Blood and

Body Fluid

Dr Ogedengbe J. O.
Department of Human Physiology,
College of Health Sciences,
University of Abuja
 Introduction

◼ Body is made up of solids and fluids.

◼ Fluid part is more than two third of the

whole body.
◼ Water forms most of the fluid part of the
body.
◼ In human beings, the total body water
varies from 45% to 75% of body weight.
Percentage of H2o in Tissues
 Blood = connective tissue

extracellular specialized cells:

cells
matrix: (Formed elements)
Plasma RBCs
WBCs
Platelets
 FLUID COMPARTMENTS

EXTRA CELLUAR INTRA CELLULAR

FLUID - 18L (cytosol) FLUID – 22L

PLASMA INTERSTITIAL TRANSCELLULAR

2.75L FLUID – 12L FLUID – 3.25L

1. CSF
2. Intra ocular
3. Pleural
4. Peritoneal
5. Synovial
6. Digestive Secretions
Serum = Plasma –
Clotting factors
Body Fluid Compartments

Body fluid compartments

and movement of fluid
between different
compartments. Other fluids
= Transcellular
fluid, fluid in bones and
fluid in connective tissue.
 Intracellular
Plasma Interstitial Fluid
Fluid

Organic anion
 Significance of Body
Fluids
◼ In Homeostasis
◼ In Transport Mechanism
◼ In Metabolic Reactions
◼ In Texture of Tissues
◼ In Temperature Regulation
 Determination of Body
Fluids
◼ INDICATOR DILUTION METHOD
◼ Principle
◼ Formula to Measure the Volume of Fluid
V = M/C
V= Volume of fluid in the compartment.
M= Mass or total quantity of marker substance
injected.
C = Concentration of the marker substance in
the sample fluid
Corrected: Volume = M – Amt. of subst. excreted/C
 DILUTION PRINCIPLE
Principle of mass conservation
Based on using a marker whose concentration can be
measured.

Inject x gm of marker into compartment

measure concentration at equilibrium (y gm/L)
Since concentration = mass/ volume
Volume = mass / concentration
= x/y L

C1V1=C2V2
 Characteristics of Marker
Substances
1. Must mix evenly throughout the compartment
2. Non toxic, no physiological activity
3. Even mixing
4. Must have no effect of its own on the distribution
of water or other substances in the body
5. Either it must be unchanged during the
experiment or if it changes, the amount changed
must be known.
6. The material should be relatively easy to
measure.
 Indicators Used For Measuring Plasma
Volume, ECF Volume And Total Body H2O
Compartmen Criterion Indicators
t
1. Plasma Substance 1. Evans blue dye;
should not cross 2. radioiodinated
capillaries fibrinogen;
3. radioiodinated
albumin
2. ECF Substance 1. Radioactive Na, Cl, Br,
volume should cross sulfate & thiosulfate.
capillaries but 2. Non-metabolizable
not cross cell saccharides like inulin,
membranes mannitol, & sucrose
3. Total body Substance Heavy H O, tritiated H O,
 Measurement of Total
Body Water
◼ Deuterium oxide, tritium oxide and antipyrine
mixes with fluids of all the compartments
within few hours of injection.

◼ Since plasma is part of total body fluid, the

concentration of marker substances can be
obtained from sample of plasma.
 Measurement of
Extracellular Fluid Volume
◼ Uses substances that pass through the
capillary membrane but do not enter the cells.
◼ The substances remain only in ECF and do
not enter the cell.
◼ Since ECF includes plasma, the concentration
of marker substance can be obtained in the
sample of plasma.
 Compartments with no
Compartment-Specific Subst.
Determine by subtraction:
◼ How would you measure ICF volume?
◼ Cannot be measured; it is calculated
(estimated).
ICF volume = Total body H2O – ECF volume

◼ Interstitial volume
◼ Can not be measured directly
Interstitial Fluid Volume (ISFV).
ISFV = ECFV - PV
 Concentration of Body
Fluids
◼ Concentration of body fluids is expressed
in three ways:

1. Osmolality
2. Osmolarity
3. Tonicity
 Osmolality (Osmotic Conc)

◼ Measureof a fluid’s capability to

create osmotic pressure.
Osmolality is expressed as the
number of particles (osmoles)
per kilogram of solution
(osmoles/kg H2O).
 Osmolarity

◼ Osmolarity is another term to express the

osmotic concentration. It is the number
of particles (osmoles) per litre of solution
(osmoles/L).
 Tonicity

◼ It is the measure of effective osmolality.

