BLOOD PHYSIOLOGY

• Blood is a red blood fluid that circulates through the vascular system
• It carries nutrients and waste products throughout the body

PROPERTIES OF BLOOD
• It is red in colour,
• Arterial blood is scarlet red while venous blood is purple red because of more Co2
• Average volume of blood in a normal human is 5l
• In newborns it is 450ml and increases through growth and reaches 5L at the tine of puberty
• Blood in females is 4,5 L
• It is 8% of body weight in normal young healthy adult weighing about 70kg
• It is also 5 times more viscous than water
• pH range from 7.35 to 7.45 (slightly alkaline).
• Slightly warmer than body temperature 100.4 oF.

COMPOSITION OF BLOOD
• It is composed of RBC,WBC,PLATELETS
HAEMATOCRIT VALUE / PCV
• It is the volume of RBC in the blood expressed by percentage
• The normal value is 45%
• (47% in male, 42% in female)
• Between the RBC and plamsa cells, a thin layer of white buffy coat is present
• The white buffy coat is formed by the aggregation of WBC and platlets
• When blood is collected in an haematocrit and centrifuged for 30 mins at the speed of 3000rpm, RBC settles leaving blood
plasma at the top.
• Plasma forms 55% while RBC forms 45%
PLASMA
• It is a straw liquid part of blood
• It contains 91-92% of water and 8% of solids and gases

SERUM
• Clear fluid that oozes from blood clot
• When blood clot happens fibrinogen is converted to fibrin and blood cells are trapped in this forming blood clot
• After 45 minutes serum oozes out of the blood clot
• Serum is different from plasma by only the absense of fibrinogen
• Fibrinogen is absent in serum because it is converted to fibrin during blood clotting

FUNCTION OF BLOOD
• Nutritive
• Respiratory
• Excretory
• Transport of hormones and enzymes
• Water balance E.T.c
PLASMA PROTEINS

SEPERATION OF PLASMA PROTEINS:

• PRECIPITATION METHOD: Proteins in serum are seperated into albumins and globulins
• SALTING OUT METHOD: Serum globulin is seperated into euglobulin and pseudoglobulin
• ELECTROPHORETIC METHOD: Proteins are seperated into albumin (55%), alphaglobulin (13%),beta globulin (13%), gamma globulin
(11%) and fibrinogen (7%)
• COHN"S FRACTIONAL PRECIPITATION METHOD: Plasma proteins are seperated into albumin and different fractions of globulin
• ULTRACENTRIFUGATION: Albumin,GLOBULIN and FIBRINOGEN are seperated and molecular weight also is determined
• GEL FILTERATION CHROMATOGRAPHY: All types of plamsa proteins are seperated
• IMMUNOELECTROPHORETIC: Quantitative measurement of proteins is done

PROPERTIES OF PLASMA PROTEINS:

MOLECULAR WEIGHT: Molecular weight of fibrinogen is higher than that of the others
ONCOTIC PRESSURE: Plasma proteins exert oncotic pressure on the blood, albumin contributes the most
SPECIFIC GRAVITY: the specific gravity of proteins is 1.026
BUFFER ACTION: Proteins have 1/6 action of the buffering ability of blood

ORIGIN OF PLASMA PROTEINS:

IN EMBRYO-
• Plasma proteins are synthensized first by mesenchyme cells
• Albumin is synthensized first and other proteins are synthensized later
IN ADULTS
 • They are synthensized mainly from the reticuloendothelial cells of liver
• The plasma proteins are synthensized from spleen,bone marrow
• Gamma globulin is synthensized from Blymphocytes

EXAMPLE OF PLASMA PROTEINS

ALBUMIN
• Most abundant (55-64%)
• 4-5g/100mls of blood (3.5-5g/dl)
• The smallest size with molecular weight of 69000-70,000
• Total exchangeable albumin is 4-5g/kg body weight
• 38.45% intravascular
• 10% of exchangeable pool degraded daily
• Replacement comes from hepatic cells producing 200 to 400mg/kg/day
• Synthesis decreases in fasting and increase in nephrosis because of loss in urine
GLOBULIN
• Form about 20% of total plasma protein
• Amount in circulating blood is about 2-3g/100ml of blood
• Occurs in various form
o - α- globulin (150,000-160,000).Transport retinol and Thyroxine
o - Y-globulin (150,000 – 900,000). E.g. iron transporting protein called trasferrin.
o - β- globulin (90,000) e.g. antibodies
• Albumin-globulin ratio = 2:1
FIBRINOGEN
• Only soluble plasma protein
• Molecular weight = 350,000
• Amount in circulation = 0.15 – 0.3g/100ml
• Converted to fibrin as blood coagulates

COMPONENT PERCENTAGE (%)

Water 92
Proteins 6-8
Salts 0.8
Lipids 0.6
Glucose (blood glucose) 0.1

FUNCTION OF PLASMA PROTEINS:

• Coagulation of blood
• Defense mechanism- Gamma globulins or immunoglobulins
• Transport mevhanism
• Acid base balance regulation
• Viscosity of blood- Albumin helps in this the most
• Transport of blood- Alpha and beta globulins help in transport of metals, Albumin, alpha and beta globulin helps in the
transport of enzymes
• Globulin and fibrinogen helps to accelerate the tendeency of roleaux formation by cells used in ESR Rate
• Production of trephone substance used for norishment of tissue cells
• They are the last resort of energy
ONCOTIC PRESSURE MAINTENANCE
• At capillary level, most of the substances are exchanged between blood and tissues
• Because of plasma proteins cannot pass through cell membrane easily and remain in blood
• Albumin generates 70% of blood pressure
• Oncotic pressure is 25mmhg
• Concentration of albumin is more than other plasma proteins
• Globulin is the next and fibrinogen
RED BLOOD CELS

• Erythrocytes are also known as red blood cells.

• Mature red blood cell is non nucleated & has a biconcave shape.
• It has a diameter of 8.5micrometer, edge thickness of 2.5 & center thickness of 1.5 micrometer.
• There are about 5 million/mm3 of blood with variation between the two sexes.
• Male 4.5-6 & female 4.3-4.5 million cells/mm3 of blood.
The variation is due to the following;
1. Gonadal hormone testosterone in male which is a red blood cell differentiation inducer.
2. Greater mass musculature in male requires more oxygen supply.

OTHER VARIATIONS
• Increase in RBC count is known as polythemia
 • RBC count also decreases during sleep and pregnancy
• People in high altitutude have high RBC count, hypoxia stimulates the kidney to produce erythropoetin which inturn
stimulates bone marrow to produce more RBC
• RBC Count increases after meal,during emotional anxiety e.t.c
• The shape of the red blood cell provides maximum surface area for the small volume of the cell and large diffusion surface
for passage of gases.
• It also allows for squeezing of red blood cell when passing through narrowed vessels without damage to the integrity of the
cell.
• Glucose-6-phosphate dehydrogenase is present on the membrane helping to utilize glucoce, O2 and ATP.
• Life span is 120 days.
• The count is higher in new born (6mmillion/mm3) than in an adult.
• Its production is stimulated by hypoxia with the release of erythropoietin (EPO) 85% from kidney % 15% from liver.
• RBC Count decreases during pregnancy because of increase in ECF Volume which increases plasma volume leading to
haemodilution

STRUTURE OF RBC
▪ They are non nucleated, only camel has nucleated RBC
▪ They also lack DNA
▪ Energy for RBC is gotten from glycolytic process since it lacks mitochondria
▪ RBC has a special type of cytoskeleton made up of actrin and spectrin and they are anchored to transmembrane proteins with
anktrin
▪ Absence of spectrin leads to hereditary sperocytosis in which cell is deformed and easily degraded
▪
FATE OF RBC
• Old RBC are destroyed by macrophages present in spleen and liver
• When rbc become older their cell membrane becomes more fragile
• Young RBC can pass through capillaries easily but because of fragile nature, older cells are destroyed attempting to pass
through
• Destruction occurs mainly in capillaries of red pulp of spleen because their diameter is very small
• Destroyed RBC's are fragmented and haemoglobin is released from fragmented parts
• haemoglobin is picked up by macrophages in the liver e.g kupffer cells, spleen and bone marrow
• It is degraded into iron,,globin and porphyin
• Iron combines with protein called apoferritin to form ferritin
• Globin enters the proteins depot for later use
• Porphrin is degraded into bilirubin and excreted by liver through bile
• Daily 10% RBCS are destroyed
USES OF RBC
• Transport of Oxygen
• Transport of CO2 to lungs (30% carbhaemoglobin, 63% Carbonic anhydrase)
• Buffering action
• Blood group determination: They carry blood group antigens
PHYSIOLOGICAL VARIATIONS
• INCREASE IN RBC COUNT- PATHOLOGICAL POLYCYTHERMIA
▪ Abnormal increase in RBC count above 7 million/cu mm of blood
PRIMARY POLYCYTHERMIA
▪ It is characterized by persistent increase in RBC count above 14 million/cu mm of blood
▪ It is associated with WBC above 24 million/cu mm of blood
▪ Occurs in myeloproliferative disorder
SECONDARY POLYCYTHERMIA
▪ Secondary pathological conditions like emphyesema, conenital heart disease and chemical poinsoning
▪ This would lead to hypoxia which stimulates erythropoetin which stimulates bone marrow, which stimulates RBC production
Decrease in RBC count is known as anaemia

VARIATIONS IN SIZE OF RBC

▪ Size of RBC in venous blood is larger than those in arterial blood
▪ Smaller RBC- Microcytes and are present in iron deficiency anemia, prolonged forced breathing and increased osmotic
pressure
▪ Larger RBC- Macrocytes and are present in megaloblastic anemia and decreased osmotic pressure in blood
▪ Anisocytes- Different Sizes and occurs in pernicious anemia
VARIATION IN RBC SHAPE
▪ Crenation: Shrinkage as in hypertonic solutions
▪ Spherocytosis: Globular form in hypertonic conditions
▪ Elliptocytosis: Elliptical shape as in certain anemia types
▪ Sickle Cell: Cresentic shape as in sickle cell
▪ Poikilocytosis: Deformed Cell membrane
VARIATION IN RBC STRUCTURE
PUNCTATE BASOPHILISM
▪ Striated appearance of RBCs by the presence of dots of basophilic materials (porphyrin) is called punctate basophilism.
▪ It occurs in conditions like lead poisoning.
RING IN RED BLOOD CELLS
▪ Ring or twisted strands of basophilic material appear in the periphery of the RBCs.
▪ This is also called the Goblet ring.
▪ This appears in the RBCs in certain types of anemia.
HOWELL-JOLLY BODIES
▪ In certain types of anemia, some nuclear fragments are present in the ectoplasm of the RBCs.
▪ These nuclear fragments are called Howell-Jolly bodies.

