Blood

Swastika Hada
 General Properties of Whole Blood

• Fraction of body weight 8%

• Volume Female: 4-5 L, Male: 5-6 L
• Temperature 38° C (100.4° F)
• pH 7.35 - 7.45
• Viscosity (relative to water) Whole blood: 4.5-5.5, plasma: 2.0
• Osmolarity 280-300 mOsm/L
• Mean salinity (mainly NaCl) 0.85%
• Blood is composed of
plasma and blood cells

• Blood cells are also called

hemocytes
– Red blood cells : Erythrocytes
– White blood cells : Leukocytes
– Platelets : Thrombocytes.
 Blood plasma
• is a straw colored liquid component of blood that normally
holds the blood cells
• It makes up about 55% of the body's total blood volume.
• It is mostly water (up to 95% by volume),
• Contains :
– Inorganic molecules:Na+, Ca++, Cl-, HCO3-, K+, Mg++, Cu2+, PO43-,
Fe2+ , Fe3+
– Organic molecules :plasma protein, sugar, fats, enzymes,
hormones
– Plasma proteins: albumin, globulin,fibrinogen,prothrombin
– non protein nirtogenous substances: urea, uric acid, creatine,
creatinine, xanthine, hypoxantine
plasma without clotting factors is called serum.
Blood has three main functions
Transport
• Gases, namely oxygen (O2) and carbon dioxide (CO2), between the lungs
and rest of the body
• Nutrients from the digestive tract and storage sites to the rest of the body
• Waste products to be detoxified or removed by the liver and kidneys
• Hormones from the glands in which they are produced to their target cells
• Heat to the skin so as to help regulate body temperature

Protection
• Blood has several roles in inflammation:
• Leukocytes destroy invading microorganisms and cancer cells
• Antibodies and other proteins destroy pathogenic substances
• Platelet factors initiate blood clotting and help minimise blood loss

Regulation
• pH by interacting with acids and bases
• Water balance by transferring water to and from tissues
• In fetus, they are formed in liver and spleen
• In children, they are actively produced in marrow
cavities of all the bones
• In adults, blood cells are formed in the bone marrow
of the long bones (humerus, femur), vertebra,
sternum, ribs .
• Hematopoietic stem cells are capable of producing all
types of blood cells
 Erythrocytes (Red Blood Cells)
• Microscopic cells without nuclei
• Biconcave disc having mean
diameter 7.8 micrometer ,
thickness 2.5 micrometer at the
periphery
• Red cells normally make up
40-50% of the total blood volume.
– In average male: 4.5-6 million /mm3
– In average female:4.0-5.5 million /mm3
• Red cells have ability to
concentrate hemoglobin
– Male: 15 gm/100ml cells
– Female : 14 gm /100ml cells
• Each gram of hemoglobin contains
1.34 ml of oxygen
 Hemoglobin
• Hemoglobin is a globular
protein consisting of 4 subunits,
each subunit contains the
pigment heme conjugated with
globin polypeptide.

• There are 2 pair of polypeptide in each hemoglobin

molecule
• In adult hemoglobin (HbA):α2β2
– 25% of adult Hb is A2 : α2δ2
– Small amount of Hb A has glucose attached to its terminal :
HbA1c