In terms of tonicity, the solutions are
classified into three categories:
i. Isotonic fluid
ii. Hypertonic fluid
iii. Hypotonic fluid.
Any Question?
 Properties of Blood
1. Color
2. Volume: Average volume of blood in a normal adult
is about 5 L
3. Reaction and pH: Blood is slightly alkaline and its
pH in normal conditions is 7.4
4. Specific gravity:
➢ Specific gravity of plasma : 1.022 to 1.026
➢ Specific gravity of total blood : 1.052 to 1.061
➢ Specific gravity blood cells : 1.092 to 1.101
5. Viscosity: Blood is five times more viscous than
water.
 Variations In Blood Volume

◼ Physiological Variations:
1. Age
2. Sex
3. Body Weight
4. Atmospheric Temperature
5. Pregnancy
6. Exercise
8. High Altitude
9. Surface Area of the Body
10. Emotion
11. Sleep
12. After meals
13. High barometric pressures
◼ Pathological Variations:
Hypervolemia-
1. Hyperthyroidism
2. Hyperaldosteronism
3. Cirrhosis of the Liver
4. Congestive Cardiac Failure
Hypovolemia-
1. Haemorrhage or blood loss
2. Fluid loss
3. Haemolysis
4. Anaemia
5. Hypothyroidism
 Functions of blood
1. Breathing function of blood.
2. Trophic function of blood.
3. Excretory function of blood.
4. Hormonal regulation.
5. Temperature regulation.
6. Maintaining the acid-base balance of tissue.
7. Supporting the water-electrolytic balance.
8. Homeostasis function.
9. Protecting the body from bacteria and other
organisms that can cause diseases or other
abnormal conditions.
 Plasma

◼ Liquid part of blood

◼ Contains:
◼ Water
◼ Proteins
◼ Hormones
◼ Nutrients
◼ Electrolytes
◼ Respiratory Gases
◼ Wastes
 Plasma Proteins

◼ Include:
◼ Albumin (60%)
◼ Globulins (36%)
(Antibodies &
transport proteins) Plasma
◼ Clotting Proteins
(4%)
◼ Enzymes &
Hormones
 Serum
Liquid part of blood without clotting proteins

◼ Albumin (60%)
◼ Globulins (36%)
(Antibodies &
transport proteins)
X
◼ Clotting Proteins
(4%)
Plasma
Serum

Clot
◼ Enzymes &
Hormones
 Properties of Plasma
Proteins
◼ Molecular Weight
◼ Albumin: 69,000
◼ Globulin: 156,000
◼ Fibrinogen: 400,000

◼ Oncotic Pressure
◼ colloidal
oncotic pressure is about
25mm Hg. Albumin plays a major role.
◼ Specific Gravity
 Albumin

◼ Itis the most abundant plasma

protein - it provides oncotic
pressure, It connects and
transports bilirubin, fat acids,
antibiotics and sulfanilamids. It
is produced in liver in average
quantity of 17 gram per day.
 Globulin

◼ Produced in lymphatic nodes, in liver and

in bone marrow in average quantity of 5
gram per day.
◼ α-1-globulins connects with
carbohydrates to form glyco-proteins.
◼ α-2-globulins connects 90 % of copper.
This forms cerruloplasmin.
◼ β-globulin carries fats, iron (for example,
transferrin).
◼ ɣ-globulins has protective functions (for
example, antibodies).
 Fibrinogen
◼ Fibrinogen is protein which are
produced by liver and take part in
hemostasis.
◼ Fibrinogen is soluble which becomes
insoluble when transformed to fibrin
and provide coagulative hemostasis
(plug production) and prevent
bleeding.
◼ Daily prod. of fibrinogen is 2-4 g/L.
Any Question?
 Red Blood Cells

◼ Red blood cells are the non-nucleated

formed elements in the blood
◼ Colour of the RBC is due to the
colouring pigment called hemoglobin.
◼ They are the most populous of the
three cellular components of the blood.
◼ Normal value ranges between 4 and 5.5
million mm3 of blood.
 Red Blood Cells

◼ Shape & size

◼ Flat Biconcave Disc
◼ Non-nucleated
◼ Diameter 7-8microM x 2.5mm x
1mm
◼ Average volume 90-95 mm3
◼ Flexible
◼ Number = 4.7-5 x106
◼ Hb = 34g/dl of cells
◼ Hb = 14-16 g/dl in the whole
blood
◼ Men: 13.8 to 18.2 g/dl
◼ Women: 12.1 to 15.1 g/dl
◼ Children: 11 to 16 g/dl
◼ Pregnant women: 11 to 12 g/dl
 Production of RBC

◼ In the first few weeks of embryonic life,

nucleated RBCs are formed in yolk sac.
◼ By middle trimester, mainly in liver,
spleen & lymph nodes.
◼ In the 3rd trimester, RBCs are formed in
bone marrow of all bones
◼ Bone marrow of flat bone continue to
produce RBC into adult life
◼ Shaft of long bone stop to produce RBC
at puberty while epiphysis continued
Production of RBC
 Genesis of RBC
◼ All blood cell are formed from
Pluripotential hematopoietic stem cells 
committed cells:

◼ Committed stem cells for RBC

◼ Committed stem cells for WBC

◼ Growth of different stems cells are

controlled by different growth factors
 Stages of differentiation of
RBC
◼ Stages of RBC development
◼ Committed stem cell
◼ Proerthroblast
◼ basophil erythroblast
◼ polychromatophil erythroblast
◼ orthochromatic erythroblast
◼ Reticulocytes
◼ Mature erythrocytes
◼ Rapid RBC production →  reticlocytes in
the circulation
 Erythropoiesis

◼ RBC development is
characterize by:

◼ decrease in cell size

◼ disappearance of nucleus
◼ appearance of haemoglobin
 Regulation of RBC production
◼ Erythropoiesis is stimulated by
erythropoietin hormone produced by the
kidney in response to hypoxia (low
oxygen in the blood)
◼ Hypoxia caused by:
◼ Low RBC count (Anaemia)
◼ Haemorrhage
◼ High altitude
◼ Prolong heart failure
◼ Lung disease
Tissue oxygenation and
RBC formation
 Erythropoietin
◼ Glycoprotein
◼ 90% from renal cortex and 10% from liver
◼ Stimulate the growth of early stem cells
◼ Does not affect maturation process
◼ Can be measured in plasma & urine
◼ High levels of erythropoietin
◼ anemia
◼ High altitude
◼ Heart failure
◼ Lung Disease

(Result in polycythemia)
Role of the kidneys in RBC
formation
 Formation of Haemoglobin

◼ Synthesis begins in the proerythroblasts

and continues into the reticulocyte stage
of the red blood cells.

