin ix, which then

combines with iron to form the heme molecule.

 Each heme molecule combines with a long polypeptide chain
and forming a subunit of hemoglobin called a hemoglobin
chain
 Cont,…
 There are four types of polypeptide
chains namely, alpha, beta, gamma
and delta chains.
 Each of these chains differs from
others by the amino acid sequence.
 Each globin molecule is formed by
the combination of 2 pairs of chains
(2α and 2γ or 2α and 2β chains)
 sAdult hemoglobin contain two
alpha chains and two beta chains.
 Each polypeptide chain combines
with one heme molecule.
 Each hemoglobin molecule contains
4
polypeptide chains and 4 heme
 Cont,…
 A heme prosthetic group containing an atom of iron and in each

hemoglobin molecule there are four iron atoms

 Each of the iron atom can loosely bind with one molecule of O 2

 O2 does not combine with the two positive bonds of iron in the

hemoglobin molecule.
 Instead it binds loosely by coordination bonds of the iron atom,

so O2 is carried as molecular oxygen rather than ionic one

 Cont,…
Types of normal hemoglobin

Hemoglobin is of two types:

1. Adult hemoglobin (HbA)

2. Fetal hemoglobin (HbF)

 Structure of HbF is similar with HbA except that the β chains are

replaced by γ chains; that is, HbF is α2γ2

 Replacement of fetal hemoglobin by adult hemoglobin starts immediately

after birth and completed at10th - 12th weeks after birth

 The main difference b/n HbA and HbF is that HbF O2 content at a given

Po2 is greater than HbA

 B/c HbF binds 2,3-biphospho glycerate (2,3-BPG) less avidly
 Cont,…
Normal hemoglobin content
 Red blood cells have the ability to concentrate hemoglobin up to about

34 gm in each 100 ml of cells.

 Average hemoglobin (Hb) content in whole blood ranges from 14 to 16

g/dL per 100ml of cells.

 This value varies depending upon the age and sex of the individual.

 At birth : 25 g/dL because of increased number of RBCs

 After3rd month : 20 g/dL

 After 1 year : 17 g/dL

 From puberty onwards : 14 to 16 g/dL

 Cont,…
Sex
 In adult males : 15 g/dL per 100ml of cells

 In adult females : 14 g/dL per 100ml of cells

 Each gram of hemoglobin is capable of combining with 1.34 ml of

oxygen.
 In a normal man, about 20ml of O2 and in a normal woman 19ml of O2

can be carried in combination with hemoglobin in each 100 ml of

blood.
 The binding affinity of the hemoglobin molecules for oxygen is

determined by the type of hemoglobin chain.

 Cont,…
 Some anemias results due to abnormal hemoglobin chain that alters

physical characteristics of the hemoglobin molecule.

 For instance, sickle cell anemia

 Which is genetic defect in which glutamete (glutamic acid) in beta

chain of hemoglobin is replaced by valine

 Exposed to low oxygen, it forms elongated crystals inside the RBCs

 Form rigid chains that make the RBC stiff and unnaturally shaped,

like a crescent or sickle

 Impede the passage of RBCs through the small capillary

 Results logjam blocks blood flow through small vessels

Iron Metabolism
Iron
 Necessary for the formation the hemoglobin essential to carry

oxygen
 The total quantity of iron in the body averages 4 to 5 grams

 65 % of which is in the form of hemoglobin

 15-30% Fe stored in the liver and spleen in the form of ferritin

 4% is stored in the muscle as myoglobin

 1% is found in intracellular oxidative heme compound

Cont,..
 0.1% is found in the plasma as transferrin
Note: Copper (Cu2+ ),Cobalt (Co), Nickel, Vitamin C are essential
in Hb synthesis
 Dietary iron exists in two forms:

i. Heme Iron - iron is bound as part of a heme group found in

hemoglobin, available in fish, meat and chicken
ii. Non-heme Iron - inorganic iron, exists in the oxidized ferric iron
(Fe3+) form and is available in vegetables, grains and cereals
The daily dietary iron requirement:
10 mg/day ……..for males
15mg/day ……. for women and
30 mg/day ……..for pregnant women
 Cont,…
Absorption of Iron
 Iron is absorbed mainly from the small intestine through the

intestinal cells (enterocytes) by pinocytosis

 Almost all iron absorption occurs in the duodenum.

