Human Anatomy & Physiology

WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
ABO BLOOD GROUP SYSTEM lysis or destruction of the red blood cells of the
donor in the patient’s body
TOPIC OUTLINE
1 Introduction Why there will be lysis or destruction of the red
2 History blood cells?
3 Landsteiner’s Law
4 ABO Blood Group Antigens  This is due to the antibodies present in the
5 ABO Antibodies patient or recepient because antigens present
6 Routine ABO Testing from the cell of the donor will be recognized as
7 Inheritance of ABO Blood Groups foreign to the patient. One must take note that
the patient already has the antibodies against
8 Blood Group Mythology
these cells. So the patient’s antibodies will
9 ABO Discrepancies
attack the red blood cells of the donor to be
10 Technical Errors
transfused because of the incompatibility. This
will cause the lysis of the donor’s cells since the
wrong blood type was transfused to the
ABO BLOOD GROUP SYSTEM patient
 There are different other blood group systems, but  This produces a very severe, or even fatal,
the ABO system is said to be the most important of all transfusion reaction to the patient
blood groups in both transfusion and transplant
medicine  And even today, transfusion of the wrong ABO blood
group remains the leading cause of death in hemolytic
 This is the only blood group system in which transfusion reaction fatalities
individuals already have antibodies in their serum to
antigens that are absent from their red blood cells
HISTORY OF THE ABO BLOOD GROUP SYSTEM
ANTIGENS
 Substances that are recognized by the body as HISTORY
something foreign which can cause an immune
response.

 This could be bacteria, viruses or anything that a

body will recognize as foreign. However, these blood
group antigens are usually found on the membrane
of the red blood cells of individuals

ANTIBODIES
 Substances produced by the body in response to
the antigen or the foreign substance that was
detected by the body. We usually develop antibodies
when we get exposed to the antigen

 This occurs without any prior exposure to red blood

cells through transfusion or pregnancy
 When the person will be transfused with blood,
he/she will already be exposed to the different
antigens or antibodies present in the blood of
the donor or where blood came from
 Due to the presence of these antibodies, transfusion
of an incompatible ABO blood type may result in
immediate lysis of donor red blood cells
 Karl Landsteiner discovered the ABO blood group
system in the 1900s
 He classified an individual’s erythrocytes into 4
principal types: Blood Type A, B, AB, and O
 This marked the beginning of the concept of
individual uniqueness defined by the RBC antigens
present on the RBC membrane

 Incompatible – Wrong blood type was

transfused to the patient. This will result in the

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ

LANDSTEINER’S LAW ILLUSTRATION

THE LAW STATES THAT:

If an agglutinogen (antigen) is present on the
red blood cell membrane, the corresponding
1
agglutinin (antibody) must be absent in the
plasma or serum

If an agglutinogen is absent on the red blood cell

2 membrane, the corresponding agglutinin must
be present in the plasma or serum

 In other words, if the individual has the antigen, that

individual will not have that corresponding antibody  A single red blood cell has about 2 million blood
group antigens on its cell membrane. But for this
 Example, if an individual has antigen A, he/she illustration, we will take the two antigens for example
will not have the anti-A antibody

 The interaction between the antigen and the

antibody results in the agglutination or clumping of the
red blood cells.

 Antigen is also called agglutinogen; antibody is also

called agglutinin

ABO BLOOD GROUP ANTIGENS

 The blood groups are named for the antigens

present or absent on the surface of the red blood cells
 Example: group A individuals have the “A”
antigen; the group B individuals have the “B”
antigen
 Presence or absence of these antigens depends on
an individual’s DNA
 All blood type individuals, expressed a precursor
oligosaccharide on the red blood cells
 In the illustration, the yellow and green symbols
attached to the red blood cell served as the
precursor oligosaccharides

 DNA express different types of enzymes

 These enzymes, now that are expressed by the DNA,

are known as transferases. From the name itself, its
function is to catalyze a series of reactions in which they
transfer sugar units

 Catalyze – It will hasten or speed up the

reaction
 All humans also expressed a transferase enzyme
 These enzymes are responsible for the type of
that transfers and attaches fucose to this precursor
antigen expressed on the RBCs
oligosaccharides

 Fucose is demonstrated as the blue symbol in the

illustration

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ

 Then, fucose will be attached to it through a

certain transferase enzyme. This forms your H
antigen.