ERYTHROPOESIS
▪ This is the origin development and maturity of erythrocytes
▪ Haematopoesis is the process of origin and maturation of all blood cells

SITE OF ERYTHROPOESIS
▪ Occurs in 3 stages in foetal life
MESOBLASTIC STAGE: First 2 months RBC are produced in the mesenchyme cells of yolk sac
HEPATIC STAGE: for the 3rd month liver is the main organ that produces RBC, Spleen and other organs are also involved in
erythropoesis
MYELOID STAGE: Last 3 months of intrauterine life, RBC are produced from bone marrow and liver
▪ In newborn babies and adults, they are produced only from the bone marrow
▪ In adults, they are also produced from the bone marrow
UP TO 20 YEARS: RBC are produced from the bone marrow of all bones
AFTER 20 YEARS: Shaft loses erythropetic function and becomes yellow marrow because of fat deposition, so RBCS are produced
from membranous bones and from the end of long bones
▪ 75% of the bone marrow is involved in leukocytes production and 25% is involved in erythrocytes production

PROCESS OF ERYTHROPOESIS
HEMATOPOIETIC STEM CELLS:
▪ Primary cells capable of self renewal and diffrentitaion into specialized cells
▪ They are found in the bone marrow and they give rise to blood cells
▪ They are pluripotent
▪ In the early stage they are not committed but they later become comitted and it’s the commited that gives rise to a group of
RBC
TYPES OF COMMITTED PHSC
LYMPOID STEM CELLS: They give rise to Lymphocytes and natural killer cells
COLONY FORMING BLASTOCYTES: When grown in cultures they form colonies i.e colony forming blastocytes
DIFFERENT UNITS OF COLONY FORMING CELLS
CFUE-Erythrocytes
CFUGM- Granulocyes (Neutrophils,Baspohils and esinophils) and monocytes
CFUM- Platelets

CHANGES DURING ERYTHROPOESIS

▪ Reduction in cell size from 25-7.2 micrometer
▪ Disappearance of nucleoli and nucleus
▪ Appearance of haemoglobin
▪ Change in staining properties of cytoplasm

STAGES DURING ERYTHROPOESIS

PROERYTHROBLAST: PRONORMOBLAST
▪ First cell developed from CFU E
▪ Diameter of about 20 micrometers
▪ Does not contain haemoglobin
▪ Cytoplasm is basophlic in nature
▪ Contains large nucleus with 2 or more nuclei
▪ Multiplies severaly and forms cell of next stage

EARLY NORMOBLAST
▪ Nucleoli in nucleus disappears
▪ Becomes small with a diameter of 15 micrometer
▪ Chromatin network gets condensed and becomes dense
▪ Also known as basophillic erythroblast

INTERMEDIATE NORMOBLAST
▪ Smaller diameter of 10-12
▪ Nucleus is still present but further condensation
▪ Haemoglobin starts appearing
▪ Cytoplasm is also basophilic
▪ Because of haemoglobin it stains with acid as well as base
▪ Also called polychromatic erythroblast

LATE NORMOBLAST
▪ Diameter of cell decreases for about 8-10
▪ Nucleus becomes small with condensed chormatin network and is known as ink spot nucleus
▪ Quantity of haemoglobin almost increases and cytoplasm becomes almost acidophilic
▪ Cell becomes orthochromatic erythroblast
▪ In the final stage nucleus disintegrates and disappears, this process is known as pyknosis
▪ Final remnant is extruded and develops into reticulocyte

RETICULOCYE
▪ It is the immature RBC AND LARGER than the mature RBC
▪ It contains reticular network formed by remnants of disintegrated organelles
▪ Reticulum stains with supravital stain
▪ In babies it is 2-6% count of RBC's and count decreases after first week of birth
▪ Number increases when production and release of RBC increase
▪ Reticulocyte is basophillic due to the remnants of cell organelles
▪ Cells enter blood capillaries through the capillary membrane from site of production by diapedesis

MATURED ERYTHROCYTE
▪ Reticularr network disappears
▪ It is acidophillic
▪ Cells becomes matured and attains biconcave shape
▪ Diameter of cell reduces to 7.2
▪ It takes 7 days for development form proerythroblast to RBC
▪ It takes 5 days to get to reticulocye stage
▪ Reticulocyte takes 2 days to become matured RBC

FACTORS NECESSARY FOR ERYTHROPESIS

STIMULATING FACTORS

HYPOXIA
▪ Reduced avaliability of oxygen
▪ Major stimulating factor
▪ Stimulates erytrhopoesis by inducing secretion of erythropoetin in the kidney
▪ It involves Hypoxia INDUCTIBLE factor
▪ HYPOXIA INDUCIBLE FACTOR is a transcription factor used to activate the transcription of genes to produce substances
including erythropoeitin from proteins

ERYTHROPOIETIN
▪ Secreted by peritubular capillaries of kidney
▪ Hypoxia is its stimulant
ACTIONS OF ERYTHROPOETIN
▪ It takes 4-5 days
▪ Production of erythroblast from CFUE
▪ Development of proeryhtroblast into mature RBC's
▪ Release of mature RBC into blood'

THROXINE
▪ Accelerates erythropoesis
▪ Polycthemia is common

ROLE OF SEX HORMONES

▪ Estrogen suppresses erythropoiesis

HEMATOPOIETIC GROWTH FACTORS

▪ They are interlukins and stem cell factors
▪ Interlukins involved in erythropoiesis are:
Interleukin-3 secreted by T cells
Interleukin-6 secreted by T cells,endothelial cells and macrophages
Interleukin-11 secreted by osteoblast

VITAMINS
▪ Vit B3,B6,C,D,E

MATURATION FACTORS
VITAMIN B12: It is called an extrinsic factor since it is obtained from diet
▪ It is mostly stored in the liver
▪ Its absorption reqiures the presence of intrinsic factor of castle
▪ Deficiency of vitamin B12 causes pernicious anaemia

INTRINSIC FACTOR OF CASTLE:

▪ Produced by parietal cells of gastric glands
▪ Essential for the production of vitamin B2
▪ Deficiency occurs in severe gastritis, ulcer and gastrectomy

HEMATINIC PRINCIPLE
It is a maturation factor and also called anti-aemia princple
 FOLIC ACID
▪ In the absence of DNA, DNA synthesis decreases and it results in failure of RBC maturation
▪ Leads to megaloblastic anaemia in which the cells are larger and appear in megaloblastic (proerthroblastic) stage

HAEMATOCRIT VALUE
PROTEINS AND AMINO ACIDS: synthesis of protein part of haemoglobin
IRON: formation of heme part of haemoglobin
COPPER: Absorption of iron from GIT
Cobalt and Nickel: Utilization of iron
Vitamins C ; haemoglobin formation

HAEMOGLOBIN AND IRON SYNTHESIS

▪ Haemoglobin is a chromoprotein
▪ Content is highest at birth
▪ The affinity of haemoglobin for co2 is 20 times that of oxygen
▪ Oxygen binds with haemoglobin to form oxyhaemoglobin in a process called oxygenation

HAEMOGLOBIN STRUCTURE
▪ Made of protein and iron component
▪ Protein part is globin while iron containing part is heme

HEME
▪ Pigmented part of heme is pophyrin
▪ Formed by 4 pyrolle rings attached to one another by methane bridges
▪ Iron is attached to the nitrogen of all pyrolle rings

GLOBIN
▪ 2 alpha and beta chains in adults but two alpha and gamma chains in foetus
▪ Fetaal haemoglobin has more affinity for oxygen
▪ Oxygen dissociation curve for haemoglobin is shifted to the left

SYNTHENSIS OF HEME
▪ Heme is synthensized from succinyl COA and the glycine

FATE OF HAEMOGLOBIN
▪ After 120 days, the RBC is destroyed in the reticuloendothelial cells
▪ Particularly in spleen and haemoglobin its released into plasma
▪ Haemoglobin is degraded in the reticuloendothelial cells and split into globin and heme
▪ Globin is utilised again for the synthesis of haemoglobin
▪ Heme is degraded into iron and porphyrin
▪ Iron is stored as ferritin and hemosiderin in small quantites and are reutilised
▪ Porphyrin is converted into a green pigment called bilverdin and is converted into bilirubin
▪ Bilverdin and bilirubin are called bile pigments

ABNORMAL HAEMOGLOBIN
▪ Caused by abnormal changes in the polypeptide chain
▪ HAEMOGLOBINOPATHY: caused by abnormal polypeptide chains
▪ THALASSEMIA: Decrease in polypeptide chains, in alpha alpha chains decreased,absent and abnormal and vice versa for beta
▪ ABNORMAL HAEMOGLOBIN DERIVATIVES:
Carboxyhaemoglobin: Carbon monoxide plus hemoglobin and it results in tissue hypoxia

METHMOGLOBIN: Iron molecule of haemoglobin is oxidized from ferrous to ferric state, less than 40 percent of the body cels
▪ Body stands a chance of continual production but its hindered by erythrocyte protective system (Nicotinamide Adenine
Dinucleotide)
▪ Operates through 2 systems diaphorase 1 and diaphorase 2, due to dependency of NADH dependent reductase or presence of
abnormal haemoglobin M, blue baby syndrome may occur

SULFHEMOGLOBIN: Formed by combination of haemoglobin with iron sulphide and it cannot be reversed. When it gets to a
level of 10, it becomes toxic

IRON METABOLISM
▪ Major component of haemoglobin
▪ Total quantity of iron in the body is about 4g
▪ Dietary iron is available in heme and none heme
▪ Heme is easily absorbed while non heme is not easily absorbed
▪ Iron is absorbed mainly from the small intestine
▪ Bile is essentizal for absorption of iron
▪ Iron is present mainy in ferric form and its converted to ferrous form which is mainly absorbed by the blood
▪ Hcl from gastric juice makes the ion more soluble so that it can b converted to ferric ion by ferric reductase
▪ Ferric ion is transported into the blood by ferroportin
▪ After absorption in the blood, it combines with beta globulin called apoferritin resultiing in the transfer of transferrin
▪ Iron is transported in the blood in form of transferrin
▪ Iron is stored in the blood in form of ferritin in large amount and hemosiderin in small quantity

DAILY IRON LOSS

 ▪ 1mg excreted through faeces
▪ Iron loss is much in females during menstruation
▪ Each gram of haemoglobin contains 3.34g of iron

ERYTHROCYTE SEDIMENTATION RATE

This is the rate of settlement of red blood cell without being centrifuged.
It is also called sedimentation rate, sed rate or biernaci reactio

It depends on:
• Shape of the cells
• Concentration of plasma proteins
• Infection
The stacking of red blood cell on one another is termed Rouleux formation.