◼ Thus, when reticulocytes leave the bone

marrow, they continue to form minute
quantities of Hb until they become
mature erythrocytes.
◼ Succinyl-CoA, binds with glycine to form a
pyrrole molecule.
◼ In turn, four pyrroles combine to form
protoporphyrin IX, which then combines
with iron to form the heme molecule.
◼ Finally, each heme molecule combines with
a globin synthesized by ribosomes, forming
a subunit of Hb.
◼ Four of these in turn bind together loosely
to form the whole Hb molecule.
◼ There are different types of chains
designated α-chains, β-chains, ɣ-
chains, and δ-chains.
◼ The most common form of Hb in the
adult human being, Hb A, is a
combination of two α-chains and two
β-chains.
◼ Hb A has a molecular weight of 64,458.
◼ There are four iron atoms in each Hb
molecule; each of these can bind
loosely with one molecule of O2,
making a total of four molecules of
O2 that can be transported by each
Hb molecule.
◼ The types of Hb chains determines
the binding affinity of the Hb for O2
◼ Abnormalities of the chains can alter the
physical characteristics of the Hb molecule
◼ E.g., in sickle cell anaemia, the amino acid
valine is substituted for glutamic acid at one
point in each of the two β-chains.
◼ This alters the morphology of the cell and
easy to lyse.
◼ The most important feature of the Hb
molecule is its ability to combine loosely and
reversibly with O2
 IRON INTAKE & METABOLISM
◼ The total of iron in the body is 4 to 5 grams
◼ About 65% of which is in the form of
hemoglobin.
◼ About 4% is in the form of myoglobin.
◼ 0.1% is combined with the protein
transferrin in the blood plasma,
◼ 15-30% is stored, mainly in reticulo-
endothelial system and liver cells,
principally in the form of ferritin.
 Transport and Storage of
Iron
◼ From the small intestine, it moves out to
combines a β-globulin, apotransferrin, to
form transferrin, which is then transported
in the plasma.
◼ The iron is loosely bound in the transferrin
&, can be released at any point in the body.
◼ Excess iron in blood is deposited in the
liver cell and reticuloendothelial cells of the
bone marrow.
◼ In the cell cytoplasm, iron combines
apoferritin, to form ferritin.
◼ This iron stored as ferritin is called storage
iron
◼ Smaller quantities of the iron in the storage
pool are in an insoluble form called
hemosiderin
◼ This is occurs when the total quantity of
iron in the body is more than the apoferritin
storage pool can accommodate
◼ Transferrin binds strongly with
receptors in the cell membranes of
erythroblasts in the bone marrow.
◼ Then, it is ingested into the
erythroblasts by endocytosis.
◼ It is delivered directly to the
mitochondria, where heme is
synthesized
???
 Anemia
Reduced O2 carrying capacity of the blood
◼ Insufficient number of RBCs:
Hemorrhagic - due to blood loss associated with an
injury, undiagnosed bleeding ulcer, etc.

Hemolytic - due to blood loss due to transfusion

reactions & certain bacterial and parasite infections.

Aplastic - due to destruction or inhibition of red

marrow by drugs, ionizing radiation or certain bacterial
toxins.
 Anemia

◼ Insufficient hemoglobin content in

RBCs:
Iron Deficiency - inadequate intake or absorption of
iron.

Pernicious - dietary deficiency of Vitamin B12 or inadequate

production of intrinsic factor for absorption of Vitamin B12.
 Anemia

◼ Abnormal hemoglobin in RBCs:

Sickle Cell - one amino acid in the 287 forming the beta
chains is wrong.

In low O2 conditions the beta chains form stiff rods

which cause RBCs to sickle blocking small vessels.
 Other Blood Related
Disturbances
◼ Polycythemia - abnormally high
number of RBCs (8 - 11 million/mm3).
Increases blood viscosity & blood
pressure.
◼ Cause - most often the result of bone
marrow cancer.
• Lecuopenia - abnormally low number
of WBCs (less than 5,000/mm3).
• Cause - drugs, steroids & anti-cancer
agents.
 Other Blood Related
Disturbances
◼ Leukemia - abnormally high numbers
of immature WBCs that are mitotic &
unspecialized.
◼ Named according to abnormal WBC
type involved
◼ Myelocytic - derived from myeloblasts
(chronic)
◼ Lymphocytic - involves lymphocytes
(acute)
 Other Blood Related
Disturbances
◼ Thrombus - a clot that forms & is stationary
in an unbroken blood vessel.
◼ If sufficiently large, it may block the flow of
blood downstream causing death of those
tissues.

