IMMUNOHEMATOLOGY

( BLOOD BANKING )
Definition of terms:
Review
A. Absorption
= removal of an unwanted
antibody from a serum; often used
interchangeably with adsorption.

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Adsorption
= providing an Ab with its
corresponding antigen under
optimal conditions so that the Ab
will attach to the Ag. It is thus
take out of the serum . Often used
interchangeably with absorption.
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Agglutination
= the clumping together of RBC or
any particulate matter resulting
from interaction of Ab & its
corresponding Ag.

Agglutinin = an Ab that agglutinates
cell.
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Agglutinogen
= a substance that stimulates the
production of an agglutinin, thereby,
acting as an Ag.
Antibody = a protein subs. developed in
response to, & interacting
specifically w/ an Ag. In Blood
banking it is found serum, from
either a commercial manufacturer or
a patient. It is secreted by plasma
cells.
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Apheresis
= a method of blood collection in
w/c whole blood is withdrawn, a
desired component separated and
retained, & the remainder of the
blood returned to the donor.
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Avidity
= is a measure of a functional
affinity of an antiserum for the
whole antigen.
Elution
= a process whereby cells that are
coated with Ab are treated in such
a manner as to disrupt the bonds
bet. the Ag and Ab.
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Antigen
= a substance that is recognized by
the body as being foreign thus it
can elicit immune response.
Fibrinolysin/ Plasmin
= has the ability to dissolve fibrin.
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Fibrinolysis
= dissolution of fibrin by
fibrinolysin caused by the action
of a proteolytic enzyme system
that is continually active in the
body but that is increased greatly
by various stress stimuli.
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Hemolysin
= an antibody that activates
complement leading to cell lysis.

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Hemagglutination
= agglutination of RBC or clumping
of RBC.
Hemaaglutinin = a protein in blood
serum that causes clumping of
RBC; also present in the surface
projection of some viruses.
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Hemolysis
= disruption of the red cell
membrane and the subsequent
release of hemoglobin into the
suspending medium or plasma.
Immunogen
= any substance capable of
stimulating an immune response.
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Immunogenecity

= a descriptive term indicating
the ability of an antigen to
stimulate an Ab response.

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Immunoglobulin
= one of a family of closely related
though not identical proteins that
are capable of acting as
antibodies. They are IgA, IgG, IgM,
IgD, IgE
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IgG
= is the main Ig in human serum
IgA
= is the principal Ig in exocrine
secretions such as saliva & tears.
IgM
= is a globulin formed in almost
every immune response during the
early period of the reaction.
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IgD
= may play a role in Ag recognition
& the initiation of Ab synthesis.
IgE
= is produced by the cell lining the
intestinal & respiratory tract & is
important in forming reagin.
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LECTIN
= proteins that are present in
plants ( usually seeds) w/c binds
specially to carbohydrates
determinants & agglutinate RBC
through their cell surface
oligosaccharide determinants.
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Phlebotomy
= to take blood from a person.

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Sensitization
= a condition of being made
sensitive to a specific substance(
e.g. an Ag) after the initial
exposure to the substance.
Transfusion = the injection of blood,
a blood component, saline, or
other fluids into the bloodstream.
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B. Allele
= one of the two or more different
genes that may occupy a specific
locus on a chromosome.
Chromosomes
= the structure w/n the nucleus
that contain a linear thread of
DNA, w/c transmits genetic
information.
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Chromosome
= Genes are arranged along the
strand of DNA and constitute
portions of DNA.
Codominant Gene
= a pair of genes in w/c neither is
dominant over the other, that
they are both expressed.
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Dominant gene
= a trait or characteristic that will
be expressed in the offspring even
though it is only carried on one of
the homologous chromosomes.

Gene= a unit of inheritance w/n a
chromosome.
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Genotype
= an individuals actual genetic
make- up.
Heterozygous
= processing different alleles at a
given locus.