FACTORS THAT INCRASE ESR

• Specific gravity: specific gravity increases weight of RBC and so settling down is faster
• Roleaux Formation: piling up makes RBC heavier and it makes it settle down

FACTORS THAT DECRASE ESR

• Increased viscosity decreases ESR
• Increase in number of RBC decreases viscosity which inturn

USES
• Monitoring recovery from diseases or efficiency of treatment.
• Varies with ages and sex.
• Normal values are
• New born = 2mm/hr
• Adult male = 3-7mm/hr (5.7)
• Adult female = 3-15mm/hr (9.5)

NOTES ABOUT ESR

• Increased NUMBER OF RBC will lead to a drop in ESR
• Increased in the NUMBER OF RBC would lead to viscosity
• Less in children and infants due to increased RBC
• More in females because of less RBC
• Increases during menstruation because of less RBC
• Decreases in allergic conditions, polycythemia, severe leucytosis

PACKED CELL VOLUME

• True propotion of blood occupied by RBC expressed in percentage

PACKED CELL VOLUME DETERMINATION

• Blood is mixed with EDTA or heparin and filled in hematocrit
• Tube is centrifuged for 3000 per minute for 30 mins

VARIATIONS IN PACKED CELL VOLUME

INCREASES
• Increases in polycythemia, dehydration and shock syndrome
DECREASES
• Decreases in anaemia, pregnancy and haemorrhage

BLOOD INDICES
• They are calculations derived from RBC count

MEAN CORPSCULAR VOLUME

• This is the average volume of a single RBC expressed in cubic microns
• Normal MCV is 90 CU (78-90)
• Normal MCV makes RBC= Normocyte
• Increased MCV= Macrocyte
• Reduced MCV= Microcyte
• Pernicious and Megaloblastic Anaemia= Macrocytic and normochromic or hypochromic
• Iron deficiency Anemia= Microcytic and hypochromic
• Protein deficiency anemia= macrocytic and hypochromic

MEAN CORPSCULAR HAEMOGLOBIN

• Quantity of hemoglobin present in one RBC

MEAN CORPSCULAR HAEMOGLOBIN CONCENTRATION

• Concentration of hemoglobin present in one RBC
• Normal- Normochromic
• Decreases- Hypochromic
• Its not possible to be hyperchromic because the volume of RBC cannot increase beyond normal

COLOUR INDEX
• Ratio btw the number of RBC and hemoglobin
• Average hemoglobin content in cell of a patient as compared to that of a normal person
• Increases in pernicious and megaloblastic anemia
• Reduced in iron deficiency anemia
• Normal in normocytic, normochromic anemia

ANEMIA
• A blood disorder characterized by decrease in RBC,HEMOGLOBIN CONTENT AND PCV
• It leads to reduced supply of oxygen
• Oocurs as a result of decreased RBC production,Destrution of RBC and Excess blood loss

MORPHOLOGICAL ANEMIA
• Size is determined by mean corpuscular volume
• Colour is determined by mean corpuscular haemoglobin concentration

ETIOLOGICAL CLASSIFICATION
• HEMORRHAGIC ANEMIA: This is due to excessive loss of blood
• Occurs in acute and and chronic haemorrage
ACUTE ANEMIA: Occurs in sudden loss of blood
CHRONIC ANEMIA: Occufrs due to excessive loss of blood by internal or external bleeding and lot of blood is lost causing iron deficiency,
cells become small and RBC becomes microcytic and hypochromic
HEMOLYTIC ANEMIA: destrcution of RBC not compesated by increased RBC Production and its caused by extrinsic and intrinsic factors
ALPHA THALASSEMIA: ALPHA CHAIN are absent but Beta chains are excess in adults and GAMMA in children
BETA THALASSEMIA: Beta chains are less with alpha chain less in number

NUTRITIONAL DEFICIENCY ANEMIA

• IRON DEFICIENCY: Caused by ininadequate availability of iron for haemoglobin synthesis
• RBC are microcytic and hypochromic
• Difficulty in swalloeing= dysphagia and other features include brittle hairs,atrophy iof pappila in tongue
• PROTEIN DEFICIENCY: Haemoglobin synthesis is reduced, RBC is macrocytic and hypochromic
• PERNICIOS/ADDISON ANEMIA: Caused by the deficiency of maturation factor for RBC, It is due to the atrophy of gastric mucosa because
of parietal cells
• Gastric atrophy results in the decreased production of intrisic factors
• Pernmcicious anemia is associated with auto immune disorders
• APLASTIC ANEMIA: Disorder of bone marrow where RBC is reduced and replaced by fatty tissues. RBC are normocytic and normochroic
• ANEMIA OF CHRONIC DISEASE: 2ND common type of anemia charcterized by short lifespan of RBC
• Caused by distortion in iron metabolism and resistance to erythropetin factor and is cuased by tuberculosis.chronic renal failure and
neoplastic disorders

HEMOLYSIS AND FRAGILITY OF RBC

• It is the rupture of RBC with the release of hemoglobin into blood

HEMOLYSIS IN NORMAL CONDITION

• Rupture occurs after 120 days
• Cell membrane becomes fragile and it ruptures in the spleen
• Only 10% of RBC are hemolyzed this way

HEMOLYSINS
• They are substance which destroy RBC'S
• Gram positive and gram negative bacteria
• Phospholipase A2 present in the venom of poisonous snakes causes hemolysis by lysin phospholipid part of cell membrane
• Chemical substances such as benzene, NSAIM

HEMOLYSIS IN ABNORMAL CONDITION

• Younger RBC are reduced and broken down, this leads to hemolytic jaundice and hemolytic anemia
• Abnormal shape of RBC and auto immune diseases
• Mechanical factors such as physical stress and compression of muscles

FRAGILITY
• This is the susceptibility of RBC to hemolysis
• Osmosis fragility occurs due to disturbance in fragile medium

FRAGILITY TEST
• Used to measure RBC resistance in saline solution
• Done by using NACL solutions

WHITE BLOOD CELLS

• Colourless and nucleated formed elements of the blood
• They are responsible for body defence
• Leukocytes are divided into granulocytes and agranulocytes
 GRANULOCYTES
• They are the WBC having granules in their cytoplasm
• They are divided into neutrophils,eosinopjils and basophills
AGRANULOCYTES
• They are WBC having plain cytoplasm without granules

MORPHOLOGY OF RBC
NEUTROPHILS
• Small granules in the cytoplasm
• Granules take both acid and basic stain and appear violet in solution
• Nucleus is multillobed and number depends on cell age with no lobes in children but 2-5 in older ones
• Ameboid in nature
• Half life is 60 hours
• Diameter is 10-12
ESINOPHILS
• Diameter is 10-14
• Hlaf life is 10-20 hours
• Nucleus is bilobed
• They stain pink or reddish orange
BASOPHILS
• Diameter is 8-10
• Stain purple blue with methylene blue
• Nucleus is bilobed
MONOCYTES
• Largest leucocytes
• Diameter is 14-18
• Clear cytoplasm without granules
• Nucleus is pushed to one side
LYMPHOCYTES
• Nucleus occupies whole part of cytoplasm
• Cytoplasm may or may not be seen
• Divided into large(young) with a diameter of 10-12 and small (older)7-10
• They are divided into T and B lymphocytes
• T is concerned with cellular immunity and B with humoral immunity

NORMAL WBC COUNT

• Total WBC is 4,000-11,000 CC of blood

LEUKOCYTOSIS
• It is the Increase in WBC
• Occures in physiological and pathological conditions
• Occurs in conditions like infections, allergy, tuberculosis ,grandular fever

LEUKOPENIA
• Decrease in WBC Count
• Pathological conditions alone like anemia, anaphylatic shock, and disorders of spleen

PHYSIOLOGIAL VARIATIONS
• More in infants aand less in adults
• Slightly more in males and females
• Count is higher in the afternoon than in the morning
• Increases slightly during exercise, menstruation and paturation and anxiety
• Decreases during sleep

PROPERTIES OF WBC
• DIAPEDSIS: Process by which leucocytes squeeze through blood vessels
• They show ameboid movement characterized by potrusion of cytoplasm
• CHEMOTAXIA: This is the attraction of WBC towards injured sites by chemiacl substances released at the sites of injury
• PHAGOCYTES: Neutrophils and monocytes en.gulf by means of phagocytosis
•
FUNCTIONS OF NEUTROPHILS
• First line of defense along with monocytes
• They are free cells

MECHANISM OF ACTION OF NEUTROPHILS

• Neutrophils are released in large number at the site of infection from the blood.
• At the same time, new neutrophils are produced from the progenitor cells
• All the neutrophils move by diapedesis towards the site of infection due to chemotaxis aided by chemoattractants
• After reaching the area, the neutrophils surround the area and get adhered to the infected tissues.
• Chemoattractants increase the adhesive nature of neutrophils so that all the neutrophils become sticky and get attached firmly to
the infected area.
• Each neutrophil can hold about 15 to 20 microorganisms at a time.
• Now, the neutrophils start destroying the invaders.
• First, these cells engulf the bacteria and then destroy them by means of phagocytosis

RESPIRATORY BURST
• Respiratory burst is a rapid increase in oxygen consumption during the process of phagocytosis by neutrophils and other phagocytic
cells.
• Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase is responsible for this phenomenon.
• During respiratory burst, the free radical O2– is formed. 2O2– combine with 2H+ to form H2O2 (hydrogen peroxide).
• Both O2-– and H2O2 are the oxidants having potent bactericidal action.
Pus and Pus Cells
• Pus is the whitish yellow fluid formed in the infectedtissue by the dead WBCs, bacteria or foreign bodies and cellular debris.

