BACHELOR OF BIOMEDICAL SCIENCES

BASIC TRANSFUSION SCIENCE AND BLOOD BANKING

BMC 125

CHAPTER 4: RH TYPING SYSTEM

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LEARNING OUTCOMES

At the end of the lecture, students should be

able to:
• Discuss the Rhesus system
• Discuss nomenclature of Rh system
• Describe Rh antigen, Rh antibody, Rh null,
weak ‘D’ antigen.

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 The Rhesus System
• The Rhesus system (Rh) was originally described in
1939 by Landsteiner & Wiener.
• Antibodies produced in animals injected with blood
from Rhesus monkeys were found to agglutinate red
cells from ~ 85% of humans.
• The Rhesus monkeys and humans shared a red cell
antigen system – the Rhesus system.
• Approximately 85% of the UK population are Rh (D)
positive.

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 The Rhesus System
• The study of subsequent Rh antibodies produced in
humans showed that many contained more than one
antibody specificty.
• This led to the discovery that other Rh antigens
existed, in addition to D.
• Other antigens included C,c, E and e.
• The Rh system is one of the most complex multi-allelic
blood group antigen systems known.
• The genes encoding the Rh system are located on
chromosome # 1 (the ABO genes are on #9)

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 Rh Antigen Frequency
• D antigen – 85%
Rh Positive
• d antigen – 15%
• C antigen – 70% Rh Negative
• c antigen – 80%
• E antigen – 30%
• e antigen – 98%

• The presence or absence of D Ag determines if the person is

Rh+ or Rh-

Mohammed Laqqan
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Nomenclature of the RH system

Different nomenclatures:
1- Fisher-Race
2- Weiner

Mohammed Laqqan
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 Fisher-Race Theory
• Rh inheritance is controlled by 3 closely linked loci on each
chromosome of a homologous pair
• Each locus has its own set of alleles which are: Dd , Cc , and
Ee .
• The D gene is dominant to the d gene, but Cc and Ee are co-
dominant.
• The 3 loci are so closely linked, and the 3 genes on one
chromosome are always inherited together.

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• In this theory either of the alternative alleles
at each of the three Rh gene loci are inherited
from each parent i.e as a ‘triplet’

Maternal chromosome D C e

Paternal chromosome
d cc ee

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• In this genetic model the ‘d’ gene is considered as an
amorphic or silent gene, as no antigenic product has
been described.
• The d gene is assumed to exist as an allelic partner to
the D gene.
• The other five genes are co-dominant, with each
producing a demonstrable product, which will initiate
antibody formation against it.

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• A person may produce various combinations
of these antigens, dependent on which
genes they inherit from each parent.
• In the example given the offspring will be
CDe/cde – the possession of the D antigen
means that the offspring will be classed as
Rhesus +ve

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 Fisher-Race

• There are 8 haplotype gene

complexes at the Rh locus DCe dCe
• Fisher-Race uses DCE as DcE dCE
the order
• Others alphabetize the Dce dcE
genes as CDE DCE dce
• 8 haplotype gene
complexes are possible
from the three loci.

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 Wiener Theory
• Good for describing phenotype
• There is one Rh locus at which occurs one Rh gene,
but this gene has multiple alleles.
• For example, one gene R1 produces one agglutinogen
(antigen) Rh1 which is composed of three "factors"
• The three factors are analogous to C, D, and e
respectively
• The main difference between the Fisher-Race and
Wiener theories is that the:
– Fisher-Race theory has three closely linked loci,
– the Wiener theory has only one gene locus at which
multiple alleles occur.
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 Conversion of Wiener to Fisher-Race

• R in Wiener = D in Fisher-Race
• r is absence of D (d)
• 0 or no symbol implies c and e
• 1 or ′ implies C and e
• 2 or ″ implies c and E
• z or y implies C and E

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Fisher-Race and Wiener Nomenclature

)Weiner Gene( Antigens Fisher-Race

R0 D, c, e Dce
R1 D, C, e DCe
R
2
D, c, E DcE
R
z
D, C, E DCE
r c,e dce
′r C,e dCe
″r c,E dcE
ry C,E dCE

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 Rh Nomenclature
• Each of these haplotypes has been given a shorthand
notation.
• The use of ‘R’ indicates the presence of the D gene
whereas the use of ‘r’ indicates D absence (i.e d/d)
• All combinations possessing at least one D gene are
Rhesus positive.
• All combinations lacking the D gene are Rhesus
negative.

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 Rh Nomenclature

• To illustrate:-
– cde/cde combination is also rr
– CDe/cde combination is also R1r
– cDE/cdE combination is also R2r’’
• Certain combinations of genes are more
common than others, and hence some of the
genotypes are relatively common whereas
others are very rare.