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Homozygous
= possessing a pair of identical
alleles.
Locus
= the site of a gene on a
chromosome.
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Meiosis
= type of cell division of germ cells
in w/c 2 successive divisions of the
nucleus produce cell that contain
half the no. of chromosomes
present in somatic cells.
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Mitosis
= type of cell division in w/c each
daughter cell contains the same
number of chromosomes as the
parent cell. All cell except sex
cells undergo mitosis.
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Interphase
= resting stage from cell division.
Prophase
= coiling of DNA protein complex;
chromosome duplication.
Metaphase
= alignment of chromosome in
equatorial plane of cell.
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Telophase
= constriction develops at the
times of equatorial plane of
mother cells.
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Phenotype
= the outward expression of genes
(e.g a blood type). On blood cells,
serologically demonstrate Ags
constitute by the phenotype
except those sugar sites that are
determinant by transferases.
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Recessive gene
= a gene that, in the presence of
its dominant allele, does not
express itself. Expression of
recessive genes occurs when they
are inherited in the homozygous
genes or state.
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Zygote
= the single fertilized cells formed
by the union of two gametes.
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PHASES OF HEMOSTATIC
MECHANISM
Vasoconstriction
= is the initial response
(neurogenic) to vessel damage.
Platelet aggregation
= by its ability to clump & forms a
temporary plug to the blood flow
outlet.
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Coagulation
= by the activation of plasma
factors forming a permanent plug
w/c promotes vessel healing.
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DNA AND RNA
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DNA
= is composed of a pentose sugar
lacking in oxygen known as
Deoxyribose, nitrogenous bases
(guanine, cytosine, thymine &
adenine) & a phosphate fastened
together forming a double helix
structure.
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RNA
= is likewise composed of a
pentose sugar, ribose, nitrogenous
bases ( guanine, cytosine, thymine
replace by uracil & adenine) &
forms a single coiled structure.
= it is also contains phosphate
bonds or groups.
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Types or forms of RNA:
1. rRNA = 80%
2. tRNA = 15% - adaptor in protein
synthesis; identifying/ picking up
of codon.
3. mRNA = 5% - template of
CHON synthesis.

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Processes:
1. Replication/ duplication
= model: Watson- Crick
a. semi- conservative
b. conservative
Major enzymes:
a. DNA polymerase
b. DNA ligase
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2. Transcription
steps:
a. binding of RNA polymerase
( major enzyme for transcription)
to DNA.
b. initiation of polymerization
c. chain elongation
d. termination of synthesis
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3. Translation ( Protein synthesis
stage)
1. hydrogen bond between
complementary.
2. covalent bond between sugar
and phosphate
3. interaction between sugar and
phosphate.
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4. Interaction between DNA and
histones.
5. Van der Waals of hydrophobic
forces

.stabilizing factor of RNA
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Differences:
DNA RNA
Double stranded single stranded
Bigger smaller
Deoxyribose (sugar) ribose (sugar)
Tymine- adenine uracil- adenine
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PROTEIN SYNTHESIS
INITIATION
=involves a collection of processes
w/c starts in the nucleotides
sequence AGGAGGU; initiation
ends in the attachment of 1
st
tRNA
carrying amino acid.
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ELONGATION
= starts in the attachment of a 2
nd

tRNA w/ amino acid to a codon of
mRNA through the ribosomal
subunits; the amino acid binds w/
the first one & this processes will
continue until a suitable or the
dictated polypepetide chain is
reached.
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TERMINATION
= after repetition of the above
process, elongation will stop at
the last tRNA attached by the
mRNA as dictated by the
transcripted strand.
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IMMUNOHEMATOLOGY

= refers to immunologic reactions involving
red blood cells and blood components.
= immunologic properties and reactions of
all blood components and con
constituents.

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APPLICATION
1. Safe blood transfusion and organ
transplantation.
2. Understanding the pathogenesis,
diagnosis and prevention of Rh
immunization associated with
pregnancy.
3. Resolution of disputed parentage
problem.
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BLOOD BANKING

= embraces with the collection,
processing, preparation and
distribution of blood components
and its derivatives.
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Review on BLOOD
BLOOD
= nutritive fluid that circulates all
throughout the body.
= produce and mature in the bone
marrow, released into the
bloodstream where they play
important functions.
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An adult human has about 46 liters of blood
circulating in the body. Among other things,
blood transports oxygen to various parts of
the body.
Blood consists of several types of cells
floating around in a fluid called plasma.

The red blood cells contain hemoglobin, a
protein that binds oxygen. Red blood cells
transport oxygen to, and remove carbon
dioxide from, the body tissues.


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The platelets help the blood to clot, if you
get a wound for example.

The plasma contains salts and various kinds
of proteins.





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Functions:
1. O2 transport and facilitates CO2
excretion.
2. Transport of hormone from the
organ of production.
3. Excretion of waste product from
cellular metabolism.
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4. Regulation of body
temperature.
5. Regulation of acid- base balance.
6. Involved body defense
mechanism.
7. Blood coagulation and
hemostasis.
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Composition
A. Formed elements= 45% of total
or cellular elements volume

1. RBC 2. WBC
3. Platelets
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B. Fluid portion
= 55% of total volume
PLASMA OR SERUM
1. H20 = 91.5%
2. electrolytes
3. proteins
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Characteristics or Properties
1. Fluid form but coagulates in
vitro
2. Red in color due to Hb.
3. Thick and viscous (3.5-4.5u)
4. Slightly alkaline
( 7.35- 7.45 )
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5. SG
= 1.055 (1.045- 1.065)

