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Lab 6
LAB TITLE: BLOOD TYPE SIMULATION AND ANALYSIS AND BLOOD
GROUP GENETICS PROBLEMS

Part 1: Blood Type Simulation and Analysis

INTRODUCTION:
Around 1900, Karl Landsteiner discovered that there are at least four different
kinds of human blood based on the presence or absence of specific antigens on the
surface of red blood cells (erythrocytes). These antigens have been designated as A
and B. Antibodies against antigens A and B begin to build up in the blood plasma
shortly after birth. The antibody levels peak at about eight to ten years of age, and
the antibodies remain present in declining amounts throughout the rest of life. A
person normally produces antibodies against those antigens that are not on his/her
erythrocytes but does not produce antibodies against those that are present on
his/her erythrocytes. Thus, a person with antigen A has anti-B antibodies; a person
with B antigens has anti-A antibodies; a person with neither antigen A or B (blood
type O) has both anti-A and anti-B antibodies; and a person with both antigens A
and B has neither anti-A nor anti-B antibodies. The individual’s blood type is
based on the antigens, not the antibodies, he/she has.

The four blood groups are known as types A, B, AB and O. Blood type O,
characterized by the absence of A or B antigens, is the most common in the United
States (45% of the population). Type A is next in frequency, found in 39% of the
population. The incidences of types B and AB are 12% and 4% respectively.
TABLE 1, ABO SYSTEM
Blood Type Antigens on Antibodies in Can Give Can Receive
Erythrocytes Plasma Blood to Blood from
(Red Blood Groups Groups
Cells)
A A Anti-B A, AB O, A
B B Anti-A B, AB O, B
AB A and B Neither Anti- AB O, A, B, AB
A nor Anti-B
O Neither A nor Both Anti-A O, A, B, AB O
B and Anti-B
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Process of Agglutination
Blood typing is performed with reagents, called antiserum, which contain high
levels of anti-A and anti-B antibodies. The simple test is performed as follows:
Several drops of each kind of antiserum are added to separate samples of blood. If
clumping occurs only in the suspension to which the anti-A serum was added, the
blood type is A. If clumping occurs only in the anti-B mixture, the blood type is B.
Clumping in both samples indicates that the blood type is AB. The absence of
clumping indicates that the blood is Type O
TABLE 2
CLUMPING REACTIONS OF ABO BLOOD-TYPING SERA

Reaction Reaction
Blood Type
Anti-A Serum Anti-B Serum
Clumping (+) No Clumping (-) Type A
No Clumping (-) Clumping (+) Type B
Clumping (+) Clumping (+) Type AB
No Clumping (-) No Clumping (-) Type O

Importance of Blood Typing

Early attempts to transfuse blood from one person to another produced varied
results. If incompatible blood types are mixed, erythrocyte destruction, clumping
and other problems can occur. For instance, if a person with Type B blood is
transfused with blood Type A, the recipient’s anti-A antibodies will attack the
incoming Type A erythrocytes. The Type A erythrocytes will be destroyed, and
hemoglobin will be released into the plasma. in addition, incoming anti-B
antibodies of the Type A blood may also attack the Type B erythrocytes of the
recipient with similar results. This problem may not be serious, unless a large
amount of blood is transfused.

The ABO blood groups and other inherited antigenic characteristics of red blood
cells are often used in medico-legal situations involving identification or disputed
paternity. In paternity cases a comparison of the blood groups of mother, child and
alleged father may exclude the man as a possible parent of the child. For example,
a child of blood type AB whose mother is type A could not have as a father a man
whose blood group is O. Blood typing does not prove that an individual is the
father of a child; it merely indicates whether or not he is a possible parent.
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Rh System
In the period between 1900 and 1940 a great deal of research was done to
discover the presence of other antigens in human red blood cells. In 1940,
Landsteiner and Wiener reported that rabbit sera containing antibodies for the red
blood cells of the Rhesus monkey would clump the red blood cells of most
humans. This antigen in humans, designated as the Rh factor with due respect to
the Rhesus monkey, was later found to exist as six antigens, which were given the
letters C, c, D, d, E, and e by Fischer and Race.
Of these six antigens, the D factor is responsible for the Rh+ condition. An
individual who possesses these antigens is designated Rh+; an individual who
lacks them is designated Rh-. The anti-Rh antibodies of the system are not
normally present in the plasma, but anti-Rh antibodies can be produced upon
exposure and sensitization to Rh antigens. There are several ways sensitization can
occur – for example, if Rh+ blood is transfused into an Rh- recipient, or when an
Rh- mother carries a fetus who is Rh+. In the latter case, some of the fetal Rh
antigens may enter the mother’s circulation and sensitize her so that she begins to
produce anti-Rh antibodies against the fetal antigens. In most cases sensitization to
the Rh antigens takes place toward the end of pregnancy, but because it takes some
time to build up high levels of anti-Rh antibodies the first Rh+ child carried by a
previously unsensitized mother is usually unaffected. However, if an Rh- mother or
previously sensitized mother (by a blood transfusion or by a previous Rh+
pregnancy) later carries an Rh+ fetus, maternal anti-Rh antibodies may enter the
fetal circulation, causing the agglutination and hemolysis of fetal erythrocytes.
This would result in a condition known as erythroblastosis fetalis (hemolytic
disease of the newborn). To treat an infant in a severe case, the infant’s Rh+ blood
is removed and replaced with Rh- blood from an unsensitized donor. The
replacement of the infant’s blood reduces the levels of the anti-Rh antibodies.
The genetics of the Rh blood group system is complicated by the fact that
more than one antigen can be identified as the result of the presence of a given Rh
gene. Initially, the Rh phenotype was thought to be determined by a single pair of
alleles. However, there are at least eight alleles for the Rh factor. For the purpose
of simplicity, consider here one allele: Rh+ s dominant over Rh-. Thus a person
with Rh+/Rh– genotypes or Rh+/Rh+ has Rh+ blood.
The Genetics of Blood Types
The human blood types A, B, AB and O are inherited by multiple alleles. Multiple
alleles refer to three or more genes that occupy a single locus on a chromosome.
Gene A codes for the synthesis of antigen A on red blood cells, gene B codes for
the production of antigen B on the red blood cells and gene O does not produce
any antigens. The phenotypes listed in the table below are produced by the
combinations of the three different alleles, A, B, O. When genes B and A are
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present in an individual, both are fully expressed. Both A and B are dominant over
O; the genotype of an individual with blood type O must be OO.
TABLE 2
CLUMPING REACTIONS OF ABO BLOOD-TYPING SERA