• Fetal hemoglobin (HbF): α2γ2

• Heme is red, iron containing moiety in which O2 bind
to Fe++ to form oxyhemoglobin
• When CO2 binds to the Hb carbaminohemoglobin
• When CO binds to Hb --> carboxyhemoglobin
• Presence of ferric iron methmoglobin
• Myoglobin is a hemeprotein present in heart and
skeletal muscle, Functions both as a reservoir and
carrier of oxygen
 .
Hematopoiesis
• refers to whole blood cell production
• All blood cells, including red & white, are produced in
red bone marrow
– arise from the same type of stem cell, the hematopoietic
stem cell
• Blood forming tissues, which produce the RBCs, WBCs &
platelets, are usually divided into two types.
• Myleoid tissue
– Means red bone marrow
– It produces the RBCs, neutrophils, eosinophils, basophils,
monocytes & the platelets
• Lymphoid tissue
– includes the lymph nodes, the thymus and the spleen
• Erythropoiesis
• refers specifically to red blood cell production
 Genesis of RBC
o Blood cell begin their life in bone
marrow from pluripotential
hematopoietic stem cell which
differentiates to form committed
stem cells
o The first cell that can be identified
as belonging to RBC is
proerythroblast which divides
many times to form mature RBC
o Basophil erythroblast has very few
hemoglobin but its concentration
increases and nucleus goes on
degrading in succeeding
generations
o Reticulocyte stage passes from
bone marrow to capillaries
 Regulation of RBC production
Tissue oxygenation:
• Any condition that cause quantity of oxygen
transported to tissue to decrease causes increase
the rate of RBC production
• Anemia, high altitude, lung disease, cardiac failure
Erythropoietin:
• Formed mostly in kidney and some in liver
• Stimulate production of proerythroblast from
hematopoietic stem cell in the bone marrow
• Maturation of RBC requires vitamin B 12 and folic
acid
Life span and destruction of RBC
• In the circulation, RBC survive for
120 days
• As the cell becomes older they
becomes fragile and rupture when
passes through tight spot in the
circulation.
• Most of them are destroyed in
spleen and liver
• The released hemoglobin is
phagocytosed by macrophages
• Released iron and protein are
recirculated
• Heme is converted into bilirubin and
excreted in bile
Anemia: deficiency of hemoglobin in blood
• Blood loss anemia: occur due to rapid hemorrhage
– In chronic blood loss , person cannot absorb enough iron
from the intestine to form Hb as rapidly as it is formed
– Red cells produced are smaller and condition is called
microcytic hypochromic anemia
• Aplastic anemia: due to lack of functioning bone
marrow
– Bone marrow might get damaged due to exposure to
radiations, chemicals and drug, autoimmune disorder
• Megaloblastic anemia: lack of vit B12 and folic acid
causes slow reproduction of RBC resulting large , odd
shaped cells
– Caused due to atrophy of stomach mucosa and gastrectomy
– Megaloblastic anemia caused by vitamin B-12 deficiency is
referred to as pernicious anemia.
Hemolytic anemia: rate of cell destruction is faster than
their formation
• Sickle cell anemia :having faulty beta chain
– When this Hb is exposed to low oxygen concentration, it
precipitate into long crystal inside RBC and gives sickle shape
– Precipitated Hb also damage the red cell membrane and
makes It fragile
• Hereditary spherocytosis
– Red cells are spherical so cannot withstand compression
force
– Easily rupture on passing through vascular bed
– caused by a molecular defect in one or more of the proteins
of the red blood cell cytoskeleton
• Erythroblastosis fetalis
– Rh incompatibility makes the Rh positive cells fragile leading
to rapid RBC rupture in child
Polycythemia: increase in number of red cells
– High altitude
– Cardiac failure
– Polycythemia vera (Erythremia) :polycythemia of unknown cause
 Blood types
• The membrane of human red cells contains variety
of blood group antigens called agglutinogens.
• The most important and best known are A and B
antigen
• Depending upon presence and absence of these 2
antigens four major blood types can be classified
(ABO system )
– Presence of A agglutinogen: blood type is A
– Presence of B agglutinogen: blood type is B
– Presence of both A and B agglutinogen : blood type AB
– Absence of both A and B agglutinogen : blood type O
Agglutinins (antibodies)
• When type A agglutinogen is not present in the
persons RBC, antibodies known as anti A
agglutinins are developed in the plasma
• When type B agglutinogen is not present in the
persons RBC, antibodies known as anti B
agglutinins are developed in the plasma
• Type O contains no agglutinogen but contains
both anti A and anti B agglutinins
• Type AB blood contains both A and B
agglutinogen but no agglutinins
 Rh blood types
• There are 6 common types of Rh antigen (C,D,E,c,d,e)
which are called Rh factors
• D antigent is widely prevalent in population and are
considerably more antigenic
• Blood group having D antigen is called Rh positive blood
and that without Rh antigen is called Rh negative blood
 Blood typing
• Before transfusing blood to the person, it is