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 Rh haplotype frequencies

• The frequency of a particular haplotype also

varies between different populations/races,
e.g. Rh negative (rr) is extremely rare in
Chinese populations.
• In negroid populations the haplotype cDe (R0)
is more common than in Caucasian
populations.

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 Rh Grouping

• When red cells are grouped using specific

antisera against Rh antigens the combination
of antigens identified is known as the
phenotype. E.g:-
Reactions with antisera Phenotype
Anti-C Anti-D Anti-E Anti-c Anti-e

+ + - + + CcDe
- + + + + cDEe
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 Rh Grouping
• To illustrate with our examples:-

Phenotype Possible Genotypes % Frequency of

genotype
CcDe CDe/cde R1r 35
CDe/cDe R1R0 2
cDe/Cde R0r’ <0.1

cDEe cDE/cde R2r 12

cDE/cDe R2R0 0.7
cDe/cdE R0r’’ <0.1

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 Rh Antigens

• These antigens, coded for by the Rh genes are non-

glycosylated, non-phosphorylated transmembrane
proteins.
• The Rh antigens are made up of 417 amino acids.
• The C and c antigens differ by only 4 amino acids.
• The E and e only differ by a single amino acid
substitution

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 Rh Antibodies
• The vast majority of Rh antibodies are of the IgG
class.
• Rh antibodies are immune and react optimally at
37ºC.
• Rh antibodies do not activate complement due to
relatively sparse distribution of antigen molecules
per cell.
• They are still capable of causing the destruction of
incompatible red cells in vivo, by removal of coated
red cells by the spleen macrophages.

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 Rh null
• RH null: individual that appears to have no Rh
antigens
• RBC has fragile membrane- short lived
• Must use autologous blood products
– No D, C, c, E, e antigens present on the RBC
membrane
• Demonstrate mild hemolytic anemia (Rh antigens are
integral part of RBC membrane and absence results
in loss of membrane integrity)
– Stomatocytosis.
– When transfusion is necessary ONLY Rh Null
blood can be used to transfuse.
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 Rh antibodies

• Result from the exposure Rh Abs

to Rh antigens Clinically Abs class
• IgG form Significant IgG
• Bind at 37°C Yes

• Form agglutination in IAT Thermal range HDNB

37 - 4 Yes
phase
Transfusion Reactions
Extravascular Intravascular

Yes No
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 Weak D Phenotype

• Most D positive rbc’s react macroscopically with Reagent

anti-D at immediate spin
– These patients are referred to as Rh positive
– Reacting from 1+ to 3+ or greater

• HOWEVER, some D-positive rbc’s DO NOT react (do

NOT agglutinate) at Immediate Spin using Reagent Anti-D.

These require further testing (37oC and/or AHG) to

determine the D status of the patient.
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Variants of D

• Weak expression of the Rh system on the

RBC, (Du)
• Du red cells can be classified into different
categories according to the mechanism that
account for the Weak D antigen

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 Categories of D red cells
u

1- Du Variant (Partial D)
2- Hereditary Du (Genetically Transmissible)

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 1- D Variant (Partial D)
u

• The D- Ag consists of at least 4 parts

• Missing one or more PARTS (epitopes) of the D
antigen remaining Ag is weakly expressed
• Alloantibodies are produced to the missing parts
• Du variants should receive Rh –ve blood when
transfused

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 Partial D: Multiple epitopes make up D antigen. Each color
represents a different epitope of the D antigen.

A.

Patient B lacks
B. one D epitope.
The difference between Patient A and Patient B is a single epitope of the D
antigen. The problem is that Patient B can make an antibody to Patient A even
though both appear to have the entire D antigen present on their red blood cell’s
using routine anti-D typing reagents.. BIOMEDICAL SCIENCE, FACULTY OF MEDICINE
 2- Hereditary Du (Genetically Transmissible)

• The RHD gene codes for weakened expression of D antigen

in this mechanism.
– D antigen is complete, there are just fewer D Ag sites on the rbc.

– Common in Black population (usually Dce haplotype). Very rare in

White population.
• Agglutinate weakly or not at all at immediate spin phase.
• Agglutinate strongly at AHG phase.
• Can safely transfuse D positive blood components.

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 REFERENCES
• Denise M. Harmening PhD MLS(ASCP)
CLS(NCA),2012.Modern Blood Banking &
Transfusion Practices 6th Edition.by F.A.Davis
company united sate of America.
• Eva D.Quinley Ms MT
(ASCP)SBB.2010.immunohaematology Principles and
practice,3rd ed( Lippincott Williams and Wikins)
• Jeffrey Mc Cullough.2011.Trnasfusion Medicine 3 rd
ed,wiley Blackwell publication ,United states.
• Harvey G.Klein,David J Anstee 2013,Mollisons blood
transfusion in clinical medicine,12 th ed,Wiley-Blackwell.
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Thank you

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