• Embolus - an abnormal object moving through a

blood vessel, ie. Clot, air bubble, lipid droplet, etc.
• Embolism - a blockage of blood vessels caused by an
embolus. May cause a stroke or heart attack
depending on tissues affected.
 Haemostasis & Blood
Coagulation
◼ Haemostasis means stoppage of bleeding and
prevention of blood loss when a vessel is
injured.
◼ There are 4 mechanism invloved
1. Vasoconstriction of injured vessel: occurs due
to
I. Nervous reflex
II. Myogenic contraction of the vessel
III. Release of serotonin & thromboxane A2 from platelets
(both cause Vasoconstriction)
2. Formation of platelet plug(temporary
haemostatic plug):
Platelets stick to parts of damaged blood
vessel, become activated and accumulate
large numbers
3. Blood clotting (definitive haemostatic
plug)
4. Repair of damaged blood vessel
 Red blood cell

Platelet

Collagen fibers
and damaged
endothelium

Platelet Plug 11 Platelet adhesion

Formation

Liberated ADP,
serotonin, and
thromboxane A2

22 Platelet release reaction

Platelet plug

33 Platelet aggregation
 Platelets (Thrombocytes)

◼ They are small granulated round bodies

2-4 microns in diameter
◼ They have no nuclei
◼ Normal count is 200,000-500,000/mm3
◼ Life span averages 8 days
◼ Normally 60-75% are present in
circulating blood, remainder are present
in the spleen.
 Functions of the Platelets

1. Formation of the primary haemostatic plug

2. Release of vasoconstrictor subs, serotonin
& thromboxane A2
3. Release of platelet factor 3 which is
essential for blood clotting
4. Stabilization of the blood clot (factor XIII)
and induction of clot retraction
5. Release of the PDGF which repairs the
damaged vessel wall.
Blood clotting:
Serum is blood plasma minus clotting
proteins
 Clotting

Series of
chemical
reactions
culminating
in formation
of fibrin
threads
 Clotting
(coagulation)
factors – Ca2+ ,
several inactive
enzymes, various
molecules
associated with
platelets or released
by damaged tissues
 Factors Required for Blood
Clotting
◼ Factor I – Fibrinogen
◼ Factor II - Prothrombin
◼ Factor III – Tissue thromboplastin, Tissue
factor
◼ Factor IV – Calcium Ion
◼ Factor V – Proaccelerin, labile factor
◼ Factor VII – Proconvertin, stable factor
◼ Factor VIII – Antihaemolytic globulin or
factor
◼ Factor IX – Christmas factor
◼ Factor X – Stuart Prower factor
◼ Factor XI – Plasma thromboplasmin
antecedent
◼ Factor XII – Hageman factor (glass or
contact factor)
◼ Factor XIII – Fibrin-stabilizing factor
◼ High molecular weight kininogen –
Fitzgerald factor
◼ Prekallikrein – Fletcher factor
◼ Platelet factor 3 – Platelet thromboplastin
(phospholipids)
◼ von Willebrand factor – Factor VIII-related
antigen
 The Cascade Mechanism
(Theory) of Blood Clotting
◼3 Stages of Clotting
◼ Extrinsic or intrinsic pathways lead
to formation of prothrombinase
◼ Prothrombinase converts
prothrombin into thrombin
◼ Thrombin converts fibrinogen
(soluble) into fibrin (insoluble)
forming the threads of the clot
(a) Extrinsic pathway (b) Intrinsic pathway

Tissue trauma Blood trauma

Damaged
endothelial cells
expose collagen
fibers

Tissue
factor
(TF)