ESINOPHILS
• defense mechanism of the body against the parasites.
• During parasitic infections, there is a production of a large number of eosinophils which move towards the tissues
• affected by parasites.
• Eosinophil count increases also during allergic diseases like asthma

BASOPHILS
• Basophils play an important role in healing processes.
• So their number increases during healing process.
• Basophils also play an important role in allergy or acute hypersensitivity reactions (allergy).
• This is because of the presence of receptors for IgE in basophil membrane.

Mechanism of Action of Basophils

Functions of basophils are executed by the release of some important substances from their granules such as:
1. Heparin: Heparin is essential to prevent the intravascular blood clotting.
2. Histamine, slow-reacting substances of anaphylaxis, bradykinin and serotonin: Theses substances produce the acute
hypersensitivity reactions by causing vascular and tissue responses.
3. Proteases and myeloperoxidase: These enzymes destroy the microorganisms.

MONOCYTES
• Largest cell among leucocytes
• First line of body defense
• Precursors of tissue macrophages
• Examples are kupfer cells, macrophages in lungs and spleen

• They are activated by lymphokines from T-lymphocytes and are called histocytes or wandering cells.

• they are much more powerful phagocytes than neutrophils.

• Macrophages have a powerful lysosomal lipase which breaks down the lipid-rich cell memebranes of many bacteria.

MAST CELL
• Large tissue cell resembling basophil
• Majorly found around blood vessels
• Their cells do not enter the blood stream
• Plays a role in hypersensitivity reactions
• When activated, mast cells release chemical mediators into interstituim from granules
• PERFOMED MEDIATORS: Already formed and stored in secretory granules
• Newly generated mediators: Arachidonic acid derivatives such as leukptriene C

LEUKOPOESIS
• Development and maturation of leukocytes
• Stimulated by hematopoetic growth factors and colony stimulating factors (CFS)
• GCSF= Granulocytes CSF secreted by monocytes and endothelial cells
• GMCSF= Granulocytes- Monocyte CSF secreted by monocytes, endothelial cells and T lymphocytes
• MCSF= Monocyte CSF secreted by monocytes and endothelial cells

IMMUNITY
• Immunity is the capacity of blood to resist pathogenic agent
• Immunity is divided into innate immunity and acquired immunity

INNATE IMMUNITY OR NON SPECIFIC IMMUNITY

• Inborn capacity of the body to resist pathogens
• It eliminates the development of any disease

ACQUIRED IMMUNITY OR SPECIFIC IMMUNITY

• This is the resistance in the body developed against certain disease

DEVLEOPMENT OF LYMPHOCYTES
• T lymphocytes or T cells are responsible for cellular immunity
• B lymphocytes or B cells are rresponsible for Humoral immunity
 T - LYMPHOCYTES
• It enters thymus and is processed into T lymphocytes
• Thymosin accelerates the proliferation and activates lymphocytes in thymus
PROCESSING
• Lymphocytes from bone marow enter cortex of thymus and they start proliferaton
• We have a rearranagement of polypeptide chains on the receptor protein called T Cell Receptors present on Lymphocyte surfaces
• T lymphocytes recognize the foreign antigen on surface of antigen presenting cells
• During rearrangement in majority we have 1 alpha and 1 beta chain or 1alpha and 1 gamma
BETA SELECTION
• Thymocytes having a rearranged beta chain undergo further processing
• The ones that fail beta selection undergo apoptosis
DOUBLE POSITIVE AND DOUBLE NEGATIVE
• Thymocytes with beta selection pass to medulla
• Cell start expressing another types of molecules such as CD4,CD8,CD25
• Cells that CD4 and CD8 are double positive
• CD25 and CD44 are double negative
• The ones
POSITIVE SELECTION AND NEGATIVE SELECTION
• CD4 and CD8 undergo certain processing are allowed to undergo further processing into T lymphocytes
• The ones that do not express CD 4 and CD 8 are eliminated by apoptosis
SINGLE POSITIVE CELLS
• Cells develop into single positive T lymphocytes
• Some become suppressor and some become helper

T - LYMPHOCYTES
Helper cells or inducer cells contain CD4+T cells
Cytotoxic or killer cells have CD8 T cells
Suppressor T cells
Memory T cells

STORAGE OF T LYMPHOCYTES
• They leave thymus and are storred in lymphoid tissues

B - LYMPHOCYTES
• Processed in liver during foetal life and bone marrow after birth
• Transformed into plasma and memory cells after processing

ANTIGENS
• They produce specific immune reactions
SELF ANTIGENS: They are antigens present on the body own cells such as A antigen and B antigen in RBC
NON SELF ANTIGENS: They enter the body from outside e.g
• Antigenic cells present on cell membrane of microbial organisms
• Toxins from microbial organisms
• Allergens like pollen grains

TYPES OF NON SELF ANTIGENS

• They induce development of immunity or production of antibodies- immunogenicity
• React with specific antibodies and produce allergic reactions- allergic reactivity

CHEMICAL NATURE OF ANTIGENS

• They are conjugated proteins like lipoproteins ,glycoproteins and nucleoproteins

DEVELOPMENT OF CELL MEDIATED IMMUNITY

• It is the immunity developed by cell mediated response
• Involves Tlymphocyes, macrophages and natural killer cells
• This kind of immunity does not involve antigens
• It is ofefered by Tlymphocytes and starts developing when Tcells come in cocntact witj antigens
• Invading materials carry antigenic materials and are released from invading organisms
• They are presented to cells by helper T cells
• Responsible for delayed allergic infections and rejection of transplanted tissues

ANTIGEN PRESENTING CELLS

• They are special type of cells in the body that stimulates the release of antigen presenting cells and they also present them to the T
helper cells
• They are macropgahes (phagocytic), dendritic cells (non phagocytic) and B-lymphocytes
• Dendritic cells includes cells of spleen that traps antigens in spleen, follicular dendritic cells in lymph ndes and langerhans dendritic
cells in skin
• B-LYMPHOCYTES: They rae both antigen presenting and antigen receiving and they need to be activated by helper T cell

ROLE OF ANTIGEN PRESENTING CELLS

• They are either englufed by phagocytosis or trapped by dendritic cells
• Antigen is digested into small peptide products
• This product moves towarD the surface of antigen presenting cells and bind with human leucocyte antigen (HLA)
• HLA is a genetic class found in moleccule of class II major histocompatibilty complex (MHC) situated on the surface of antigen
presenting cells

MHC and HLA

• MHC is a large molecule found in chromosome 6
• HLA is made up of of MHC
• CLASS 1 MHC MOLECULE is present in all nucleated cells of the body and is involved in the presentation of antigenic peptides to cytotoxic
T cells
• Class 2 MHC MOLECULE is found in B cells, macrophages and other antigen presenting cells. Involved in presentation of antigenic
peptides to helper T cell

PRESENTATION OF ANTIGEN
Antigen presentaing cell presents its class II MHC molecules together with antigen bound HLA to helper T cells
▪ This inturn activates T helper cells
Sequence of Events during Activation of Helper T cells
• Helper T cell recognizes the antigen displayed on the surface of the antigen-presenting cell with the help of its own surface
receptor protein called T cell receptor.
• Recognition of the antigen by the helper T cell initiates a complex interaction between the helper T cell receptor and the
antigen. This reaction activates helper T cells.
• At the same time, macrophages (the antigen-presenting cells) release interleukin-1, which facilitates the activation and
proliferation of helper T cells.
• Activated helper T cells proliferate and the proliferated cells enter the circulation for further actions
5. Simultaneously, the antigen which is bound to class II MHC molecules activates the B cells also, resulting in the development of
humoral immunity

ROLE OF HELPER T CELLS

• Helper T cells (CD4 cells) which enter the circulation and activate all the other T cells and B cells.
• Normal, CD4 count in healthy adults varies between 500 and 1500 per cubic millimeter of blood.
Helper T cells are of two types:
1. Helper-1 (TH1) cells
2. Helper-2 (TH2) cells.
Role of TH1 Cells
TH1 cells are concerned with cellular immunity and secrete two substances:
i. Interleukin-2, which activates the other T cells.
ii. Gamma interferon, which stimulates the phagocytic activity of cytotoxic cells, macrophages and natural killer (NK) cells

Role of TH2 Cells

TH2 cells are concerned with humoral immunity and secrete interleukin-4 and interleukin-5, which are concerned with:
i. Activation of B cells.
ii. Proliferation of plasma cells.
iii. Production of antibodies by plasma cell.