Two main steps are involved in absorption of iron into blood:

1. Absorption of iron from the lumen into small intestine epithelial

cells and

2. Absorption of iron from the epithelial cells into the blood

 The extent to which ingested iron is taken up by the epithelial cells

depends on the type of iron consumed.

Cont,…
a. If the ingested iron is dietary heme, it is absorbed more efficiently
than inorganic iron b/c the reduced ferrous iron.

b. If the ingested iron is dietary Fe3+, it requires presences of other

substances that aid its reduction to Fe2+ prior to absorption.
Gastric secretions …… dissolve the iron

Vitamin C increases (ascorbic acid)

Ferric reductase
 Heme iron and Fe2+ are absorbed across the luminal membrane by
separate energy-dependent carriers.
Heme is transported into the enterocyte by a separate heme
transporter (HT)
Fe2+ is carried in via divalent metal transporter 1(DMT1)
 Cont,…

Figure : Absorption of iron. HT – heme transporter, DMT1 – Divalent metal

transporter1, FP – Transporter ferroportin, Hp – Hesphestin, TF – Transferin,
HO2 – Heme oxygenase
Cont,…
 After absorption into the small-intestine epithelial cells, iron has two
possible fates:

1. Iron needed immediately for production of red blood cells is absorbed

into the blood via a membrane iron transporter known as ferroportin.
 It immediately combines with a β-globulin called apotransferrin

resulting in the formation of transferrin

 So, iron is transported in blood in the form of transferrin

 The normal plasma iron level is about 130 µg/dL (23 mol/L) in men

and 110 µg/dL (19 µmol/L) in women

 Iron is loosely bound in the transferrin and can easily released to any

tissue cells
 Cont,…
 Transferrin molecule binds strongly with receptors in the cell

membranes of erythroblasts
 It is then ingested into the erythroblasts by endocytosis and delivers

the iron to the mitochondria, where heme is synthesized.

2. Iron not immediately needed is stored within the small-intestine

epithelial cells
 Excess iron in the blood is deposited especially in the liver hepatocytes

 In the cell cytoplasm, iron combines mainly with a protein, apoferritin,

to form ferritin and iron stored as ferritin is called storage iron.

 Smaller quantities of the iron in the storage pool are in an insoluble
form called hemosiderin.
Cont,…
Decrease in plasma iron
causes, release of some of
the iron in storage pool in
the form of transferrin in
plasma to the area of the
where it is needed
People with inadequate
quantities of transferrin in
their blood, develop
hypochromic anemia (less
hemoglobin than normal)
Cont,…
Daily Loss Of Iron
 About 0.6 mg Fe is excreted in male everyday in the stools

 Women have a larger iron loss averaging 1.3mg/day because of the

additional iron lost during menstruation.

 One gram of hemoglobin contains 3.34 mg of iron

Regulation of total iron in the body

1. Iron absorption is largely controlled by a hormone, hepcidin ….
iron
 It prevent iron export by binding with ferroportin from small intestine

2. Stoppage of apotransferrin formation in the liver

3. Reduction in the release of iron from the transferrin

Life Span and RBC Destruction
 The average lifespan of RBC is about 120 days.

 Membrane becomes fragile, the cell ruptures and release their

hemoglobin.
 The hemoglobin is phagocytized by phagocytic cells in the liver,

spleen, and bone marrow.