 Then, there is another transferase enzyme that

will attach the N-acetylgalactosamine which now
forms the A antigen

 So if this A antigen is present in an individual,

then that person is considered to be blood type A

 Binding a fucose to this precursor oligosaccharide

results in the formation of the H antigen

Now let’s try to see the difference between the

antigens present in the different blood types

BLOOD TYPE B
 There has to be the presence of the H antigen
which is made up of precursor oligosaccharide
and fucose

 In blood group B individuals, a different enzyme

will attach which is the galactose, which is the
 As discussed earlier, the presence or absence of
sugar unit of the H antigen. And with the
these antigens depends on the DNA of an individual.
attachment of the galactose, antigen B is formed
Then these DNA expressed different types of
enzymes. Specifically, they are transferases in which
their function is to transfer sugar units

BLOOD TYPE A
 For blood type A individuals, their DNA
expressed a certain transferase enzyme, which
attaches N-acetylgalactosamine (NAGA) to the H
antigen

 The N-acetylgalactosamine is actually the

sugar unit. So with the attachment of N-
acetylgalactosamine, the carbohydrate chain
now becomes antigen A

 Therefore, a blood type A individual has the A BLOOD TYPE AB

antigen  In individuals with AB blood type or blood
group, both enzymes (that is, the one that
transfers N-acetylgalactosamine and the other
TO MAKE IT CLEAR that transfers galactose) are expressed
 First, there has to be the presence of the
precursor oligosaccharide.  Therefore, the red blood cells of these
individuals expressed both A and B antigens

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
 A paragloboside or glycan is the same basic
precursor material from which A, B, and H antigens all
originate

 Specific enzyme transferases elicited by an inherited

gene attach sugars to the paragloboside or glycan

 The H antigen is the precursor structure on which A

and B antigens are made

 H antigen acts as the acceptor molecule for the two

sugars that make the A and B antigens
BLOOD TYPE O
 Individuals having O blood group only have H  In other words, the H antigen is the building
antigens on the red blood cells block for both the A and B antigens

 These individuals do not have enzymes that  As shown in the illustration a while ago, there
transfers N-acetylgalactosamine and galactose has to be a formation of the H antigen first so
to the H antigens that the specific sugars can be attached to it

 Therefore, neither A nor B antigens are  Therefore, A and B antigens can’t be formed
expressed by the red blood cells of blood O group without the H antigen
individuals
 The H and Se genes are not part of the ABO system
 There is no such thing as O antigen. Only the
absence of both A and B antigens  Yet, their inheritance influenced the
expression of the A and B antigens

H gene must be inherited to form ABO antigens

1
on the RBCs

 Develop early in fetal life Se gene must be inherited to form ABO antigens
2
in secretions
 Developed in utero (uterus) at 5 to 6 weeks of
gestation during pregnancy
THE SPECIFICITY OF THE A AND B ANTIGEN IS
 Take note that these antigens are already DEFINED BY THE IMMUNODOMINANT SUGAR
detectable at this time PRESENT

 Expression of A and B antigens on the RBCs is fully BLOOD

H antigen + N-acetylgalactosamine
developed by 2 to 4 years of age and remains constant TYPE A
throughout life
BLOOD
H antigen + D-galactose
TYPE B
ABH ANTIGENS
BLOOD H antigen + N-acetylgalactosamine +
 Formation of ABH antigens results from the TYPE AB D-galactose
interaction of genes at three separate loci (ABO, H, and
Se)  H antigen only (no additional sugars
attached)
 These genes do not actually code for the production BLOOD
of antigens but rather produce specific TYPE O
 O blood group has the highest
glycosyltransferases that add sugars to a basic concentration of H antigen
precursor substance

 The sugars referred to here are the N-  Blood group O individuals inherit at least one H gene
acetylgalactosamine and the galactose and two O genes

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ

 The H gene elicits the production of an enzyme called ILLUSTRATION

⍺-2- L-fucosyltransferase that transfers the sugar L-
fucose to an oligosaccharide chain