Damaged
platelets

Activated XII

Activated
Ca2+ Ca2+
platelets
+
Platelet
phospholipids

Activated X Activated X

V V +
Ca2+ Ca2+
1 PROTHROMBINASE
(c) Common
pathway

Ca2+
Prothrombin
(II)
THROMBIN 2
Ca2+ XIII

Fibrinogen
(I) Activated XIII
Loose fibrin STRENGTHENED 3
threads FIBRIN THREADS
 Blood Clotting
◼ Extrinsic pathway
◼ Fewer steps than intrinsic and
occurs rapidly in 15-20sec
◼ Tissue factor (TF) or
thromboplastin leaks into the
blood from cells outside (extrinsic
to) blood vessels and initiates
formation of prothrombinase
◼ Intrinsic pathway
◼ More complex and slower than
extrinsic (begins in 1-2min and
terminates in 3-6mins)
◼ Activators are either in direct contact
with blood or contained within (intrinsic
to) the blood
◼ Outside tissue damage not needed
◼ Also forms prothrombinase
 Blood Clotting: Common
pathway
◼ Marked by formation of
prothrombinase
◼ Prothrombinase with Ca2+
catalyzes conversion of
prothrombin to thrombin
◼ Thrombin with Ca2+ converts
soluble fibrinogen into insoluble
fibrin
◼ Thrombin has 2 positive feedback
effects
◼Accelerates formation of
prothrombinase
◼Thrombin activates platelets
◼Clot formation remains localized
because fibrin absorbs thrombin and
clotting factor concentrations are low
 Role of Calcium in Blood
Clotting
◼ It’s an essential catalyst in bld. clotting
◼ The normal serum level is 2.25-2.5
mmol/L
◼ If removed from bld. clotting will not
occur
◼ However in vivo reduction of blood
calcium to level that stops blood
clotting is incompatible with life.
 Importance of Vitamin K in
Blood Clotting
◼ Vitamin K is a fat-soluble vitamin
◼ It is a cofactor for the enzyme that
catalyzes conversion of glutamic acid to
gamma-carboxyglutamic acid.
◼ Six of the proteins involved in clotting
requires this conversion.
◼ These include factors II, VII, IX and X as
well as proteins C and S.
◼ Thus it is called antihaemorrhagic vitamin
 Causes of Vitamin K
Deficiency
◼ Failure of absorption – e.g. due
to deficient bile flow (obstructive
jaundice)
◼ Deficient intestinal bacteria
(prolonged use of antibiotics and
in newly born infant)
◼ Severe lack of the vitamin in diet
 Factors that Affect Blood
Clotting
◼ Factors that promote clotting
1. In vitro (a) warming of blood (b)
contact with negatively charged or
wettable surfaces eg glass (c) addition
of foreign body into blood
2. In vivo (a) blood stagnation (b)
roughening or damage of epithelial
lining (c) injection of vit K or
adrenaline
◼ Factors that inhibit clotting
1. In vitro
1. (a) cooling of blood
2. (b) collection of blood in non-wettable
vessels eg paraffin or silicone-coated test
tubes
3. (c) blood defibrination
4. (d) addition of heparin
5. (e) decreasing the blood Ca2+
2. In vivo (a) vit K deficiency (b)
small dose of asprin (c)
administration of heparin or
dicumarol
 Anticoagulants

A. Anticoagulants used in vitro

1. Sodium oxalate
2. Chelating agents eg Na citrate and EDTA
(ethyline-diamine tetra-acetic acid)
3. Heparin (used both in vivo & in vitro)
B. Anticoagulants used in vivo
1. Heparin
2. Dicumarol
 Natural Anticlotting
Mechanisms
◼ Normally, blood clotting is prevented (so
that fluidity is maintained) by the
following mechs
1. Rapid blood flow rate and the smooth &
intact endothelium
2. The balance btw the effects of
thromboxane A2 and PGI2
3. Synthesis of various clotting factors in
inactive forms
4. The low conc of various clotting factors,
& their rapid removal by the RES in the
liver
5. Presence of heparin & antithrombin III
6. Adsorption of any thrombin formed to the
fibrin threads (which inhibits its effect,
thus preventing excessive clotting)
7. The fibrinolytic system that continuously
removes small clots
Fibrinolytic System &
Regulation by Protein C
Thrombomodulin Endothelial Cell
Thrombin
Protein C Activated Prot C (APC)

VIIIa Inactive VIII Va Inactive V

Inactivates inhibitor of tissue plasminogen activator

Plasminogen Plasmin
Lyses fibrin
 Blood Clotting Process
◼ Platelets - Form temporary plug &
release platelet factors which
catalyze clot formation
 ◼ Platelet factors - React with
Calcium (Ca2+) & other clotting
factors in the plasma to initiate
clot formation.

• Thromboplastin - a lipid (Tissue Factor) released from injured cell

membranes which accelerates the clotting process.
• Lipids released from damaged cell membranes such as
thromboplastin having a localized effect are called Prostaglandins.
◼ Platelet factors, Ca2+ & other clotting factors
in plasma initiate clot formation
• A plasma protein (Prothrombin) is converted by
prothrombin activator into an enzyme Thrombin.
◼ Thrombin converts the plasma protein
Fibrinogen into the insoluble protein
Fibrin.
◼ Fibrin forms a mesh which glues the
platelets & RBCs together to form the clot.
 250,000-
500,000/mm3

4 - 6 million/mm3
 White Blood Cells/
Leukocytes
◼ Have nuclei
◼ Do not contain hemoglobin
◼ Granular or agranular based on staining
highlighting large conspicuous granules
◼ Granular leukocytes
◼ Neutrophils, eosinophils, basophils
◼ Agranular leukocytes
◼ Lymphocytes and monocytes
Types of White Blood Cells
 Functions of WBCs
◼ Usually live a few days
◼ Except for lymphocytes – live for months or
years
◼ Far less numerous than RBCs
◼ Leukocytosis is a normal protective response to
invaders, strenuous exercise, anesthesia and
surgery
◼ Leukopenia is never beneficial
◼ General function to combat invaders by
phagocytosis or immune responses
 Emigration of WBCs
◼ Many WBCs leave the
bloodstream
◼ Emigration (formerly
diapedesis)
◼ Roll along endothelium
◼ Stick to and then
squeeze between
endothelial cells
◼ Precise signals vary for
different types of WBCs
 Granulocyte - Neutrophil
◼ Nucleus with 3 to 6 lobes
• Cytoplasmic granules fine, both basic & acidic
• Phagocytize bacteria &
some fungi
• Attracted by chemotaxis
• Respond most quickly to
tissue damage by bacteria
• Produced in bone marrow
by myeloblasts
Phagocytosis
 Granulocyte - Eosinophil
◼ Nucleus bi-lobed
◼ Cytoplasmic granules coarse &
acidic (red/orange)

Leaves capillaries
and enter tissue fluid

• Destroy parasitic worms & immune complexes

• Produced in bone marrow by myeloblasts
 Granulocyte - Basophil
◼ Nucleus lobed - U or S shaped
◼ Cytoplasmic granules large & basic
-(purplish/black)