ROLE OF CYTOTOXIC T CELLS

• They are activated by T-HELPER cells and they circulate through blood,lymph and destroys invading organisms

MECHANISM OF ACTION OF THE CYTIO-TOXIC T CELLS

▪ Receptors located on the outer membrane of the cyto-toxic T cells binds with antigens
▪ The cytotoxic T cells enlarge and produce lysozomal enzymes
▪ The substances destroy the invading the invading organisms
OTHER ACTIONS OF CYTO-TOXIC T CELLS
▪ They destroy cancer cells and other foreign BODIES
▪ They are the killer T cells
▪ They destroy virus most especially as the antigen of the virus attracts the T cells

ROLE OF SUPPRESSOR T CELLS

• They suppress the T killer cells and they prevent them from damaging the body own tissues along with invaded organisms
• Suppressor Cells also suppress HELPER CELLS TOO

ROLE OF MEMORY T CELLS

• They do not enter circulation but remain in lymphoid tissues
• When the body is exposed to same organism, they quickly activate other T cells

SPECIFICITY OF T CELLS
• A Tcell can only be activated by a tyoe of antigen

DEVELOPMENT OF HUMORAL IMMUNITY

• Humoral immunity is mediated by antibodies secreted into blood and lymph
• Blood and fluids are known as body fluids
• Macrophages and other antigen presenting cells are involved in humoral immunity

PRESENTATION OF ANTIGEN
• Presents antigenic peptides bound with HLA to B cells
ACTIVATION OF B CELLS
• With the help of B cell receptor, it recognizes the antigen on the surface of the antigen presenting cell
• Complex interaction between B cell receptor and antigen activates B cells
• Macrophages releaese interlukin 1 which facilitates activation and proliferation of B cells
• Antigen bound to class II MHC activates helper T cells

TRANSFORMATION OF B CELLS
• They are transformed into plasma cells and memory cells

PLASMA CELLS
• They produce antibodies
• 2000 molecules per second
• They are released into the lymph and transported into circulation

ROLE OF MEMORY B CELLS

• Occupy lymphoid in inactive form
• Forms the basic principle of immunization
• During second exposure more quantities of antibodies are produced than the first time
ROLE OF HELPER T CELLS
• They are simultaneously activated by antigens
• They secrete interlukin-2 and B growth factor which promote:
ACTIVATION O F GREATER NUMBER OF B LYMPHOCYTES
PROLIFERATION OF PLASMA CELLS
PRODUCTION OF ANTIBODIES
ANTIBODIES OR IMMUNOGLOBULINS
• They are proteins produced by B lymphocytes in response to the presence of an antigen
• They form 20% of total plasma proteins
• They are gamma globulin in nature

TYPES OF ANTIBODIES
• IgG forms 75 percent of antibodies in the body

STRUCTURE OF ANTIBODIES
• They are made up of 2 chains
• One pair of heavy chain and one pair of light chains'
• The heavy chains are connected to each other by disulfide bonds
• The heavy chains are connected to the light chains by disulfide bonds
• Heavy chain has 420 amino acids while light chains has around 220 amino acids
• They have 2 regions; namely the constant and variable region

CONSTANT REGION
• Identification and functions of immunoglobulins depend on this region
• Also called the complement binding region
• Binds to antibody receptor on cell membrane surface
• Amino acids present in this region are similar in number and placement

VARIABLE REGION
• Amino acids occupying this region are different in number and sequence
• This region enables the antibody to recognise and bind with antigen
• This region is smaller
Functions of Different Antibodies
1. IgA plays a role in localized defense mechanism in external secretions like tear.
2. IgD is involved in recognition of the antigen by B lymphocytes.
3. IgE is involved in allergic reactions.
4. IgG is responsible for complement fixation.
5. IgM is also responsible for complement fixation.

MECHANISM OF ACTIONS OF ANTIBODIES

• Direct actions and complement systems

DIRECT ACTIONS
Agglutination: foreign surfaces with antigen are clumped together
Precipitation: Soluble antigens like tetanus toxins are converted into insoluble forms and precipitated
Neutralization: antibodies cover the toxic sites of antigenic products
Lysis Potent antibodies rupture the cell membrane of organisms and rupture them

COMPLEMENT SYSTEM:
• Indirect actions are stronger than than direct actions
• It is a system of plasma enzymes
• They are 11 in number and are activated in 3 ways: classical, lectin and alternate
CLASSICAL PATHWAY (OLCA)
• C1 binds with antibodies and triggers a series of events in which other enzymes are activated in sequence
• OPSONIZATION: Activation of neutrophils and macrophages to engulf the bacteria with the use of an enzyme OPSONIN
• LYSIS: Destruction of bacteria by rupturing the cell membrane
• AGGLUTINATION: This is the clumping of RB C and bacteria
• NEUTRALIZATION: Covering toxic sites of antigenic products

LECTIN PATHWAY
• Mannose binding lectin binds with fructose on the wall of bacteria

ALTERNATE PATHWAY
• Protein in circulation (factor 1) binds with the polysaccharides present in cell membrane of invading organisms
• This inturn activates C3 and C5 which attacks antigenic products of invading organisms

SPECIFICITY OF B lymphocyes
• A Bcell can only be activated by a tyoe of antigen

NATURAL KILLER CELLS

• Large granular cell involved in defemse mechanism
• It has an indented nuclues
• It is the third kind of lymphocytes
• Also known as the non t or b cells
• It is the first line of defense immunity against virus
• It is not a phagocytic cell but it contains hydrolytic enzymes
• This causes the lysis of invading organisms

CYTOKINES
• They are small proteins that act as cell signalling molecules
• These proteins are by WBC
• They activate and regulate the general immune system of the body

IMMUNIZATION
• This is simply preparing the body to fight against a disease
• Divided into active and passive immunization

PASSIVE IMMUNIZATION
• Produced without challenging the immune system of the body
• Done by adminstering serum and beta globulin

NATURAL PASSIVE IMMUNIZATION

• Acquired from mother after birth
• Before birth Ig G through placenta
• After birth Ig E through breastmilk
• Lymphocytes of the child not activated and antibodies received from mother are soon metablolized

PASSIVE ARTIFICAIL IMMUNIZATION

• Developed by using previously activated antibodies
• Serum containing antibody is administered to people who have developed the disease (Prophyxlasis)

ACTIVE IMMUNIZATION
• Immune system of body is activated
• Body develops resistance agianst disease

CLINICAL INFECTION
• A disease with signs and symptoms casued by invading pathogenic organisms

SUBCLINICAL INFECTION
• Symptoms are very mild and do not alert affected subject

VACCINES
• Induces immunity either by activation of T lymphocytes and production of antibodies

TOXIODS
• Processsed to destroy toxicity but retains its capacity to induce antibody production by immune system
• Consists of waekend toxins produced by pathogens

IMMUNE DEFICIENCY DISEASE

• Group of disease in which components of immune system are defective
• They are congenital immune system and acquired immune deficiency disease

CONGENITAL IMMUNE DEFICIENCY DISEASE

• They occur due to effect in B cell or T cell
• DI george syndrome occurs due to absence of thymus and severe combines immune deficiency (due to absence of lymphoid tissue)

ACQUIRED IMMUNE DEFICIENCY DISEASE

• Occurs when CD4+T cells count is below 200 cells per cubic millimeter
• Initiation occurs when a glycoprotein from HIV binds to the surface receptor of T lymphocytes and macrophages leading to
destruction of the cells
• Causes a reduction in immune function and gives chances to opputunistic diseases

AUTO IMMUNE DEFICIENCY DISEASE

• Immune system mistakenly destroys its own cells
• Tolerance is when a immune system fails to give response to an antigen
• When tolerance fails, auto immunity happens
• It leads to activation of T lymphocytes and production of antibodies from B lymphocytes
• Auto immune disease occurs when the normal body tolerance decreases and the immune system fails to recognise the body tissues as
self

HUMAN LEUCOCYTE ANTIGEN SYSTEM

• It is a group of antigens on chromosome 6
• They encode the proteins that transport antigens from within the cell towards cell surface
• HLA molecules are recognised by T and B lymphocytes
• Antibodies are directed against tissues processing HLA

COMMON AUTOIMMUNE DISEASES

• Insulin dependent diabetes miletus common in childhood
• Myasthenia gravis due to development of antibodies against receptors for acetychloine in neuromuscular junction
• Hashimoto thyroiditis commomn in middle aged women and it impairs the formation of thyroid follicles leading to hypothyroidism
• Graves disease occurs when autoantibodies activate TSH receptors leading to hyperthyroidism
• Rheumathoid arithritis is due to chronic inflamation of synovial lining of joints leading to swelling around joints and tendons
• Chronic inflammation occurs due to the continous production of autoantibodies called rheumatoid arthritis factors (RAF)

ALLERGY AND IMMUNOLOGICAL SENSITIVITY REACTION

• Allergy is an abnormal immune response to a chemical or a physical agent
• During first exposure, the body doesn’t produce any immune response because sensitization is required for the reaction
• Subsequent exposure to the antigen would cause a variety of inflammatory response
• Immunological sensitivity reaction can be innate or acquired but are mostly mediated by antibodies

ALLERGENS
• It is anything that produces the manifestation of an allergy

IMMUNOLOGICAL HYPERSENSITIVE REACTION

• This kind of reactions give rise to several allergic conditions and auto immune disease
▪ They are classified into five
• TYPE 1 or ANAPHYLACTIC REACTIONS
▪ ASEJU (Exagerrated reactions of a body to an antigen that the body is already exposed to already
▪ It develops few minutes within an exposure to an antigen
▪ They are mediated by IgE and other factors involved in inflammation
▪ Inflammation is a protective response of tissues to damage
• TYPE 2 or CYTOTOXIC REACTIONS
▪ Mainly involve IgG antibodies and sometimes IgM and IgA
▪ The affected cells are destroyed
▪ Diseases due to cytotoxic reaction are haemolytic diseases
▪ Examples are RH incomptability and Autoimmune haemolytic anemia
• TYPE 3 OR ANTIBODY MEDIATED REACTIONS
▪ IgG or IgM are produced in this type
▪ Antibody-antigen complexes are precipitated and deposited in localized areas like joints and arthritis
• TYPE 4 OR CELL-MEDIATED REACTIONS
▪ It is also called delayed or slow type of hypersensitivy
▪ It is found in allergic reaction due to the bacteria, viruses and fungi
• TYPE 5 OR STIMULATORY/ BLOCKING REACTION
▪ Seen in auto immmune disease
PLATELETS
• They are small, colorless and non nucleated and moderately refractile formed elements in the blood
• Inactivated platelets are without processes while activated platelets have filipodia or PROCESSES

STRUCTURE OF PLATELETS
• They have a cell membrane, microtubles and cytoplasm

CELL MEMBRANE
▪ It is 6nm thick
▪ Extensive invaginattion forms an open canalicular system through which platelet granules extrude themselves
▪ It contains lipids in form of phospholipids glycoproteins and cholesterol
GLYCOPROTEINS: They prevent adherence of platelets to normal endothelium and accelerate the adherence of platelets to collagen
and ruptured endothelium.
▪ Glycoproteins form receptors for ADP and Thrombin
PHOSPHOLIPIDS
 ▪ They accelerate clotting reactions
▪ They form precursors of prostaglandins and Thromboxane A2