 L-fucose is the sugar responsible for H specificity

 Antigens, similar to ABO blood group antigens like

 The genes do not produce the antigen itself, but antigen A or antigen B, are found in nature. They are
produces these glycosytransferase enzymes expressed by common microorganisms such as
those found in human intestines and in many food
H GENE stuffs. So we are exposed to these antigens early in
 Produces ⍺-2-L-fucosyltransferase that attaches the development of our immune system
the immunodominant sugar L-fucose to the
paragloboside or glycan and this forms the H  So let’s say these are bacterial cells having an
antigen antigen A and antigen B similar to human blood
group antigens on their surface
A GENE
 Codes for the production of ⍺-3-N-
acetylgalactosaminyltransferase which transfers
the N-acetyl—D-galactosamine sugar to the H
antigen, which then forms antigen A

B GENE
 Codes for the production of ⍺-3-D-
galactosyltransferase and attaches the D-
galactose sugar to the H antigen, which then forms
antigen B

 For example, an individual with blood type A. This

 ABH antigens are integral parts of the membranes of
RBCs, endothelial cells, platelets, lymphocytes, and
epithelial cells
 ABH-soluble antigens can also be found in all body
secretions
means that the red blood cells of this individual
expressed the A antigen on its cell membrane. This
individual is exposed to similar antigen A and also
antigen B of the microbial surface or a similar
bacterial antigen A and antigen B

 Remember that these ABH antigens are found

in the membranes of the red blood celss

ABO ANTIBODIES

 Antibodies are present or found in the serum or

plasma of the person

 Antibodies are formed when our immune system is

exposed to non-self antigens
 Since antigen A is already present and is
recognized by the body as self-antigen, the similar A
 Antibodies are present against the antigen which is
antigen on the microbial surface will also be
absent or missing on the cell membrane of the RBCs

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
recognized as self. Therefore, the body will not
produce anti-A antibodies
 However, these microbes also have antigen B and
remember that type A individuals do not express
antigen B. So that makes the antigen B absent or
missing on the red blood cells
 That’s why it is very important to be extra
careful during the processing or the testing of
the blood and even during the transfusion
process itself
 The blood bag must be double-checked or
even triple-checked if it is really the one
intended for the patient or recepient

ABO ANTIBODIES
BLOOD TYPE A INDIVIDUAL Anti-B

BLOOD TYPE B INDIVIDUAL Anti-A

BLOOD TYPE O INDIVIDUAL Anti-A and Anti-B

BLOOD TYPE AB INDIVIDUAL None

 So these microbial antigen B will be recognized as

non-self. Therefore, anti-B antibodies are produced
against these bacterial antigen B. Thus, an individual
with blood type A, anti-B antibodies are found in
their serum

 The same concept applies to the other blood types

 Individuals normally produce antibodies directed

against the A or B antigen absent from their RBCs

 These antibodies have been described as naturally

occurring because they are produced without any
exposure to RBCs

 ABO antibody production is initiated at birth, but

titers are generally too low for detection until infants
are 3 to 6 months old

 The results of ABO testing before 3 to 6 months

of age cannot be considered valid because
some or all the antibodies present maybe
maternal antibodies or the antibodies of the
mother that has cross the placenta and not
from the baby himself/herself
 Antibody production peaks between 5 and 10 years
of age and declines later in life
 Elderly people usually have lower levels of
anti-A and anti-B
 There is still a reaction when blood type O is mixed
with the other blood types, however it is just minor or
weak and is referred to as clinically significant
 This is because of the type of antibodies of the ABO.
They are IgM antibodies and usually do not produce
very severe reactions
 The type of antibodies that can produce severe
transfusion reactions are the IgG antibodies found in
other blood group systems

 ABO antibodies can cause rapid intravascular  Blood type O is usually used for emergency purposes
hemolysis if the wrong ABO group is transfused, to the event the patient really needs to be transfused
potentially resulting in patient death with blood