necessary to determine the blood types of donor
and recipient
• Clumping of blood occurs due to antigen –antibody
reaction , if the blood types does not match:
agglutination
 Characteristics of Rh transfusion reaction
• If an Rh negative person has never before been
exposed to Rh positive blood , transfusion of Rh
positive blood to that person will cause no
immediate reaction
• However, anti Rh antibodies can develop in
sufficient quantity in next 2-4 weeks causing
agglutination of those transfused cells that are
still circulating in the blood.
• On subsequent transfusion of Rh positive blood
into same person , the reaction is greatly
enhanced and can be immediate and severe
 Erythroblastosis fetalis
• Disease of foetus and newborn characterized by
agglutination and phagocytosis of fetus’s RBC
• When Rh- mother carries Rh+ fetus, small amount of
fetal blood leaks into maternal circulation at the time of
delivery
• Mother develops significant titer of agglutinins during
post partum period
• During the next pregnancy, if the mother carries Rh+
fetus again , the mother’s agglutinin cross the placenta
and causes hemolysis of fetal blood
• If the hemolysis is severe, infant may die in uterus or
may develop anemia, severe jaundice and edema
(hydrops fetalis)
• Bilirubin rarely penetrate brain , but in case of
erythroblastosis, bilirubin gets deposited in the basal
ganglia leading to neurological syndrome called
kernicterus
Leukocytes
• Formed partially in bone marrow and partially in
lymph tissue
• 4000-11,000 WBCs per microliter of blood
• Prevents disease in the body by
– Destroying invading bacteria
– Forming antibodies and sensitized lymphocytes
• 5 types of WBC can be classified into 2 groups
Granulocytes: Agranulocytes
– Neutrophils -Lymphocytes
– Eosinophils -Monocytes
– Basophils
• Neutrophils:
– constituting 60-70% of the circulating leukocytes
– Diameter varies from 10-14μm
– Nucleus are of various shapes, usually contains 2-5 lobes
– Contains fine pink granules
– Are phagocytic in nature, contains alkaline phosphatase,
defensin, lysosyme, collagenase, sulphatases etc
– They are usually first responders to microbial infection
• Eosinophils:
– compose about 2-4% of the WBC
– Diameter varies from 10-14μm
– Nucleus usually has 2 lobes (headphone shaped)
– Cytoplasm is packed with refractile brick red coarse granules
– It rises in response to allergies, parasitic infections and
disease of the spleen and central nervous system
– They are rare in the blood, but numerous in the mucous
membranes of the respiratory, digestive, and lower urinary
tracts
– Granules contains major basic protein, eosinophil cationic
protein, eosinophil peroxidase, eosinophil-derived
neurotoxin
• Basophil:
– Rarest leucocytes representing
about 0.5 to 1% of
circulating WBC
– Nucleus occupy greater
portion of cell, irregular in
shape
– Nucleus is obscured by deeply
stained blue-black granules
– Contains heparin, histamine,
• Theyserotonin
are responsible for inflammatory reactions during
immune response, acute and chronic allergic diseases,
like anaphylaxis, asthma, atopic dermatitis and hay
fever.
• Monocytes:
– Monocytes compose 2% to 10% of all leukocytes
– Largest leucocyte 12-25μm
– Large kidney shaped, ecentrically located nucleus
– Phagocytic , protects body from bacteria, virus, fungi
and parasites
– Remain in circulation for few hours and migrate to
organs and transform to macrophages
Lymphocytes
• Small lymphocytes:
– Size same or slightly bigger than RBC
– Cell are mainly occupied by purple
stained round nucleus
• Large lymphocyte:
– Same size or slightly larger than
neutrophil
– Nucleus occupy 90 % of the cell
– Cytoplasm is clear and navy blue in color
• Lymphocytes mediate immunologic
response
• Functionally it can be divided into
– B lymphocyte : mediate humoral
immunity
– T lymphocytes : mediate cell mediated
immunity
• T and B lymphocytes are morphologically
indistinguishable but can be identified by
markers on their cell membrane
• B cells differentiate into plasma cells and
memory B cells
• There are 3 major types of T cells :
– Cytotoxic T cells(CD8 cells) : destroy transplanted
and other foreign cells
– Helper T cells(CD4 cells): secrete cytokines that
enhane the activity of macrophages, NK cells, B cells,
cytotoxic T cells, other Helper Tcells
– Supressor T cells: secrete cytokines that suppress the
activity of cytotoxic T cells, Helper T cells , B cell
• Life span of granulocytes after release from the
bone marrow is 4-8 hours in blood and another
4-5 days in tissue
• Monocytes have short transit time of 10-20 hours
in blood . Once in the tissue , they swell to much
larger size to become tissue macrophage and live
for months unless destroyed while performing
phagocytic function
• Lymphocytes enter circulatory system from
lymphoid tissues and circulate through the tissue.
They have life span of weeks or months depends
on body’s need
 Neutrophil and macrophages defend against infection
• plays important role in inflammation
• They enter the tissue spaces by
diapedesis (squeeze through the pores of
blood capillaries)