❑ Intensify inflammatory
reaction
❑ Involved in
hypersensitivity
reactions (allergies)

• Cause vasodilation by the release of histamines

• Produced in bone marrow by myeloblasts
 Agranulocyte - Lymphocyte
◼ Nucleus spherical-fills half or more
of cell
• No visible granules in cytoplasm
• The major soldiers of
the immune system
• B lymphocytes -
Humoral Immunity
(antibodies)
• T lymphocytes - Cellular Immunity
• Natural Killer (NK) cells - microbes and certain
tumor cells
• Produced in lymphatic tissues
 Agranulocyte - Monocyte
◼ Nucleus U or kidney shaped -fills
half or more of cell
• No visible cytoplasmic granules
• Takes longer to arrive but
arrive in larger numbers and
destroy more microbes
• Differentiate into macrophages in tissues.
• Provide defense against viruses & intracellular
bacteria in chronic infections.
• Activate lymphocytes
• Produced by monoblasts in lymphatic tissues.
 Two Types of Immunity

◼ Innate (Natural or Non-specific) immunity

◼ “possessed at birth, possessed as an essential
characteristic”
◼ Always present

◼ Adaptive (Specific) immunity

◼ “to make suitable to or fit to a specific use or
situation”
◼ Created and modified
 Innate Immunity

◼ Protection by Skin and Mucous

Membranes
◼ Phagocytic Cells
◼ Remove debris (garbage men)
◼ Macrophages, Neutrophils, Monocytes
◼ Natural Killer Cells
◼ Lymphocytes that kill virally infected cells and
tumours
◼ Complement System
◼ “complements antibody in the killing of bacteria”
◼ A group of >30 proteins found in the blood
 ACQUIRED IMMUNITY OR
SPECIFIC IMMUNITY

◼ Acquired immunity is the resistance

developed in the body against any
specific foreign body like bacteria,
viruses, toxins, vaccines or
transplanted tissues
Types of White Blood Cells
◼ There are 5 different types of WBCs
◼ Neutrophils (60%)
◼ kill bacteria
◼ Eosinophils (2%)
◼ Allergic response
◼ Parasite killing
◼ Basophils (1%)
– Allergic reactions
◼ Monocytes (4%)
◼ Become macrophages
◼ Lymphocytes (33%)
◼ Direct the immune system
 Lymphocytes

◼ Two types of lymphocytes

◼ T-Cells (Thymus derived)
◼CD4+ T-Cells (helper, inducer T cells)
◼CD8+ T-Cells (cytotoxic, killer T cells)
◼Suppressor T-Cells
◼Memory T-Cells

◼ B-Cells (Bone Marrow derived)

 Types of B Lymphocytes
After processing, the B lymphocytes are
transformed into two types:
◼ 1. Plasma cells
◼ They destroy foreign organisms by producing
the antibodies
◼ 2. Memory cells
◼They are inactive until the body is exposed to
the same organism for the second time.
◼ This phenomenon forms the basic principle of
vaccination against the infections
 Adaptive Immunity
◼ Two Components of Adaptive
Immune System
◼ Humoral (humoral mediated
immunity)
◼ B-Cells ➔ Plasma Cells ➔ Antibodies
◼ Cellular (cellular mediated immunity)
◼ CD8+ T-Cells ➔ Direct Cellular Killing
◼ CD4+ T-Cells ➔ Recruitment of other
immune cells (inflammatory response)
 Immune Response-Antigen

◼ Antigen – “any substance when

introduced into the body stimulates
the production of an antibody”
◼ Bacteria, fungus, parasite
◼ Viral particles
◼ Other foreign material
◼ Pathogen – an Antigen which
causes disease
 Immune Response -
Antibodies
◼ Antibody – “a Y-shaped protein, found
on the surface of B-Cells or free in the
blood, that neutralize antigen by binding
specifically to it”
◼ Also known as an Immunoglobulin

Antigen
Humoral Mediated
Immunity
 Cellular Mediated Immunity

◼ ViaT-Cells
◼ CD8+ T-Cell
◼ Stimulated ➔ Direct Killing
◼ CD4+ T-Cell
◼ Th1 ➔ Stimulated ➔ Macrophage
Activation
◼ Th2 ➔ Stimulated ➔ B-Cell Activation
 Cellular Mediated Immunity

◼ Remember B-Cells have direct surface

receptors (immunoglobulins) for antigen!
◼ T-Cells do not possess these receptors
◼ Instead, T-Cells need to have antigen
presented to them (like on a silver platter)
◼ Antigen is presented to T-Cells by …
Antigen Presenting Cells
 Cellular Mediated Immunity

◼ TwoGeneral Professional
Types of Antigen Presenting Cells
APC
(APCs) APC
All Cells B-Cells, Macrophages,
Dendritic Cells
Present antigen found inside Present antigen found
the cell outside the cell
Use an MHC class I Use an MHC class II
molecule to present antigen molecule to present antigen
Interact with CD8+ T-Cells Interact with CD4+ T-Cells
➔ Cellular Killing ➔T-Cell Help
 General APCs