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
 Blood type AB can receive blood from all the blood
types because they do not have both anti-A and anti-B TO EASILY REMEMBER
that can react with the antigens present in the other COLOR OF REAGENT ANTIBODY
blood types BLUE (anti) ANGEL
YELLOW (anti) BIRD
 Blood type O can only receive blood from type O
because it has both anti-A and Anti-B which can also  The samples and the reagent are mixed on a slide
react with the antigens from the other blood types with an applicator stick or a wooden stick and then
tilted back and forth & observed over a period of 2
RECAP minutes for the agglutination or clumping of the cells
ABO ANTIGENS ABO ANTIBODIES
Detectable at 5-6 weeks Detectable at 3-6  Longer periods of incubation should be avoided
of gestation months old because the effects of drying may be interpreted as
agglutination (pseudoagglutination or false
Fully developed by 2-4 Production peaks at 5- agglutination)
years of age 10 years of age
 Since we will only be using a small sample
Remain constant
Decline later in life on the slide, it could esily dry out. Once it
throughout life
does, it may look like there is agglutination
Found on the Found in the or clumping even though there’s really
membrane of the RBCs serum/plasma none. That might lead to us reporting the
wrong blood type. Therefore, it is important
to observe the result right away or over a
period of 2 minutes only
ROUTINE ABO TESTING

 Is done to determine the blood type of a person

 This is a very critical step in the process of blood

transfusion

 There are two methods namely forward typing and

reverse typing

 Karl Landsteiner was the first individual to perform

both the forward and reverse typing

FORWARD TYPING
 Also known as Cell Grouping

 Uses known sources of commercial antisera (anti-

A, anti-B) to detect antigens on an individual’s RBCs

 Can be done on a slide or in a test tube at room

temperature

 Done by mixing unknown RBC with known typing

sera  There will be agglutination or clumping if both the
antibody and the corresponding antigen is present
 Use of commercial reagents: Anti-A (blue) & Anti-
B (yellow)  In the forward typing, if you can see agglutination
with a corresponding antibody or antisera, then that
is the blood type of the patient. If there is no
agglutination, then the blood type is O

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
REVERSE TYPING
 Also known as Serum Testing

 Defined as detecting ABO antibodies in the

patient’s serum by using known reagent RBCs,
namely A and B cells

 Patient’s serum is tested with suspensions of

known group A & B cells

 Usually, these cells are taken from known

blood type A and blood type B individuals.
These can be drawn from you or from the RECAP
MedTechs who are known to be blood type FORWARD TYPING REVERSE TYPING
A or blood type B Aka Cell Grouping Aka Serum Testing

 Done to crosscheck the results of forward typing Detects antigens Detects antibodies

 ABO blood grouping is the most frequently Sample: patient’s RBCs Sample: patient’s serum
performed test in blood bank
Reagents: anti-A & anti- Reagents: known A & B
 This is a very critical step in blood B typing sera cells
transfusion because transfusion of an
incompatible blood can cause hemolytic
transfusion reaction or can even be fatal INHERITANCE OF THE ABO BLOOD GROUPS

 Both ABO forward and reverse typing tests must
be performed on all donors and recipients
 There is always an inverse reciprocal
relationship between the forward and reverse type;
thus, one serves as a check on the other
The theory for the inheritance of the ABO blood
groups was first described in 1924 by indicating that an
individual inherits one ABO gene from each parent
and that these two genes determine which ABO
antigens are present on the RBC membrane
 The inheritance of ABO genes follows simple
Mendelian genetics