• They move through tissue spaces by

amoeboid motion

• They are attracted to the inflamed tissue

by chemotaxis
– Bacterial or viral toxins
– Degenerative product of inflamed tissue
– Complement complex

• Bacteria are coated by antibodies (opsonins) and make it suitable for

phagocytosis, a process is called opsonization

• Coated bacteria are then phagocytosed by endocytosis (Phagocytosis)

 Reticuloendothelial system
• Includes the total of monocytes,
mobile macrophage and tissue
macrophages

• Is generalized phagocytic
system specially located in
those tissues where large
quantities of particles , toxins
should be destroyed

• Tissue macrophages in skin and

subcutaneous tissue:
Histocytes
• In liver : Kupffer cells
• In lungs: alveolar macrophages
• In brain : microglia
 Immunity
• Ability to resist all types of organism or toxins that
tend to damage the body
Innate immunity: It includes
– Phagocytosis by reticuloendothelial system
– Destruction of swallowed organism by acid and digestive
enzymes in stomach
– Resistance of skin to the invading organism
– Lysosomes, complements, natural killer cells, WBCs
Adaptive (aquired) immunity
• Do not develop until after the body is first attacked
by pathogen
– Humoral immunity ( B cell mediated)
– Cell mediated immunity ( T cell mediated)
• Acquired immunity is the extremely powerful
immunity against individual invading agents such
as bacteria, virus, toxins

• Acquired immunity do not develop until after the

invasion of foreign organism or toxins

• Any toxins or organism contains one or more

specific compounds (protein or polysaccharides)
are called antigen

• Lymphocytes are responsible for acquired

immunity
Humoral immunity
• Before exposure to specific antigen , B lymphocytes
remains dormant in lymphoid tissue
• On entry of antigen, macrophage phagocytize the
antigen and present to the B lymphocyte
• B lymphocyte(lymphoblast) specific for antigen
differentiate to plasmablast (precursor of plasma
cells)
• Plasmablast divides rapidly to give plasma cells
• Mature plasma cells produces immunoglobulin
• A few lymphoblast do not form plasma cells , but
form its clone that circulate in the body: memory
cells
• When the same antigen is exposed for second time ,
these memory cells are responsible to produce rapid
immune reaction
Antibodies acts by
• Direct action to invading agents
– Agglutination
– Precipitation
– Neutralization
– Lysis