MICROTUBULES
▪ They are made up of tubulin
▪ They form a ring around the cytoplasm below the cell membrane
▪ They help inactivated platelets to maintain the disk like shape
CYTOPLASM
▪ Contains cellular organelles
▪ Enzymes like ATP and Prostaglandin needed enzymes
▪ Hormonal substances like adrenaline, Histamine, 5-hydroxytryptamine
▪ Platelet granules like alpha and dense granules
▪ Other chemical substances like glycogen, blood group antigens and inorganic substances like calcium
▪ NORMAL COUNT IS 2,50,000 CU/MM of blood and it ranges between 2,00,000 and 4,00,000 MM of bvlood
NOTES
▪ They are less in infants (1,50,000-2,00,000) and reaches normal level at 3rd month
▪ Count increases in high altitude
▪ Platelets count increases after eating food
▪ Reduces during menstruation
PROPERTIES OF PLATELETS

ADHESIVENESS:
▪ During injury to a blood vessel, endothelium is damaged and subendothelial collagen is exposed
▪ Platelets come in contact with collagen and are activated and they stick
▪ Adhesion involves interaction between von wilebrand factor secreted by damaged endothelium and a receptor protein
(Glycoprotein IB)
▪ This protein is situated on the surface of platelet membrane
▪ Collagen, thrombin, ADP, Thromboxane A2 Calcium ions E.t.c acceletrate adhesiveness

AGGREGATION
▪ This is the grouping of platelets
▪ Adhesion is followed by activation of many platelets by substances released from dense granules
▪ Platelets change their shape with elongation of long filamentous pseudopodia which are called processes
▪ Filipodia helps the platelet to aggregate together
▪ Its accelerated by thrombin, ADP, Thromboxane A2
AGGLUTINATION
▪ Clumping together of platelets
FUNCTIONS OF PLATELETS
• BLOOD CLOTTING: They are responsible for formation of intrinsic Prothrombin activator
• CLOT RETRACTION: Platelets are entrapped in between fibrin threads, and cytoplasm of platelets contains contractile proteins like
qctin,myosin and throbosthenin responsible for clot retraction
• HEMOSTASIS: During injury to a blood vessel and with injured endothelium and exposed collagen, platelets adhere to collagen and
induce hemostasis
• RUPTURED BLOOD VESSSEL REPAIR: Platelet derived growth factor helps to repair ruptured blood vessels
• DEFENSE MECHANISM: Platelets encircle foreign body and destroy them

ACTIVATORS OF PLATELETS
Collagen, thrombin, ADP, Thromboxane A2, Calcium ions, convulxin (purified protein from snake venom)
P selectin: Cell adhrsion molelcule secreted from endothlilial cells

INHIBITORS OF PLATELETS
Nitric oxide, Clotting factors, prostacylcin and nucleotidase which breaks down ADP

DEVELOPMENT OF PLATELETS
• They are formed from bone marrow
• Pluripotent stem cells give rise to colony forming unit megakaryocyte
• This develops into megakaryocyte
• Megakaryocyte cytoplasm forms pseudopodium
• A portion of psuedopodium detaches to form plateletes
• Production is influenced by colony stimulating factors and thrombopoetin
• colony stimulating factors are secreted by moncytes and T-Lymphocytes
• Thrombopoetin is a glycoprotein like erythropoetin and is secreted by liver and kidneys

LIFESPAN OF PLATELETS
• Average lifespan is 10 days but it ranges between 8 and 11 days
• Splenomegaly (spleen enlargement) decreases platelet count while splenectomy (removal of spleen) increases platelet count

HEMOSTASIS
• This is the process that results in arrest or stoppage of bleeding from a blood vessel
STAGES OF HEMOSTASIS
• Injury initiates hemostasis
• During injury, endothelium is damaged and collagen is exposed
• Adherence to collagen is accelerated by von willebrand factor
• This factor acts as a bridge between a special glycoprotein present on platelet surface and collagen fibrils
• Activated platelets induce hemostasis which occurs in 3 major phases

STAGES OF HEMOSTASIS
(VASOCONSTRICTION)
• When the injury occurs, blood vessels constrict immediately and decreases loss of blood from damaged portion
• It is a local phenomenom
• Platelets activated during this injury secretes serotonin (5Ht) which causes consritiction of blood vessels

(PLATELET PLUG FORMATION)

• Platelets adhered to collagen of ruptured blood vessel secretes ADENOSINE DIPHOSPHATE (ADP) and thromboxane A2
• This substance attracts more and more platelets

COAGULATION OF BLOOD
• Fibrinogen is converted into fibrin, Fibrin threads get attached to loose platelet plug
• Ruptured part of blood vessel is blocked and futher blood loss is presented

DEFINITION
• This is the process by which blood loses its fluidity and becomes a jelly like mass few minutes after it is shed out or put in a container
FACTORS INVOLVED IN BLOOD CLOTTING
• Coagulation of blood occurs due to activation of certain factors called clotting factors
▪ Factor 1- Fibrinogen
▪ Factor 2- Prothrombin
▪ Factor 3- Thromboplastin
▪ Factor 4- Calcium
▪ Factor 5- Labile factor
▪ Factor 6- Not visible
▪ Factor 7- Stable
▪ Factor 8- Anti hemophillic
▪ Factor 9- Christmas
▪ Factor 10- Stuart Prower
▪ Factor 11- Plasma Thromboplastin
▪ Factor 12- Hageman
▪ Factor 13- Fibrin Stabilizing
SEQUENCE OF CLOTTING MECHANISM

ENZYME CASCADE THEORY

Most enzyme present in blood clotting are present in inactive forms called pro-enzymes
Pro-enzymes must be converted into enzymes before reaction can occur
It’s a series of cascading enzymes in which one reaction precedes the following reaction
Stages of Blood Clotting
• Prothrombin activator
• Conversion of prothrombin to thrombin
• Conversion of fibrinogen to fibrin

STAGE 1: FORMATION OF PROTHROMBIN ACTIVATOR

• Blood clotting commences with the formation of a substance called prothrombin activator, which converts prothrombin into
thrombin.
• Its formation is initiated by substances produced either within the blood or outside the blood.
Thus, formation of prothrombin activator occurs through two pathways:
1. Intrinsic pathway.
2. Extrinsic pathway.

Intrinsic Pathway for the Formation of Prothrombin Activator

• In this pathway, the formation of prothrombin activator is initiated by platelets, which are within the blood itself.
Sequence of events in intrinsic pathway
• During the injury, the blood vessel is ruptured. Endothelium is damaged and collagen beneath the endothelium is exposed.
• When factor XII (Hageman factor) comes in contact with collagen, it is converted into activated factor XII in the presence of
kallikrein and high molecular weight (HMW) kinogen.
• The activated factor XII converts factor XI into activated factor XI in the presence of HMW kinogen.
• The activated factor XI activates factor IX in the presence of factor IV (calcium).
• Activated factor IX activates factor X in the presence of factor VIII and calcium.
• Now the activated factor X reacts with platelet phospholipid and factor V to form prothrombin activator. This needs the
presence of calcium ions.
• Factor V is also activated by positive feedback effect of thrombin

Extrinsic Pathway for the Formation of Prothrombin Activator

In this pathway, the formation of prothrombin activator is initiated by the tissue thromboplastin, which is formed from the injured
tissues.
 Sequence of events in extrinsic pathway
1. Tissues that are damaged during injury release tissue thromboplastin (factor III). Thromboplastin contains proteins,
phospholipid and glycoprotein, which act as proteolytic enzymes.
2. Glycoprotein and phospholipid components of thromboplastin convert factor X into activated factor X,in the presence of
factor VII.
3. Activated factor X reacts with factor V and phospholipid component of tissue thromboplastin to form prothrombin
activator. This reaction requires the presence of calcium ions.
STAGE 2: CONVERSION OF PROTHROMBIN INTO THROMBIN
Blood clotting is all about thrombin formation. Once thrombin is formed, it definitely leads to clot formation.
Sequence of Events in Stage 2
1. Prothrombin activator that is formed in intrinsic and extrinsic pathways converts prothrombin into thrombin in the
presence of calcium (factor IV).
2. Once formed thrombin initiates the formation of more thrombin molecules. The initially formed thrombin activates Factor
V. Factor V in turn accelerates formation of both extrinsic and intrinsic prothrombin activator, which converts
prothrombin into thrombin. This effect of thrombin is called positive feedback effect
STAGE 3: CONVERSION OF FIBRINOGEN INTO FIBRIN
• Final stage of blood clotting involves the conversion of fibrinogen into fibrin by thrombin.
Sequence of Events in Stage 3
i. Thrombin converts inactive fibrinogen into activated fibrinogen due to loss of 2 pairs of polypeptides from each fibrinogen
molecule. The activated fibrinogen is called fibrin monomer.
ii. Fibrin monomer polymerizes with other monomer molecules and form loosely arranged strands of fibrin.
iii. Later these loose strands are modified into dense and tight fibrin threads by fibrin-stabilizing factor (factor XIII) in the presence
of calcium ions (Fig. 18.1). All the tight fibrin threads are aggregated to form a meshwork of stable clot.

BLOOD CLOT
DEFINITION AND COMPOSITION OF CLOT
• Blood clot is defined as the mass of coagulated blood which contains RBCs, WBCs and platelets entrapped in fibrin meshwork.
• The RBCs and WBCs are not necessary for clotting process.
• However, when clot is formed, these cells are trapped in it along with platelets.
• The trapped RBCs are responsible for the red color of the clot.
• External blood clot is also called scab.
• It adheres to the opening of damaged blood vessel and prevents blood loss.

CLOT RETRACTION
• Clot retraction is the process which involves contraction of blood clot 30 to 45 minutes after formation and oozing of a straw-colored
fluid called serum out of clot.
• Contractile protein, namely thrombosthenin in the cytoplasm of platelets is responsible for clot retraction.