Hemoglobin split into its components:
 Heme (Iron, porphyrin ) and Globin (to protein depot for later use)
 Iron is transported in plasma by beta-globulins (transferrin)
either to the bone marrow or to the liver for storage in the form
of ferritin.
 Porphyrin is degraded into bilirubin
Cont,…
Cont,…
Erythrocyte Disorders
 Anemia – a decrease in the ability of the blood to carry oxygen due to

i. A decrease in the total number of erythrocytes

ii. A diminished concentration of hemoglobin per erythrocyte; or

iii. A combination of both.

Causes of anemia:

Nutritional anemia - is caused by a dietary deficiency of a factor needed

for erythropoiesis. For example, iron deficiency anemia

Pernicious anemia - is caused by an inability to absorb enough ingested

vitamin B12 from the digestive tract.
-The problem is a deficiency of intrinsic factor
Cont,…
Aplastic anemia - Is caused by failure of the bone marrow to
produce enough RBCs
 Destruction of red bone marrow (radiation, benzen, Vira infections,
invasion of the marrow by cancer cells, or chemotherapy for cancer)
Renal anemia - Result from kidney disease. Because EPO is the
primary stimulus for promoting erythropoiesis
Hemorrhagic anemia - Is caused by losing a lot of blood.
 The Loss can be either acute, such as a bleeding wound, or chronic,
such as excessive menstrual flow
Hemolytic anemia - Is caused by the rupture of too many circulating
erythrocytes.
 External factors : invasion of RBCs by malaria parasites or
 Genetic defective, as in sickle cell disease and erythroblastosis fetalis
 Cont,…
Polycethemia:
Is characterized by too many circulating RBCs and an elevated hematocrit
There are two general types of polycythemia

A. Physiological (secondary polycethemia) - Is erythropoietin-induced to

improve the blood’s O2 carrying capacity in response to a prolonged

reduction in O2 delivery to the tissues

 The marrow produces excessive RBCs (6-7 million / mm3)
 It occurs normally in people living at high altitudes, where less O 2
or
 In people for whom O2 delivery to the tissues is impaired by
chronic lung disease or cardiac failure.
 Cont,…
B. Pathological (Polycethemia Vera (erythremia) or primary
polycythemia.
 Is caused by a tumor like condition of the bone marrow

 It is about 7-8 million / mm3 and the hematocrit may be 60 to

70%
 Increases blood’s viscosity up to 5-7 times normal causing

the blood to flow very sluggishly ….. reduce O2 delivery to

the tissues
 Moreover, increases are also seen in WBC and platelet

numbers.
 Blood Groups
RBC membranes have glycoprotein antigens on their external
surfaces.

Karl Landsteiner, 1901 (Austrian scientist ) is the 1st to identify

blood groups of humans.
Found two antigens on the surface of RBCs and named them as

A antigen and B antigen.

 Are also called agglutinogens because of their capacity to cause

agglutination (clumping) of RBCs.

The corresponding antibodies (agglutinins) in the plasma and

named them anti-A or α- antibody and anti-B or β-antibody.

Importance of knowing blood group

 Medically, it is important during blood transfusions and in

tissue transplants.
 In pregnancy: helps to prevent the complications due to Rh

incompatibility and save the child from the disorders like

erythroblastosis fetalis.
 It has also legal importance in disputed paternity cases because

the erythrocyte antigens are inherited

 Blood group systems

The most known and clinically important antigens for blood grouping
are:
The ABO system of antigen and

The Rh system of antigen

 In the ABO system, blood is classified primarily on the basis of the

A and B antigens (also called agglutinogen) present on the surface

of RBC-membranes .
 People may have neither of them on their cells, they may have one,

or have both simultaneously

Cont,…

 Secondly, blood is classified on the basis of the naturally

occurring antibodies (agglutinins) in the serum.

 Based on the presence or absence of antigen A and antigen B,

blood is divided into four groups:

1. ‘A’ group
2. ‘B’ group
3. ‘AB’ group
4. ‘O’ group
Cont,…
 Blood having antigen A on the surface of RBCs is belongs to ‘A’

group. This blood has β- antibody in the serum.