• Activating complement system

 Cell mediated immunity
• After exposure to antigen specific T lymphocytes are
produced along with B lymphocyte
• Instead of releasing antibodies, whole T cells are involved
in the reaction
• T lymphocyte respond to antigen only when they are
bound to MHC protein (major histocompatibility complex)
present in antigen presenting cells ( macrophages,
dendritic cells)
• Types of MHC protein:
• MHC-I:
– present in all the nucleated cells
– Present antigen to cytotoxic T cell
• MHC-II:
– present in APCs including B and T cells
– Present antigen to Helper T cells
• APCs ingest and partially digest an antigen then
present the part of the antigen along with MHC II
• Immune synapse form with CD4 T cells, which
activates to produce IL2
• IL2 act in autocrine fashion to cause the cell to
multiply
• Activated CD4 cell may promote B cell activation
to produce plasma cells or activate cytotoxic CD 8
cells
• Cytotoxic T cell directly kills the microorganism by
producing hole forming protein called perforin
• It releases cytotoxic substances through the pore
and kills the cell
Leukopenia: Condition in which bone marrow produce
very few WBCs
Irridation of body by X rays and gamma rays is likely to
cause aplasia of bone marrow
Within 2 days after bone marrow stops producing
WBCs ulcer appears in GIT, respiratory infections