FIBRINOLYSIS
• Fibrinolysis is the process that involves breakdown and dissolution of blood clot inside the blood vessel.
• It helps to remove the clot from lumen of the blood vessel.
• Fibrinolysis requires a substance called plasmin or fibrinolysin.

Formation of Plasmin
• Plasmin is formed from inactivated glycoprotein called plasminogen.
• Plasminogen is synthesized in liver and is incorporated with other proteins in the blood clot.
• Plasminogen is converted into plasmin by tissue plasminogen activator (t-PA), lysosomal enzymes and thrombin.
• The t-PA and lysosomal enzymes are released from damaged tissues and damaged endothelium. Thrombin is derived from blood. The
t-PA is always inhibited by a substance called t-PA inhibitor.
• It is also inhibited by factors V and VIII.
• Besides t-PA, there is another plasminogen activator called urokinase plasminogen activator (u- PA). It is derived from blood.

Sequence of Events Involved in Activation of Plasminogen

1. During intravascular clotting, the endothelium of the blood vessel secretes a thrombin-binding protein, the thrombomodulin.
It is secreted by the endothelium of all the blood vessels, except the minute vessels of brain.
2. Thrombomodulin combines with thrombin and forms a thrombomodulin-thrombin complex.
3. Thrombomodulin-thrombin complex activates protein C.
4. Activated protein C inactivates factor V and VIII in the presence of a cofactor called protein S.
5. Protein C also inactivates the t-PA inhibitor.
6. Now, the t-PA becomes active.
7. Activated t-PA and lysosomal enzymes activate plasminogen to form plasmin.
Plasminogen is also activated by thrombin and u-PA

ANTICLOTTING MECHANISM IN THE BODY

Under physiological conditions, blood does not clot during circulation because of the following reasons:
1. Smooth surface of endothelium of blood vessels prevents activation of clotting factors
2. Glycocalyx layer on inner surface of endothelium repels platelets and clotting factors and thereby initiation of blood clotting is
prevented
3. Continuous flow of blood does not allow aggregation of platelets and prevents blood clotting
4. Presence of natural anticoagulants in blood:
i. Heparin: It is the natural anticoagulant that is produced by the liver.
ii. Protein C: Thrombomodulin is produced by damaged endothelium of the blood vessels (except in brain capillaries). Thrombomodulin
is a thrombin-binding protein. It binds with thrombin and forms a thrombomodulin-thrombin complex, which activates protein C.
Activated protein C along with its cofactor protein S inactivates Factor V and Factor VIII.
• Inactivation of these two clotting factors prevents clot formation.
ANTICOAGULANTS
Anticoagulants are the substances which prevent or postpone clotting blood.
Anticoagulants are of three types:
1. Anticoagulants used to prevent blood clotting inside the body, i.e. in vivo.
2. Anticoagulants used to prevent clotting of blood that is collected from the body, i.e. in vitro.
 3. Anticoagulants used to prevent blood clotting both in vivo and in vitro.

1. HEPARIN
• Heparin is a naturally produced anticoagulant in the body.
• It is produced by mast cells which are the wandering cells present immediately outside the capillaries in many tissues or organs that
contain more connective tissue.
• They are abundant in liver and lungs.
• Basophils also secrete heparin.
• Heparin is a conjugated polysaccharide.
• Commercial heparin is prepared from the liver and other organs of animals.
• Commercial preparation is available in liquid form or dry form as sodium, calcium, ammonium or lithium salts.

Mechanism of Action of Heparin

Heparin
i. Prevents blood clotting by its antithrombin activity. It directly suppresses the activity of thrombin.
ii. Combines with antithrombin III (a protease inhibitor present in circulation) and removes thrombin from circulation.
iii. Activates antithrombin III.
iv. Inactivates the active form of other clotting factors like IX, X, XI and XII (Fig. 18.3).

Uses of Heparin
Heparin is used as an anticoagulant both in vivo and in vitro.

Clinical uses
Intravenous injection of heparin (0.5 to 1 mg/kg body weight) postpones clotting for 3 to 4 hours (until it
is destroyed by the enzyme heparinase). So, it is widely used as an anticoagulant in clinical practice.

Heparin is used for many purposes such as:

i. To prevent intravascular blood clotting during surgery.
ii. While passing the blood through artificial kidney for dialysis.
iii. During cardiac surgery involving heart-lung machine.
iv. To preserve the blood before transfusion.

Laboratory Use
Heparin is also used as anticoagulant in vitro while collecting blood for various investigations.
About 0.1 to 0.2 mg is sufficient for 1 mL of blood. It is effective for 8 to 12 hours.
After that, blood will clot because heparin only delays clotting and does not prevent it

2. COUMARIN DERIVATIVES
Warfarin and dicoumarol are the derivatives of coumarin.

Mechanism of Action
Coumarin derivatives prevent blood clotting by inhibiting the action of vitamin K. Vitamin K is essential
for the formation of various clotting factors, namely II, VII, IX and X.

Uses
Dicoumarol and warfarin are the commonly used oral anticoagulants (in vivo).
Warfarin is used to prevent myocardial infarction (heart attack), strokes and thrombosis.

3. EDTA
Ethylenediaminetetraacetic acid (EDTA) is a strong anticoagulant. It is available in two forms:
i. Disodium salt (Na2 EDTA).
ii. Tripotassium salt (K3 EDTA).
Mechanism of ActionT
These substances prevent blood clotting by removing calcium from blood.
Uses
EDTA is used as an anticoagulant both in vivo and in vitro. It is:
i. Commonly administered intravenously, in cases of lead poisoning.
ii. Used as an anticoagulant in the laboratory (in vitro). 0.5 to 2.0 mg of EDTA per milliliter of blood is sufficient to preserve the blood
for at least 6 hours. On refrigeration, it can preserve the blood up to 24 hours

4. OXALATE COMPOUNDS
• Oxalate compounds prevent coagulation by forming calcium oxalate, which is precipitated later.
• Thus, these compounds reduce the blood calcium level
• mixture of ammonium oxalate and potassium oxalate in the ratio of 3:2 is used.
• Each salt is an anticoagulant by itself.
• But potassium oxalate alone causes shrinkage of RBCs AND Ammonium oxalate alone causes swelling of RBCs.
• But together, these substances do not alter the cellular activity
Mechanism of Action
Oxalate combines with calcium and forms insoluble calcium oxalate.
Thus, oxalate removes calcium from blood and lack of calcium prevents coagulation.
Uses
Oxalate compounds are used only as in vitro anticoagulants. 2 mg of mixture is necessary for 1 mL of blood.
Since oxalate is poisonous, it cannot be used in vivo.

5. CITRATES
Sodium, ammonium and potassium citrates are used as anticoagulants.
Mechanism of Action
Citrate combines with calcium in blood to form insoluble calcium citrate.
citrate also removes calcium from blood and lack of calcium prevents coagulation.

Uses
Citrate is used as in vitro anticoagulant:
i. It is used to store blood in the blood bank as:
a. Acid citrate dextrose (ACD): 1 part of ACD with 4 parts of blood.
b. Citrate phosphate dextrose (CPD): 1 part of CPD with 4 parts of blood.
Citrate is also used in laboratory in the form of formol-citrate solution (Dacie’s solution) for RBC and platelet counts.

6. OTHER SUBSTANCES WHICH PREVENT BLOOD CLOTTING

Peptone, C-type lectin (proteins from venom of viper snake) and hirudin (from the leech Hirudinaria manillensis) are the known
anticoagulants.

PHYSICAL METHODS TO PREVENT BLOOD CLOTTING

Coagulation of blood is postponed or prevented by the following physical methods.
1. COLD: Reducing the temperature to about 5°C postpones the coagulation of blood.
2. COLLECTING BLOOD IN A CONTAINER WITH SMOOTH SURFACE: Collecting the blood in a container with smooth surface like a silicon-
coated container prevents clotting. The smooth surface inhibits the activation of factor XII and platelets. So, the formation of
prothrombin activator is prevented.

PROCOAGULANTS
Procoagulants or hemostatic agents are the substances which accelerate the process of blood coagulation.
Some procoagulants are given below:
1. THROMBIN
Thrombin is sprayed upon the bleeding surface to arrest bleeding by hastening blood clotting.
2. SNAKE VENOM
Venom of some snakes (vipers, cobras and rattle snakes) contains proteolytic enzymes which enhance blood clotting by activating
the clotting factors.
3. EXTRACTS OF LUNGS AND THYMUS
Extract obtained from the lungs and thymus has thromboplastin, which causes rapid blood coagulation.
4. SODIUM OR CALCIUM ALGINATE
Sodium or calcium alginate substances enhance blood clotting process by activating the Hageman factor.
5. OXIDIZED CELLULOSE
Oxidized cellulose causes clotting of blood by activating the Hageman factor.

TESTS FOR BLOOD CLOTTING

Blood clotting tests are used to diagnose blood disorders. Some tests are also used to monitor the patients treated with anticoagulant
drugs such as heparin and warfarin:
1. Bleeding time.
2. Clotting time.
3. Prothrombin time.
4. Partial prothrombin time.
5. International normalized ratio.
6. Thrombin time.

APPLIED PHYSIOLOGY

THROMBOSIS
Thrombosis or intravascular blood clotting refers to coagulation of blood inside the blood vessels.
Normally, blood does not clot in the blood vessel because of some factors which are already explained. But
some abnormal conditions cause thrombosis.
Causes of Thrombosis
1. Injury to blood vessels
During infection or mechanical obstruction, the endothelial lining of the blood vessel is damaged and it initiates thrombosis.
2. Roughened endothelial lining
In infection, damage or arteriosclerosis, the endothelium becomes rough and initiates clotting.
3. Sluggishness of blood flow
Decreased rate of blood flow causes aggregation of platelets and formation of thrombus.
Slowness of blood flow occurs in reduced cardiac action, hypotension, low-metabolic rate, prolonged confinement to bed and
immobility of limbs.
4. Agglutination of RBCs
Agglutination of the RBCs leads to thrombosis. Agglutination of RBCs occurs by the foreign antigens or toxic substances.
5. Toxic thrombosis
Thrombosis is common due to the action of chemical poisons like arsenic compounds, mercury, poisonous mushrooms and snake
venom.
6. Congenital absence of protein C
Protein C is a circulating anticoagulant, which inactivates factors V and VIII. Thrombosis occurs in the absence of this protein.
Congenital absence of protein C causes thrombosis and death in infancy.