Phenotypes Possible Genotypes

A AA, AO
B BB, BO
AB AB
O OO

Use A for antigen A; B for antigen B;

O for no antigens present.
Gene A is dominant over O.
Gene B is dominant over O.
AB blood type results when both genes A and B are present.

If we were in lab, you would be doing a synthetic blood typing experiment. But
unfortunately, we are not. So I am giving you data that you can use to answer the
questions below.

BLOOD TYPING DATA SHEET

Patient Name Anti-A Anti-B Anti-Rh Blood Type

Serum Serum Serum
Mr. Smith + + + AB+
Mr. Jones - + + B+
Mr. Green + - - A-
Ms. Brown - - - O-

1. What ABO antigens are present on the red blood cells of Mr. Green’s blood?
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2. What ABO antibodies are present in the plasma of Mr. Green’s blood?

3. If Mr. Jones needed a transfusion what ABO type(s) blood could he safely
receive?

4. If Ms. Brown were serving as a donor, what ABO blood type(s) could receive
her blood safely?

5. Why is it necessary to match the donor’s and the recipient’s blood before a
transfusion is given?

6. What happens to red blood cells that are agglutinated?

7. What is the difference between blood group antigens and blood group
antibodies? (Where is each found?)

8. Explain the basis of ABO blood types. (What is found on the cells of each type
and what is found in the plasma?)
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9. Could a man with AB blood be the father of an O child? Explain.

10 Could a man with an O blood type be the father of an AB child? Explain.

11. Could a Type B child with a type A mother have a type A father? Explain.

12. What are the possible genetic combinations of an offspring when the blood
types of the parents are A and B? Show 4 different Punnett squares to substantiate
your answer.
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13. Suppose Mr. Smith marries Ms. Brown. (Rh+ is dominant.) If Mr. Smith is
homozygous, what are the chances of having a RH+ child? a RH- child?
(Show the Punnett Square.)

If Mr. Smith is heterozygous, what are the chances of having a Rh+ child?
a Rh- Child? (Show the Punnett Square.)

14. Explain how erythroblastosis fetalis may develop.

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15. Under what conditions might a person with Rh- blood develop Rh antibodies?

16. Why can Rh+ blood be given only once to a non-sensitized person who is
Rh-?

17. What is likely to happen to a donor’s cells if a Rh- person who is sensitive to
Rh+ blood receives a transfusion of Rh+ blood?

Part 2: Blood Group Genetics Problems

1. Give the possible genotype(s) for each of the phenotypes below.

PHENOTYPES GENOTYPES(S)
O blood
AB blood
A blood
B blood

2. Show the Punnett Square of a cross between a man who is homozygous for A
blood with a woman who is homozygous for B blood. Give the phenotype(s) and
ratio of the possible children.
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3. Show the Punnett Square of a cross between a woman with AB blood and a man
heterozygous for B blood. Give the phenotype(s) and ratio of the possible children.

4. A man who has type O blood has a child with a woman who has type A blood.
The woman’s mother has type AB blood and her father was type O. What is the
probability that the child has each of the following blood types? Show the Punnett
square.
Type O

Type A

Type B

Type AB

5. A man with A blood marries a woman with B blood. Their child has O blood.
What are the genotypes of these individuals? What other genotypes and in what
frequencies would you expect in offspring from this marriage. Show the Punnett
Square.