 Blood with antigen B on the surface of RBCs is belongs to ‘B’ group.

This blood has α-antibody.

 If both the antigens are present on the surface of RBCs, blood group

is called ‘AB’ group and serum of this group does not contain any
antibody.
 If both antigens are absent on the surface of RBCs, the blood group is

called ‘O’ group and both α and β antibodies are present in the serum.
Cont,…
Determination of ABO Group
 Blood typing is done on the basis of agglutination.

 Agglutination occurs if an antigen is mixed with its

corresponding antibody which is called isoagglutinin.

Requisites for Blood Typing

i. A suspension of RBC which is prepared by mixing blood

drops with isotonic saline (0.9%)
Cont,…

i. Testing antisera are required

Antiserum A, containing anti-A or a-antibody.

Antiserum B, containing anti-B or ß-antibody.

 Presence of agglutination is confirmed by the presence of thick

masses (clumping) of RBCs

 Absence of agglutination is confirmed by clear mixture with

dispersed RBCs.
Cont,…
 Cont,…
Note: Positive reactions indicate agglutination
Universal donor
 Blood group “O” is called universal donor, because people

with this blood types have no antigens on their cell-membrane

surfaces and therefore cannot agglutinate if transfused to any
blood types.
 Even though they lack antigens, they have Anti- A & anti-B

antibodies in their plasma.

 So, they can receive blood only from persons with blood group

“O” .
Universal recipient
 Blood group AB is called universal recipient.

 People with AB-blood group can take blood from any blood

types, because they have no antibodies in their blood to cause

agglutination reactions.
 AB can donate blood only to a person with blood AB, not to

the others.
Blood transfusion
 Transfusion is widely used in clinical treatment.

 Transfusion of whole blood or its parts (plasma, or red blood

cell concentrates or other specific clotting factors) is important

to:
A. Restore blood volume

B. Improve O2 transport to tissues

C. Correct bleeding abnormalities etc.

Principle of Transfusion

 Principle of transfusion:

1. Identification of blood group must be taken before transfusion.

2. Cross-match test must be done before transfusion.
3.The same types of blood group for transfusion should be firstly
considered.
4. The different types of blood group for transfusion should be
very careful, small amount and slow import and if condition is
better, changes in the same types of blood group for transfusion.
Matching and cross-matching for blood transfusion

 Blood matching (typing) is a laboratory test done to determine

the blood group of a person.

 It is done to find out whether the person’s body will accept the

donor’s blood or not.

 For blood matching (typing)

Recipient’s RBC + Test sera

 Cross-matching is done by mixing the

Recipient’s serum + Donor’s RBC

Donators and Recipients

Donators
1. O can donate blood to group A, B, AB, and O
2. A “ A & AB only
3. B “ B & AB only
4. AB “ AB only

Recipients
1. O can receive blood from group O only
2. A “ A & O only
3. B B & O only
4. AB “ A, B, O, & AB
 Inheritance of ABO agglutinogens and agglutinins
Blood group of a person depends upon the two genes (A and B)
inherited from each parent.
 Are dominant by themselves and gene O is recessive.

Agglutinogens appear during the 6th month of fetal life.

Agglutinin αor β is not produced during fetal life.

 It appears 2–8 months after birth.

 It is produced in response to agglutinogens which enter the body

through respiratory system or digestive system along with bacteria.

Agglutinins are the gamma-globulins which are mainly IgG and IgM
immunoglobulins.
Cont,…
Inheritance of ABO group
Transfusion reactions due to ABO incompatibility (mismatched)

 Transfusion reactions occur due to transfusion error that involves

transfusion of incompatible (mismatched) blood.

 It occurs between donor’s RBC and recipient’s plasma.

 If the donor’s plasma contains agglutinins against recipient’s

RBC, agglutination does not occur.