Complications of Thrombosis
1. Thrombus
During thrombosis, lumen of blood vessels is occluded. The solid mass of platelets, red cells and/or clot, which obstructs the blood
vessel, is called thrombus. Thrombus formed due to agglutination of RBC is called agglutinative thrombus.
2. Embolism and embolus
Embolism is the process in which the thrombus or a part of it is detached and carried in bloodstream and occludes the small blood
vessels, resulting in arrests of blood flow to any organ or region of the body. Embolus is the thrombus or part of it, which arrests the
blood flow. The obstruction of blood flow by embolism is common in lungs (pulmonary embolism), brain (cerebral embolism) or heart
(coronary embolism).
3. Ischemia
Insufficient blood supply to an organ or area of the body by the obstruction of blood vessels is called ischemia. Ischemia results in
tissue damage because of hypoxia (lack of oxygen). Ischemia also causes discomfort, pain and tissue death. Death of body tissue is
called necrosis.
4. Necrosis and infarction
Necrosis is a general term that refers to tissue death caused by loss of blood supply, injury, infection, inflammation, physical agents or
chemical substances. Infarction means the tissue death due to loss of blood supply. Loss of blood supply is usually caused by occlusion of
an artery by thrombus or embolus and sometimes by atherosclerosis

BLEEDING DISORDERS
Bleeding disorders are the conditions characterized by prolonged bleeding time or clotting time.
Bleeding disorders are of three types:
1. Hemophilia.
2. Purpura.
3. von Willebrand disease
1. Hemophilia
• Hemophilia is a group of sex-linked inherited blood disorders, characterized by prolonged clotting time.
• However, the bleeding time is normal.
• Usually, it affects the males, with the females being the carriers.
• Because of prolonged clotting time, even a mild trauma causes excess bleeding which can lead to death.
Causes of hemophilia
Hemophilia occurs due to lack of formation of prothrombin activator. That is why the coagulation time is prolonged.
The formation of prothrombin activator is affected due to the deficiency of factors VIII, IX or XI.

Types of hemophilia
Depending upon the deficiency of the factor involved, hemophilia is classified into three types:
1. Hemophilia A or classic hemophilia: Due to the deficiency of factor VIII. 85% of people with hemophilia are affected by
hemophilia A.
2. Hemophilia B or Christmas disease: Due to the deficiency of factor IX. 15% of people with hemophilia are affected by
hemophilia B.
3. Hemophilia C or factor XI deficiency: Due to the deficiency of factor XI. It is a very rare bleeding disorder.

Symptoms of hemophilia i. Spontaneous bleeding. ii. Prolonged bleeding due to cuts, tooth extraction and surgery. iii. Hemorrhage in
gastrointestinal and urinary tracts. iv. Bleeding in joints followed by swelling and pain. v. Appearance of blood in urine.
Treatment for hemophilia
Effective therapy for classical hemophilia involves replacement of missing clotting factor.

2. Purpura
• Purpura is a disorder characterized by prolonged bleeding time. However, the clotting time is normal.
• Characteristic feature of this disease is spontaneous bleeding under the skin from ruptured capillaries.
• It causes small tiny hemorrhagic spots in many areas of the body.
• The hemorrhagic spots under the skin are called purpuric spots (purple colored patch-like appearance).
• Blood also sometimes collects in large areas beneath the skin which are called ecchymoses.
Types and causes of purpura
Purpura is classified into three types depending upon the causes:
1. Thrombocytopenic purpura
Thrombocytopenic purpura is due to the deficiency of platelets (thrombocytopenia). In bone marrow disease, platelet production is
affected leading to the deficiency of platelets.
2. Idiopathic thrombocytopenic purpura
Purpura due to some unknown cause is called idiopathic thrombocytopenic purpura. It is believed that platelet count decreases due to
the development of antibodies against platelets, which occurs after blood transfusion.
iii. Thrombasthenic purpura
Thrombasthenic purpura is due to structural or functional abnormality of platelets. However, the platelet count is normal. It is
characterized by normal clotting time, normal or prolonged bleeding time, but defective clot retraction

3. von Willebrand Disease

• von Willebrand disease is a bleeding disorder, characterized by excess bleeding even with a mild injury.
• It is due to deficiency of von Willebrand factor, which is a protein secreted by endothelium of damaged blood vessels and
platelets.
• This protein is responsible for adherence of platelets to endothelium of blood vessels during hemostasis after an injury.
• It is also responsible for the survival and maintenance of factor VIII in plasma.
• Deficiency of von Willebrand factor suppresses platelet adhesion.
• It also causes deficiency of factor VIII.
• This results in excess bleeding, which resembles the bleeding that occurs during platelet dysfunction or hemophilia.

BLOOD GROUPS
INTRODUCTION
When blood from two individuals is mixed, sometimes agglutination (clumping) of RBCs occurs.
Agglutination is because of the immunological reactions.
the discovery of blood groups WAS by the Austrian Scientist Karl Landsteiner, in 1901.
Determination of ABO blood groups depends upon the immunological reaction between antigen and
antibody.
LANDSTEINER’S LAW
Landsteiner’s law states that:
1. If a particular agglutinogen (antigen) is present in the RBCs of a person, corresponding agglutinin (antibody) must be absent in the
serum.
2. If a particular agglutinogen is absent in the RBCs, the corresponding agglutinin must be present in the serum.
Though the second part of Landsteiner’s law is a fact, it is not applicable to Rh factor.

You do not have antibodies that react with the antigens of your own RBCs, but you do have antibodies for any antigens that your
RBCs lack.
For example, if your blood type is B, you have B antigens on your red blood cells, and you have anti-A antibodies in your blood plasma.
The agglutinogens are mucopolysaccarides, which are not only present on the red blood cells, but also, in body secretions, such as,
saliva, gastric juice and in tissues of liver, kidney and lungs.

ABO SYSTEM
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.
• Blood having antigen A belongs to ‘A’ group. This blood has β-antibody in the serum.
• Blood with antigen B and α-antibody belongs to ‘B’ group.
 • If both the antigens are present, blood group is called ‘AB’ group and serum of this group does not contain any antibody.
• If both antigens are absent, the blood group is called ‘O’ group and both α and β antibodies are present in the serum.

IMPORTANCE OF ABO GROUPS IN BLOOD TRANSFUSION

• During blood transfusion, only compatible blood must be used.
• The one who gives blood is called ‘donor’ and the one who receives the blood is called ‘recipient’.
• While transfusing the blood, antigen of the donor and the antibody of the recipient are considered.
• The antibody of the donor and antigen of the recipient are ignored mostly.
• Thus, RBC of ‘O’ group has no antigen and so agglutination does not occur with any other group of blood. So, ‘O’ group blood can
be given to any blood group persons and the people with this blood group are called ‘universal donors’.
• Plasma of AB group blood has no antibody. This does not cause agglutination of RBC from any other group of blood.
• People with AB group can receive blood from any blood group persons. So, people with this blood group are called ‘universal
recipients’.

INHERITANCE OF ABO AGGLUTINOGENS AND AGGLUTININS

• Blood group of a person depends upon the two genes inherited from each parent.
• Gene A and gene B are dominant by themselves and gene O is recessive.
• Agglutinogens appear during the 6th month of fetal life.
• Concentration at birth is 1/5 of the adult concentration.
• It rises to the adult level at puberty.
• Agglutinins are the gamma globulins which are mainly IgG and IgM immunoglobulins.

Rh FACTOR
Rh factor is an antigen present in RBC. This antigen was discovered by Landsteiner and Wiener.
It was first discovered in Rhesus monkey
Rh group system is different from ABO group system because, the antigen D does not have corresponding natural antibody (anti-D).
However, 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.

INHERITANCE OF Rh ANTIGEN
Rhesus factor is an inherited dominant factor.
It may be homozygous Rhesus positive with DD or heterozygous Rhesus positive with Dd (Fig. 19.2).
Rhesus negative occurs only with complete absence of D (i.e. with homozygous dd).

APPLIED PHYSIOLOGY

TRANSFUSION REACTIONS DUE TO ABO INCOMPATIBILITY

Transfusion reactions are the adverse reactions in the body transfusion of incompatible (mismatched) blood.
The reactions may be mild causing only fever and hives (skin disorder characterized by itching) or may be severe leading to renal
failure, shock and death.
In mismatched transfusion, the transfusion reactions occur between donor’s RBC and recipient’s plasma.
So, if the donor’s plasma contains agglutinins against recipient’s RBC, agglutination does not occur because these antibodies are
diluted in the recipient’s blood.
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.

Severity of Transfusion Reactions

• Severity of transfusion reactions varies from mild (fever and chills) to severe (acute kidney failure, shock and death).
• Severity depends upon the amount of blood transfused, type of reaction and general health of the patient.

Cause for Transfusion Reactions

• Transfusion of incompatible blood produces hemolytic reactions.
• The recipient’s antibodies (IgG or IgM) adhere to the donor RBCs, which are agglutinated and destroyed.
• Large amount of free hemoglobin is liberated into plasma.
• This leads to transfusion reactions.

Signs and Symptoms of Transfusion Reactions

Non-hemolytic transfusion reaction
Non-hemolytic transfusion reaction develops within a few minutes to hours after the commencement of blood transfusion.
Common symptoms are fever, difficulty in breathing and itching.
Hemolytic transfusion reaction
Hemolytic transfusion reaction may be acute or delayed. The acute hemolytic reaction occurs within few minutes of transfusion. It
develops because of rapid hemolysis of donor’s RBCs. Symptoms include fever, chills, increased heart rate, low blood pressure, shortness
of breath, bronchospasm, nausea, vomiting, red
urine, chest pain, back pain and rigor. Some patients may develop pulmonary edema and congestive cardiac failure.
Delayed hemolytic reaction occurs from 1 to 5 days after transfusion.
The hemolysis of RBCs results in release of large amount of hemoglobin into the plasma.