 ABO like most other blood groups systems is

codominant in expression

 An individual inherits one gene from each

parent. It can either be A, B, or O gene

 A and B genes are both dominant genes while

the O gene is considered a recessive gene

ILLUSTRATION

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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
 Let’s take for example, a father who is blood type
AB and a mother who is heterozygous for blood type
A
 Heterozygous – The two genes are not the
same. Like in the case of the mother, she
has the A and O genes
 Homozygous - If the two genes are the
same
 With this, their offsprings could possibly be:
 Blood type A (homozygous)
 Blood type AB
 Blood type A (heterozygous)
 Blood type B (heterozygous)
 In codominance, the dominant gene will be
expressed. Since A and B genes are the dominant
genes, the O gene will be mast in the presence of
these genes. In the presence of these A and B genes,
both dominant genes will be expressed as seen in
type AB individuals
 So another example, let’s have a father who is
blood type A which is heterozygous, and the mother
is blood type B which is also heterozygous.
 Their offsprings could be:
 Blood type AB
 Blood type B (heterozygous)
 Blood type A (heterozygous)
 Blood type O (homozygous)
 In blood type O individuals, they have to inherit
both O genes because with the presence of either A
or B gene, the dominant gene will always be
expressed
 Take note that:
 Blood type AB is always heterozygous
 Blood type O is always homozygous
 Both blood types A and B may either be
homozygous or heterozygous
 One position, or locus, on each chromosome 9 is
occupied by an A, B, or O gene
 The O gene is considered an amorph
 As no detectable antigen is produced in
response to the inheritance of this gene
 The group O phenotype is an autosomal recessive
trait with the inheritance of two nonfunctional O genes
QUICK GUIDE
GENOTYPE
 An individual’s actual genetic make-up
 We inherit one gene from each parent. That
is why it is always two letters. This is the
actual genetic make-up of an individual
 Example: AA, BO, OO, AB
 Two genes are the same = homozygous
 Two genes are different = heterozygous
PHENOTYPE
 The outward expression of genes
 Mostly composed of serologically
demonstrable antigens
 Example: blood type A or blood type B
 An individual who has the phenotype A can have the
genotype AA or AO while an individual with phenotype
B can have the genotype BB or BO
 Serologically, it is not possible to determine the
specific genotype from the phenotype of a blood type A
or B individual
 Family studies or molecular assays would need to be
performed to determine the exact genotype
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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
BLOOD GROUP MYTHOLOGY
 Associatations between ABH antigens and
practically any disorders known to man can be found
throughout medical literature
Have more pronounced to
BLOOD GROUP A
“hangover”
There is a high possibility of
BLOOD GROUP B
“criminality”
BLOOD GROUP O Are said to have “good teeth”
 There are also several papers, correlating blood
groups with personality traits
ABO DISCREPANCIES
 Occur when unexpected reactions are obtained in
the forward and/or reverse typing
 Can be due to problems with the patient’s RBCs or
patient’s serum, or problems with both the serum and
cells
 These unexpected reactions may be due to an
extrapositive reaction or a weak/missing
reaction in either the forward or reverse typing
 All ABO discrepancies must be resolved prior to
reporting a patient or donor ABO blood group
 To avoid hemolytic transfusion reactions
 Due to weakly reacting or missing antibodies as seen
in the cases of:
NEWBORNS
 ABO antibody production is not detectable until 3
to 6 months of age
 Only forward typing should be done since
antigens can already be detectable at 5-6
weeks of gestation while the baby is still
inside the mother’s womb
ELDERLY PATIENTS
 Production of ABO antibodies is depressed
 ABO atibodies declined later in life
TECHNICAL ERRORS
 Can also cause ABO discrepancies
EXAMPLES OF TECHNICAL ERRORS
Incorrect or inadequate identification of blood
 specimens, test tubes, or slides
Blood sample and test tube labeling errors

Failure to add reagents or failure to add sample

Contaminated reagents

 Addition of incorrect reagents or sample
Clerical errors or incorrect recording of results

 Failure to follow manufacturer’s instructions
RESOLUTION
 Serum and antiserum should always be
added first, followed by the patient reagent
RBCs to avoid:
 Reagent contamination
 Potential omission of either patient
sample or reagent
 So that we cannot forget to add the sample,
especially that the serum is colorless
 If you add the known red cells first, you might
get confused if you have already added the
patient’s serum or not
 Results must be recorded immediately after
obtaining them to avoid transcription errors
 Always examine reagent vials concurrently
while performing ABO testing and quality
control testing for possible contamination
 Make sure any and all technical factors that
may have given rise to the ABO discrepancy are
reviewed and corrected
 Obtain adequate information regarding the
patient’s age, diagnosis, transfusion history,
medications, and history of pregnancy
 If the discrepancy persists and appears to be
due to an error in specimen collection or
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Human Anatomy & Physiology
WEEK NUMBER 8 / VIDEO LECTURE (LECTURER: MS. MARIAN G TECSON.) / TRANSCRIBED BY: NIÑA DE LA CRUZ
identification, a new sample must be drawn
from the patient and all RBC and serum testing
must be repeated

 When a discrepancy is encountered,

all results must be recorded, but the
interpretation of the ABO blood type
must be delayed up until the
discrepancy is reolved

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