 But, if recipient’s plasma contains agglutinins against donor’s

RBCs, the immune system launches a response against the

new blood cells.
 Donor RBCs are agglutinated resulting in transfusion reactions.
Rh Factor
 Rh factor is an antigen present in RBC.

 It was first discovered in Rhesus monkey and hence the name ‘Rh

factor’.
 There are many Rh antigens but only the D antigen is more

antigenic in human.
 The persons having D antigen are called ‘Rh positive’and those

without D antigen are called ‘Rh negative’.

 Rh group system is different from ABO group system because, the

antigen D does not have corresponding natural antibody (anti-D).

Cont,…
 These anti-Rehsus antibody was also found to a agglutinate

the majority (85%) of red cells of Humans.

 If Rh positive blood is transfused to a Rh negative person anti-

D is developed in that person.

 On the other hand, there is no risk of complications if the Rh

positive person receives Rh negative blood.

Transfusion Reactions due to Rh Incompatibility
 When a Rh negative person receives Rh positive blood for the

first time, he is not affected much, since the reactions do not

occur immediately.
 But, the Rh antibodies develop within one month.

 So, a delayed transfusion reaction occurs.

It is usually mild and does not affect the recipient.

But, when this person receives Rh positive blood for the second
time,
The donor RBCs are agglutinated and severe transfusion
reactions occur immediately.
Cont,…
Hemolytic Disease Of Fetus And Newborn –
Erythroblastosis fetalis

 Hemolytic disease is the disease in fetus and newborn,

characterized by abnormal hemolysis of RBCs.

Due to Rh incompatibility, i.e. the difference between the Rh

blood group of the mother and baby

 When a mother is Rh negative and fetus is Rh positive (the Rh

factor being inherited from the father), usually the first child
escapes the complications of Rh incompatibility.
because the Rh antigen cannot pass from fetal blood into the

mother’s blood through the placental barrier.

 Cont,…
 At the time of parturition (delivery), the Rh antigen from fetal blood

may leak into mother’s blood because of placental detachment.

 After delivery, the mother develops Rh antibody in her blood.

 When the mother conceives for the second time and if the fetus

happens to be Rh positive again, the Rh antibody from mother’s

blood crosses placental barrier and enters the fetal blood.
Causes agglutination of fetal RBCs resulting in hemolysis.
To compensate the hemolysis of more and more number of RBCs
is rapidly produced from bone marrow, spleen and liver
Many large and immature cells in proerythroblastic stage
are released into circulation
Cont,…
Leukocytes (White Blood Cells)
 Humans have direct contact to the external world.

 The Skin, respiratory system, GIT, eyes, urinary, genital organs etc.

 Infectious agents get access through them settle down in our body

 A lot of bacteria's harbor the body without causing diseases

 However, pathogenic microbes are defended by WBC’s through:

 A. Phagocytosis B. Production of sensitized lymphocytes &

 C. Antibody production
 (Cont,…) Leukocytes (White Blood Cells)
 Are the mobile units of the body’s immune defense system.

 Leukocytes, the only blood components that are complete cells

Compared to the RBCs, WBC s are :

 Are the colorless and nucleated formed elements of blood
 Larger in size and lesser in number (least numerous of blood cell)

 4000-10,000 / mm3 but, average no. is 7000 / mm3

 Leukocytes vary in structure, function, and number.

 Irregular in shape, many types and their lifespan is shorter

 largely use a “seek out and attack” strategy … go to sites of
invasion or tissue damage
 Classification WBC
 WBC are almost invisible under the microscope unless they are

stained.
 The stain used to identify white blood cells is usually mixture of a

pink-to-red stain ( eosin) and a blue-to-purple stain (basic stain).

 There are five different types of circulating leukocytes:
Neutrophils - Eosinophils,
Basophils -Monocytes, and Lymphocytes
Each with a characteristic structure and function.

 Some of the WBCs have granules in the cytoplasm others not,

when viewed microscopically.