◼ All cells in the body are always

“cleaning” themselves
◼ When they find some “dirt” (viral
protein, normal cellular debris) ➔ Need
to make sure it is not something
harmful
◼ Attach the “dirt” to an MHC-I molecule
◼ Present this “dirt” to a CD8+ T-Cell
General APCs &
CD8+ T-Cells
 Professional APCs

◼ Professional APCs have the ability to

take up (endocytosis) extracellular
proteins (self or foreign)
◼ Break down this protein into peptides
and attach it to an MHC-II molecule
◼ Present the peptide to a CD4+ T-Cell
Professional APCs
CD4+ Th1-Cells
Professional APC
CD4+ Th2-Cells
 Summary of Adaptive
Immunity
◼ Humoral
◼ Antibody Production – B-Cells
◼ Cellular
◼ CD8+ T-Cells ➔ MHC-I ➔ Cytotoxic
◼ CD4+ Th1-Cells ➔ MHC-II ➔ Activate
Macrophages
◼ CD4+ Th2-Cells ➔ MHC-II ➔ Activate B-Cells
to produce Antibody
 What Prevents the Body
from Attacking Itself?
◼ Two Concepts
◼ Central Tolerance
◼ Peripheral Tolerance
 Central Tolerance

◼ Occurs during lymphocyte (T & B Cells)

maturation in the primary lymphoid organs
(thymus & bone marrow)
◼ The body presents immature lymphocytes with
self-antigen
◼ Lymphocytes which react with high affinity to
this self-antigen are deleted (apoptosis)
◼ Lymphocytes which react with low affinity are
positively selected to mature
Central Tolerance
 Peripheral Tolerance
◼ During maturation, lymphocytes cannot be
presented with every self-antigen
◼ Some antigens are found in low concentrations
in specific locations
◼ New antigens are formed during life
◼ Therefore, lymphocytes come in contact with
new antigen
◼ Particular importance to the cytokine
environment present when lymphocytes
encounter this new antigen
 MEMORY T CELLS
◼ Some of the T cells activated do not enter
the circulation but remain in lymphoid
tissue.
◼ These are called memory T cells.
◼ Later they migrate to various lymphoid
tissues throughout the body.
◼ When the body is exposed to the same
organism the 2nd time, these cells identify
them and immediately activate the other T
cells.
 Types of Antibodies

◼ Five types of antibodies are identified:

1. IgA (Ig alpha)
2. IgD (Ig delta)
3. IgE (Ig epsilon)
4. IgG (Ig gamma) 75% of the antibodies
5. IgM (Ig mu).
 Functions of Different
Antibodies
◼ 1. IgA - localized defense mechanism
in external secretions like tear
◼ 2. IgD - recognition of the antigen by
B lymphocytes
◼ 3. IgE - allergic reactions
◼ 4. IgG - for complement fixation
◼ 5. IgM - also responsible for
complement fixation.
 Mechanism of Actions of
Antibodies

◼ 1. By direct actions
◼ 2. Through complement system
 Direct Actions of Antibodies

◼ i. Agglutination
◼ ii. Precipitation
◼ iii. Neutralization
◼ iv. Lysis
 Actions through
Complement System
◼ These are stronger than direct actions
◼ The plasma enzymes, are identified by
numbers from C1 to C9
◼ These enzymes are in inactive form
and are activated in three ways:
◼ a. Classical pathway
◼ b. Lectin pathway
◼ c. Alternate pathway.
 Classical pathway

◼ C1 binds with the antibodies and

triggers activation of other enzymes
in sequence.

◼ These enzymes or the byproducts

formed during these events produce
the following activities:
◼ i. Opsonization
◼ ii. Lysis
◼ iii. Chemotaxis
◼ iv. Agglutination
◼ v. Neutralization
◼ vi. Activation of mast cells and
basophils, which liberate histamine
 Lectin pathway

◼ Lectin pathway occurs when

mannose-binding lectin (MBL),
which is a serum protein binds with
mannose or fructose group on wall
of bacteria, fungi or virus.
 Alternate pathway

◼ Complementary system is also

activated by an alternate pathway.
◼ It is due to a protein in circulation
called factor I.
◼ It binds with polysaccharides present
in the cell membrane of the invading
organisms.
◼ This activates C3 & C5 which attack
the antigenic products
History of Blood Groups
and Blood Transfusions
•Experiments with blood transfusions
have been carried out for hundreds of
years. Many patients have died and it was
not until 1901, when the Austrian Karl
Landsteiner discovered human blood
groups, that blood transfusions became
safer.

• He found that mixing blood from two

individuals can lead to blood clumping.
The clumped RBCs can crack and cause
toxic reactions. This can be fatal.
 History of Blood Groups and
Blood Transfusions (Cont.)

• Karl Landsteiner discovered that blood

clumping was an immunological reaction
which occurs when the receiver of a blood
transfusion has antibodies against the donor
blood cells.

•Karl Landsteiner's work made it possible to

determine blood types and thus paved the
way for blood transfusions to be carried out
safely. For this discovery he was awarded the
Nobel Prize in Physiology or Medicine in
1930.
 What are the different blood
groups?
•The differences in human bld are
due to the presence or absence of
certain protein molecules called
antigens and antibodies.

•The antigens are located on the

surface of the RBCs and the
antibodies are in the blood
plasma.
 What are the different
blood groups?

• Individuals have different

types and combinations of
these molecules.