Leukemias: Uncontrolled production of WBCs

characterized by increased number of abnormal
WBCs in the circulation
Almost all leukemia spread to lymph nodes, liver and
other vascular regions
Common effects are development of infection,
anemias, thrombocytopenias due to displacement of
normal bone marrow by leukemic cells
Platelets
• Are only about 20% of the diameter of red blood cells.
• Normal concentration: 150,000-400,000/mm3
• Cytoplasm contains actin, myosin, thrombosthenin ,
residue of golgi apparatus and ER ,mitochondria,
• Granules contains serotonin , Ca++, ADP, insulin-like
growth factor 1, platelet-derived growth factors, platelet
factor 4
• Are anucleated, half life in blood is 8-12 days
• Formation is regulated by thrombopoietin
• The main function is to prevent bleeding
• Platelets are actually not
true cells but merely
circulating fragments of
cells called
megakaryocytes
• As megakaryocytes
develop into giant cells,
they undergo a process
of fragmentation that
results in the release of
over 1,000 platelets per
megakaryocyte.
 Hemostasis
• Means prevention of blood loss
• Whenever the vessel is severed or ruptured,
hemostasis is achieved by
1. Vascular constriction
2. Formation of platelet plug
3. Formation of blood clot
4. Growth of fibrous tissue into blood clot to
close the hole permanently
Vascular constriction
• Immediately after the blood vessel has been cut, the
trauma to the vessel cause the smooth muscle in the
wall to contract
• The contraction result from:
– local myogenic spasm
– Autocoid factors (eicosanoids, angiotensin,
neurotensin, NO, histamine, serotonin )
– Nervous reflex
– The spasm can last for many minutes or even hours
Formation of platelet plug
• When platelet comes in contact with collagen of
damaged vascular surface, they begins to swell
• Becomes sticky so that they adhere to collagen in
tissue and to a protein called von willebrand factor
that leaks from traumatize tissue to plasma
• Produces pseudopodes, contractile proteins contract
forcefully and cause release its granules
• Produce ADP and thromboxane A2 which in turn
activates other paltelets and cause them to adhere
to original activated platelets
• Progressively accumulates more and more platelets
to form platelet plug
Formation of blood clot
• The loose aggregration of platelet in the temporary
plug is converted into definitive plug
Conversion of prothrombin to thrombin
• Prothrombin activator is formed as a result of rupture
of blood vessel
• In presence of Ca++ it causes conversion of prothrombin
to thrombin [prothrombin is formed by liver n presence of
vitamin K ]
• The thrombin causes polymerization of fibrinogen
molecule into fibrin that entanfles with RBC, platelets
and plasma to form blood clot
• The strong cross linkage between the fibrin molecules is
formed by fibrin stabilizing factor(released from platelets
entrapped in the clot) that strengthens the fibrin meshwork.
Prothrombin can be activated by 2 pathways
Intrinsic pathways
Extrinsic pathways
Clotting factors :
Extrinsic pathway
• The extrinsic pathway begins with traumatize
extravascular tissue that comes in contact with
blood
• Traumatize tissue release complex of several
factors called tissue factors or thromboplastin . This
factor is composed of phospholipids and
lipoproteins
• Tissue thromboplastin complex with factor VII in
presence of Ca++ to activate factor X into Xa
• Xa combine immediately with phospholipid as well
as factor V to form prothrombin activator
Intrinsic pathway
• Intrinsic pathway begins with activation of factor
XII Which is catalyze by high molecular weight
kininogen and kallikrein due to exposure of
blood to collagen from traumatize blood vessel
wall or exposing the blood to glass
• Intrinsic pathway is slow to proceed requiring 1-5
minutes to cause clotting while clotting can occur
with extrinsic pathway as little as 15 seconds
• Ca++ is required for promotion and acceleration
of all the blood clotting reaction except for first 2
steps in intrinsic pathway
 Clot retraction
• Within a few minutes after a clot is formed, it begins to
contract and expresses most of the fluid from within
20-60 minutes.
• The fluid expressed is called serum because all its
fibrinogen and most of the clotting factors have been
removed.
• Platelets are necessary for clot retraction to occur
• Platelet entrapped in the clot release fibrin stabilizing
factor which increase the bond between the adjacent
fibrin fiber.
• It also activates the contractile protein that helps to
compress the fibrin meshwork into smaller mass
• As the clot retracts, the edges of the broken vessels are
pulled together contributing further to hemostasis .
Preventing blood clotting in normal vascular system
• Smoothness of endothelial cell surface
• Layer of glycocalx on the surface of endothelium
that repels clotting factors and platelets
• Protein bound with endothelium called
thrombomodulin that binds with thrombin
– Thrombomodulin-thrombin complex activates plasma
protein called protein C that acts as anticoagulant by
inactivating factor V and VIII
• Antithrombin III blocks the activity of clotting
factors .
– Its binding with clotting factor is facilitated by
heparin(naturally occurring anticoagulant)
– Clotting factors that are inhibited are IX,X, XI, XII
 Lysis of blood clot
• Plasma protein contains Plasminogen that
when activated becomes Plasmin( fibrinolysin)
• Plasmin is a proteolytic enzyme that digest
fibrin and other coagulant such as fibrinogen,
factor V, VIII, XIII and prothrombin.
• Large amount of plasminogen is trapped in
the clot which get activated by tissue
plasminogen activator(t-PA) into plasmin
which in turn remove unnecessary blood clot
Positive feedback of clot formation
• Thrombin has direct proteolytic effect on prothrombin
itself, tending to convert it into still more thrombin
• Once a critical amount of thrombin is formed , a positive
feedback develops that cause still more blood clotting and
more thrombin to form
• Blood clot continues to grow until blood leakage ceases.
Conditions that causes excessive bleeding in
humans
• Hemophilia: exclusively occur in males
– Haemophilia A : Caused by abnormality or deficiency
of factor VIII
• Thrombocytopenia:
– Presence of very low number of platelets
– Bleeding is from small Venules and capillaries
– Skin of such parson displays small purplish blotches
called purpura
• Vitamin K deficiency:
– Due to liver cirrhosis, hepatitis
– Causes decreased formation of factor VII, XI, X and
prothrombin
Thrombocytosis: condition when
platelet count is increased in
blood
Thrombocytopenia: condition in
which platelet count is
decreased in the blood
• Caused by impaired production
or increased destruction
• Results in purpura characterized
by easy bruisability and multiple
subcutaneous hemorrhage
Prevention of blood coagulation outside of the body
• Heparin
• Oxalate: decrease ionic calcium level in blood
• Sodium, potassium, ammonium citrate: combine
with calcium to form unionize calcium
• Blood collected in siliconized container does not clot
for 1 hour or more because silicon prevents contact
activation of platelets and factor XII
• EDTA: chelation of calcium ion