Cont,…

 Depending on the appearance of their nuclei and presence or

absence of granules in their cytoplasm, the five types of

leukocytes fall into two main categories.
1. Granulocytes (polymorphonuclear granulocytes)
(neutrophils, eosinophils, basophils)
2. Agranulocytes (mononuclear agranulocytes)
(monocytes, and lymphocytes)
Genesis of WBC
 Originate from the same undifferentiated pluripotent stem cells that
also give rise to erythrocytes and platelets

A. Myelocytic line

- RBC, Platelets, Monocytes & Granulocytes

B. Lymphocytic lines

- T- lymphocytes and B-lymphocytes

 The granulocytes and monocytes are found only in the bone marrow.
 Lymphocytes (T and B cells ) are stored in lymphogenous tissues

(lymph glands, spleen, thymus, tonsils)

Cont,…
 Cont,…
 About two thirds of the circulating leukocytes are granulocytes,

mostly neutrophils.
 One third are agranulocytes, predominantly lymphocytes

 Depending on the type and extent of assault the body is combating,

leukocytes are selectively produced at varying rates.

 The rates of production of the various Leukocytes governed by:

 Chemical messengers arising from invaded tissue

 Bacterial or viral toxins

 Reaction products caused by plasma clotting in the inflamed area

 Life Spans Of Leukocytes
Lifespan of WBCs is not constant. It depends upon the demand in the body and their function.

Granulocytes and their life

a. Neutrophils (~ 62%), life span 4-5 days (are the most abundant WBC)
b. Eosinophils (~ 2-3%) “
c. Basophiles (~ 0.1-0.4%) “
Their life span become shortened to a few hours during serious infection.
Agranulocytes and their life span

i. Lymphocytes (~ 30%), circulate between the blood and lymph tissues

continually.
 Have life span of weeks or months depends of the body demand.
Cont,…

ii. Monocytes (~ 5%)

- Have short transit time to reach the tissue (10-20 hours)
- At this time, they are immature cells and have little ability
to fight infectious agents.
- Once they enter the tissue, they begin to swell and
increasing their diameters as much as five folds (60-80
µm) called macrophages (in this form they can live for
months).
 Capable of combating intratissue disease agents.
Cont,…
 Mechanisms Of WBC Mobility Through The
Tissues
1. Diapedesis: WBC Squeeze
through the pores of the blood
capillaries (Neutrophils and
Monocytes)
2. Amoeboid motion: Neutrophils
and Macrophages produce
pseudopodia and reach the
microbes in the tissues.
3. Chemotaxis: Is the attraction of
WBCs towards the injured tissues by
the (chemical, toxins, or degenerative
products of inflamed tissues)
4. Phagocytosis: Cellular ingestion
and destroying offending agent
(Neutrophils, macrophages)
How WBC differentiate normal cell from foreign invaders during
phagocitosis ?
 Smooth surfaces

Most natural structures in the tissues have smooth surfaces

which resist phagocytosis.

 Protective protein coats

Natural substances of the body have protective protein coats

that repel the phagocytes.

 Antibodies (immune system of the body)

Adhere to the bacterial membranes and thereby make the

bacteria susceptible to phagocytosis, (opsonization).

 Phagocytosis by neutrophils and
Macrophages
a. Presence of rough space on cells and attachment of antibodies
on bacterial surfaces (opsonization) facilitates phagocytosis

b. Dead tissues are easily phagotisized b/s they are not protected
by protein coat like the living cells

c. After phagocytosis, Lysosomes, and oxidizing agents like

lipases, peroxisomes, H2O2, etc. produced from cells kill
bacterial membranes.
Functions Of White Blood Cells
Neutrophils
 Are phagocytic specialists
(typically ingest and kill 5-20
bacteria)
 Their granules have enzymes
to digest bacteria
 Have 2-5 lobes and are the
most numerous of leukocyte
 Their numbers increase
during bacterial infection
 They stay in circulation for 10
hrs
 (Cont,… ) Eosinophils (cytotoxic cells)