6. Comprises 7.8% of total body
weight or 75-85ml/kg body weight
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RBC membrane
= blood group antigens found.
= consist of bilipid layer formed by
orderly arrangement of
phospholipids.
= embedded in the lipid bilayers are
protein and other complex
molecules such as carbohydrates.
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Proteins are:
1. Integral CHON = outside
(glycophorin)
2. Peripheral CHON= cytoplasmic
(e.g. spectrin)

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Biochemical Composition of Cell
Membrane
a. 52% CHON = for enzymatic
reaction
= structural stability
b. 89% CHO= cell specifity
c. 40% lipids
= selective permeability
= rigidity
= fluidity
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Factors affecting red cell
survival
1. Normal chemical composition.
2. Structural arrangement of
components.
3. Interaction of erythrocyte
membrane.
4. Deformability ( fluidity)

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4. Deformability
= due to decrease ATP level,
increase calcium deposition in cell
membrane (increase rigidity)
5. Permeability
= electrolyte and water balance.
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Metabolic pathways that takes
place in RBC
1. Anaerobic glycolysis
= generates 90% ATP
2. Pentose phosphate pathway
= generates 10% ATP
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2. PPP
Deficient PPP reduce glutathione
(insufficient to neutralize
intracellular oxidants)

result in globin precipitation
as aggregate ( Heinz bodies)

cell membrane damage & cell
destruction
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3. Methemoglobin reductase
pathway
a. NADH methemoglobin reductase*
b. NADPH methemoglobin reductase
Absence of methemoglobin
reductase= accumulation of
methemoglobin (Abnormal Hgb)
due to conversion of Fe++ in Hb
to Fe+++ w/c alters O2 transport
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4. Luebering- Rapoport Shunt
Pathway

= produce 2,3- diphosphoglycerate
( 2,3- DPG)
Increase 2,3- DPG level= decrease
Hb-O2 affinity w/c is good in
oxygen transport.
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Assignment:
1. What is gene mutation?
2. What are the types of gene
mutation? Explain
comprehensively, give example
if necessary.


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HISTORICAL BACKGROUD
1492 - Pope Innocent VIII, in Rome,
had an apoplectic stroke; became
weak and went into a coma. His
physician advised a Blood transfusion
as a therapeutic measure for the
Pope's illness. Employing crude
methods, the Pope did not benefit and
died by the end of that year.

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1615 - Andreas Libavius described
his technique of Blood transfusion.
It was unfortunately not adequately
publicized.



1665 - The first Blood transfusions of record
take place. Animal experiments conducted
by Richard Lower, an Oxford physician
started as dog-to-dog experiments and
proceeded to animal-to-human over the
next two years. Dogs were kept alive by
the transfusion of Blood from other dogs.
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1667 - Jean-Baptiste Denis in
France
= successful transfusions from sheep
to humans.
1678= Transfusion from animals to
humans, having been tried in many
different ways, was deemed to be
unsuccessful, and was subsequently
outlawed by the Paris Society of
Physicians because of reactions, many
resulting in death.



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1818 - James Blundell, a British
obstetrician, performed the first
successful transfusion of human
Blood to a patient for the
treatment of postpartum
hemorrhage. Using the patient's
husband as a donor, he extracted a
small amount of Blood from the
husband's arm and, using a syringe,
he successfully transfused the
wife.
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1840 - In London England, Samuel
Armstrong Lane, aided by
consultant Dr. Blundell, performed
the first successful whole Blood
transfusion to treat hemophilia.
1867 - English surgeon Joseph
Lister utilized antiseptics to control
infection during Blood transfusions.

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Fr. Of BB
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.1901 - Karl Landsteiner, an
Austrian physician, and the most
important individual in the field of
Blood transfusion, documented the
first three human Blood groups
(based on substances present on
the red Blood cells), A, B and O.
= Father of Immunohematology.

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1902 - A fourth main Blood type, AB
was found by A. Decastrello and A.
Sturli.

1907 - Hektoen suggested that the
safety of transfusion might be
improved by cross-matching Blood
between donors and patients to
exclude incompatible mixtures.

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Reuben Ottenberg = performed the
first Blood transfusion using
Blood typing and cross-matching.
= also observed the 'Mendelian
inheritance' of Blood groups and
recognized the universal utility of
group O donors.
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1908 Epstein & Ottenberg
= how Ags produced
= blood groups are inherited.
1909 Bateson
= blood groups are acquired.
1914 Hustin
= Sodium Citrate as
anticoagulant.




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1915 Liwisohn
= determined the minimum
amount of citrate to be used.
1916 Rous &Turner
= Citrate Dextrose
1934 Berstein
= supported the postulate of
Ottenberg & Epstein.
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1939- 40 Landsteiner Weiner
Livine & Stetson
= discovered Rh and HDN
1943 Loutit & Turner
= introduced ACD
1948-49 Morgan, Watkins & Kabat
= discovered ABO structure.
1957 Gibson
= introduced CPD.


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Immunohematology

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