• The blood group you belong to

depends on what you have
inherited from your parents.
 What are the different blood
groups?

• There are more than 20

genetically determined blood
group systems known today
• The AB0 and Rhesus (Rh)
systems are the most important
ones used for blood
transfusions.
 What are the different
blood groups?

• Not all blood groups are

compatible with each other.
• Mixing incompatible blood
groups leads to blood clumping
or agglutination, which is
dangerous for individuals.
ABO blood grouping system

According to the
ABO blood typing
system there are
four different kinds
of blood types: A,
B, AB or O (null).
Antigens and Antibodies of
ABO Blood Types
 Why do individuals produce antibodies
to antigens they do not have?
• The "A“ and "B" antigens are also
produced by some other plants and
microorganisms. Thus, the body can
produce antibodies when exposed to
the plant or microbial antigens.

• These antibodies will also react

with human antigens of the same
kind whether introduced via a blood
transfusion or a tissue graft.
 ABO Inheritance and Genetics
• The ABO gene is autosomal (the gene is not on either
sex chromosomes)

• The ABO gene locus is located on the chromosome 9.

• A and B blood groups are dominant over the O blood
group
• A and B group genes are co-dominant
• Each person has two copies of genes coding for their
ABO blood group (one maternal and one paternal in
origin)
 AUTOSOMAL
CHROMOSOME
The alleles for Blood
group are in the same
A place on the B
chromosome 9. However
the genes have a
different code giving the
different blood group Mustafa

Sara

one alleles from Mustafa and one

from Sara.
 What do co-dominant genes
mean?
This meant that if a person inherited one A group
gene and one B group gene their red cells would
possess both the A and B blood group antigens.

These alleles were termed A ( which produced the A

antigen ),
B (which produced the B antigen) and O (which was
“non functional” and produced no A or B antigen)
 The ABO blood groups
• The most important in assuring a safe blood transfusion.
• The table shows the four ABO phenotypes ("blood groups") present
in the human population and the genotypes that give rise to them.

Blood Antigens
Antibodies in Serum Genotypes
Group on RBCs

A A Anti-B AA or AO
B B Anti-A BB or BO
AB A and B Neither AB
O Neither Anti-A and anti-B OO
 The Rhesus (Rh) System
• Another antigen to be considered is the Rh
antigen.
• Some of us have it, some of us don't.
• If it is present, the blood is RhD positive, if not
it's RhD negative.
• So, some people in group A will have it, and
will therefore be classed as A+ (or A positive).
• While the ones that don't, are A- (or A
negative).
• And so it goes for groups B, AB and O.
 The Rhesus (Rh) System (Cont.)

• Rh antigens are transmembrane proteins with loops

exposed at the surface of red blood cells.

• They appear to be used for the transport of carbon dioxide

and/or ammonia across the plasma membrane.

• They are named after the rhesus monkey in which they were
first discovered.

• RBCs that are "Rh positive" express antigen designated D.

• 85% of the population is RhD positive, the other 15% of the

population is running around with RhD negative blood.
 Rh Blood Group and Rh
Incompatibility
A person with Rh- blood does not have Rh
antibodies naturally in the blood plasma

Blood Alleles
Genotype
Type Produced

RR R
Rh positive
Rr R or r

Rh negative rr r
 Do you know which blood
group you belong to?
According to above blood grouping
systems, you can belong to either of
following 8 blood groups:
 Why is an Rh incompatibility so
dangerous when ABO incompatibility
is not during pregnancy?
• Most anti-A or anti-B antibodies are of the
IgM class (large molecules) and these do not
cross the placenta.

•In fact, an Rh−/type O mother carrying an

Rh+/type A, B, or AB foetus is resistant to
sensitisation to the Rh antigen.

•Her anti-A and anti-B antibodies destroy any

foetal cells that enter her blood before they
can elicit anti-Rh antibodies in her.
 Rh incompatibility during
pregnancy (cont.)
•This phenomenon has led to an effective
preventive measure to avoid Rh sensitisation.

•Shortly before each birth of an Rh+ baby, the

mother is given an injection of anti-Rh
antibodies (or Rhogam).

•These passively acquired antibodies destroy

any foetal cells that got into her circulation
before they can elicit an active immune
response in her.
 Blood transfusions – who can receive
blood from whom?

People with blood

group O are called
"universal
donors" and
people with blood
group AB are called
"universal
receivers."
Blood Antigens Antibodi Can give Can
Group es blood to receive
blood
from

AB
3%

A
42%

B
9%

O
46%
Blood Antigens Antibodi Can give Can
Group es blood to receive
blood
from

AB A and B None AB AB, A, B,

O

A A B A and AB A and O

B B A B and AB B and O

O None A and B AB, A, B, O

O
 Hemolytic Disease
◼ Rh blood group
◼ People whose RBCs have
the Rh antigen are Rh+
◼ People who lack the Rh
antigen are Rh-
◼ Normally, blood plasma
does not contain anti-RH
antibodies
◼ Hemolytic disease of the
newborn (HDN) – if blood
from Rh+ fetus contacts Rh-
mother during birth, anti-Rh
antibodies made
◼ Effect is on second Rh+ baby
 Typing Blood

◼ Single drops of blood

are mixed with
different antisera
◼ Agglutination with an
antisera indicates the
presence of that
antigen on the RBC