 Nucleus is bilobed and are weak

phagocytes .
 Have larger granules in the
cytoplasm that have enzymes.
 Are found in the GI, lungs,
urinary and genital epithelial and
CT.
 Cannot engulf a much larger
parasitic worm, but they do
attach to the worm and secrete
substances that kill it. E.g.
Schistosoma
 Are assocoated with allergic rxn
and parasitic diseases.
 Their number increases during
asthma and other allergic attacks.
 Cont,…
Basophils
 Are rare in the circulation, constitute
< 1%
 Have coarse granules in the cytoplasm
and bilobed nucleus
 Are very similar with mast cell of
tissues.
 They destroy microorganisms by
releasing important substances from
their granules such as:
 heparin (inhibitors of blood
clotting)
 histamine, serotonin, cytokine
 Proteases and myeloperoxidase
 Cont,…
Monocytes
 Have no granules and are the largest
cells among the leukocytes.
 Are the precursors of the tissue
macrophages.
 Once enter the tissues from the blood
and they become tissue macrophages
-E.g. Kupffer cells in liver, alveolar
macrophages in lungs and
macrophages in spleen
 Play an important role in the
development of acquired immunity.
 Macrophages are highly phagocytic
cells that can ingest many bacteria
(up to 100), parasites and debris etc.
 Cont,…

Lymphocytes
 Have no granules and the nucleus

is not lobed
 Are responsible for Specific
immunity that consists of:

a. Cellular immunity (T
lymphocytes)

b. Humoral immunity (B
lymphocytes)
 Inflammation
Inflammation:

Is changes in body tissues that occurs after tissue injury .

 The injury can be caused by trauma, bacteria, heat etc.).

 The protective response of the tissues to the damage or destruction of

cells

Steps in inflammatory process

1. Damaged tissues

2. Chemicals like serotonin, histamine etc are released from damaged

tissue

3. Chemicals cause vasodilation, increased blood flow and enhanced

Capillary permeability
Cont,..

5. Fluids escape to interstitial space (fibrinogen causes clotting)

6. Migration of WBC (granulocytes and monocytes occur)

7.“Walling Off “ results.

It means separating the area of injury from normal tissues

Result:

a. If WBC’s win the battle, pus develops (dead pathogen+ WBC’s

+ tissue fluid)

b. If bacteria’s win and not walled off, then they enter into the
blood and cause septicemia
 Abnormalities In Leukocyte

Leukemia
 Increased numbers of abnormal white blood cells in the circulating blood.

 cancerous mutation of a myelogenous or lymphogenous cell

 Divided into two general types:

 Lymphocytic leukemias --cancerous production of lymphoid cells

 Myelogenous leukemias--cancerous production of myeloblast cells in

the bone marrow

 Their increased production takes the space of platelets & RBC causing

anemia + impaired blood clotting

Common effects of Leukemia
A. Metabolic starvation:
 The increased WBC consume too much metabolic substrates.

Thus energy source is depleted and too much use of amino

acids and vitamins by cancerous cells causes rapid
deterioration of other body tissues.

B. Development of infection, severe anemia and increased

bleeding tendency b/s of :
Displacement of RBC’s and thrombocytopenia (lack of

platelets) from bone marrows by the tumor cell.

Cont,…
Leucopenia
 Is decreased production of WBC
 Drug, such as chloramphenicol (an antibiotic), thiouracil
(used to
treat thyrotoxicosis), and various barbiturate hypnotics
 X-rays or gamma rays

 Symbiotic bacteria's that harbor the mucus can get access to our

body and cause toxicity, when marrow stops producing WBC’s.

- Differential count is usually undertaken in the lab to know the

proportions of WBC Numbers.
 Platelets
General Characteristics
 Are not whole cells but are small fragments that emerge from

megakaryocytes in red bone marrow (about 2-4 µm in diameter)

 Normal concentration of platelets in the blood: 1.5 x 105 – 3.5 x 105 /mm3

 Involved in blood clotting processes.

 Life Span 8-12 days

 Mostly have no nucleus

 Release serotonin, epinephrine and thromboxane (cause

vasoconstriction)
 Cont,…
 Thrombopoietin is the hormone that regulate platelet production.

 They have organelles and cytosolic enzyme systems for generating

energy and synthesizing secretory products (clotting protein, cytokines)

 They contain high concentrations of actin, myosin, thrombosthenin

which enable them to contract.

 The cell membrane of the platelets contain glycoproteins that repulses

adherence to normal endothelium

 They are always in inactive form the blood, when damage occur they

become active.
Cont,…
Hemostasis
 Is the arrest (cessation) of bleeding from a broken blood vessel

 Hemostasis involves four major steps:

(1) vascular spasm, (2) formation of a platelet plug (3) formation

blood cot
(4) formation of fibrous tissue into the blood clot to close the hole
 In vascular spasm small blood vessels and arterioles constrict at

local injured regions and decrease blood flow to injured areas.

 Platelet release its contents (serotonin, histamine, Thromboxane A 2)

at the injured place that initiate vasoconstriction.

Cont,..

Clotting process in a traumatized blood vessel.

 Cont,…

FIG.: States of hemostasis. ADP = Adenosine diphosphate; PAF = Plateletactivating

factor.
 Cont,…

Formation of a platelet plug

 Mechanism of Blood Coagulation
 Clotting takes place in
three essential steps:
1.Formation of prothrombin
activator
2.Conversion of
prothrombin into
thrombin
3. Convert fibrinogen into
fibrin fibers
Cont,…

Generally activator substances for clotting are from:

a. The injured vascular walls

b. Platelets &

c. Blood plasma near the injured site initiate cascade of

reactions to clot the wound.
 Most of the chemicals involved in clot formation are

synthesized in the liver.

 Cont,…
Clotting Factors: Formation of Fibrin
 Clots is composed of a meshwork of fibrin fibers contain platelets,
blood cells and plasma
 The conversion of fibrinogen into fibrin occur via either of two

pathways by initiating prothrombin activator

 Intrinsic pathway - produces clots when collagen is exposed to

plasma.
Extrinsic pathway - produces clots when damaged vascular wall

and surrounding tissues release a chemicals (tissue

thromboplastin or factor III) that initiate formation of fibrin
Cont,… Clotting factors
 Cont,…
Intrinsic Pathway
 Begins with the
exposure of the blood to
collagen from a
traumatized blood vessel
wall
 Involves five separate
steps
 Cont,…
Extrinsic Pathway
 Begins when a
traumatized
vascular wall or
extravascular
tissues that come in
contact with the
blood
Cont,..

 Formation of fibrin also require the presence of Ca 2+and

phospholipids
convert inactive glycoprotein, called prothrombin, into the

active enzyme thrombin.

Thrombin converts the soluble protein fibrinogen into fibrin

monomers.
Cont,…
 Dissolution of Clots
 As the damaged blood
vessel wall is repaired
 The clot is no longer needed
to prevent hemorrhage
 So, it is slowly dissolved by
a fibrinolytic (fibrin-
splitting) enzyme called
plasmin.
 The dissolution of fibrin by
plasmin known as
fibrinolysis.
 Phagocytic white blood cells
gradually remove the
products of clot dissolution.
 Bleeding disorders

Excessive bleeding can be caused due to 3-major factors:

1. Hemophilia: occurs almost exclusively in males.

- Due to insufficiency of Factor VIII and IX

2. Vit. K deficiency: (liver diseases, hepatitis, cirrhosis)

Vit. K is important to synthesize Prothrombin in the liver, VII,

IX, X and protein C

3. Thrombocytopenia (platelet deficiency)