Basic Immunohaematology

Dr. Bahy Ayoub

MBBCh, MSc, MD, MBA
Red cell Antigens and
Blood Grouping (BG)

First Things First..

Dr. Bahy Ayoub

MBBCh,MSc,MD, MBA
Switzerland
Components of blood
 Q. What determines a blood group?
A. The antigens on the red cell surface.

intro ABO Rh other groups products indications testing dangers

Antigen / Antibody
 ISBT Nomenclature

043
 What are the antigens?

• A, A1, B and AB
• Some people have A antigen (“type A”)
• Some people have B antigen (“type B”)
• Some people have both A and B (“type AB”)
• Some people have neither A nor B (“type O”)

intro ABO Rh other groups products indications testing dangers

Biochemical structure of ABO system
Structure of RBC membrane
 Genotype Antigens Blood type Antibodies

AA
A A anti-B
AO

BB
B B anti-A
BO

AB A and B AB none
anti-A
OO None O
anti-B

intro ABO Rh other groups products indications testing dangers

Isoagglutinins
A sub-groups
 Rh Blood Group System
• The most complex among all blood group
systems.
• Involve in
– Transfusion Reactions
– Haemolytic Disease of Newborn (HDN)
– Autoimmune Haemolytic Anaemia (AIHA)
The Rhesus
System
 The Rh or Rhesus blood system is the second most
important system.
 The major antigen in this system is D

Rh positive ( D+) 85%

Rh negative ( D-) 15%

D Ag is highly immunogenic

Antibodies are "unexpected" and are immune - they

result from previous transfusion or pregnancy
Epitopes Structure
Weak D and Variant D
 Other
Blood groups on the RBC
Luthera ABO Gerbich
n Ii
Knop Dieg
s o
Y
t
Duff
y

MN
S

Crome
r
India LW
n Kel R
l h
Slide courtesy of E. Sjoberg-
Wester
 Blood group antigens as
autoantigens/assciations with diseases
• I/i • Cold agglutinins

•P • Cold agglutinins
• Donath-Landsteiner
• Kell antibodies
• Warm-reactive
autoantibodies
• Lewis
• Helicobacter pylori
– molecular mimicry
• MN • Treponema pallidum
– molecular mimicry

Nydegger UE, Flegel WA 2004

Blood Typing
There are 2 components to blood typing:
• Test unknown cells with known antibodies
• Test unknown serum/plasma with known rbc’s
The patterns are compared and the blood
group is determined.
 FORWARD TYPE

anti-A antibodies AHG

patient
red cells
(type A)

Forward typing is done using both anti-A and anti-B antibodies!

 REVERSE TYPE

reagent AHG
red cells (type B)
patient
serum
(with anti-B Ab)

Reverse typing is done using both type A and type B reagent cells!
Red Blood Cell Antibodies
IgG structure
IgM structure
Reaction of antibodies
Isoagglutinins
Immune Anti-A, Anti-B
Rh HDN
 Antibody Screening

Screening
cells
Antibody Screening, WHY?
Indications: For detection of
irregular antibodies (non-
ABO)
• Pre-transfusion tests
• Antenatal screening
• Donor units
• HDN
• On request,e.g. ICT for
AIHA
Antibody screening procedure
• AHG
• Enzyme
---------------------
• One-cell pool (donors)
• Two cells
• Three cells (recommended)
• Six cells
 Prenatal /Antenatal
Antibody Screening

• Detects all clinically significant IgG alloantibodies that

are reactive at 37ºC.
• If Antibody Screening is Nonreactive, Repeat testing is
recommended at 20 to 24 week’s gestation and again at
delivery.
• At least three separate reagent screening cells, covering
all common blood group antigens, should be used.
• An antibody enhancing medium such as LISS and
Enzyme can increase sensitivity of the assay.
 Antibody Screening

Importance in transfusion
• This test detects if a recipient has a
clinically significant antibody, so a donor
unit that lacks the corresponding antigen
would be selected for XM and transfusion.
Clinical Significance of a Positive
DAT

.
Dr. Bahy Ayoub
MBBCh,MSc,MD. MBA
Switzerland
DAT
 DAT
• The direct antihuman globulin test
(DAT) is needed to demonstrate
antibodies in the event of in vivo
erythrocyte sensitisation.
• Thus, antibodies or complement
components already fixed to the
patient's erythrocytes are detected.
• Following a triple washing
process with the sensitised cells,
the AHG serum is added.
 Causes of Positive DAT
• Incompatible blood • Mixed-Type AIHA
transfusion • PCH
• HDN (due to ABO or • Drug-Induced
other antibodies) • AIHA
• WAIHA • Disease-associated
• CAS • No cause
Clinical Significance of DAT, HDN
 HDN (1)
Risk of Rh D sensitization
 HDN (2)
immune system is alerted by cells transfe
 HDN (3),
antibody production, 1st baby safe
 HDN (4),
immune system antigen recognition
 HDN (5),
Abs. React with following pregnancy
 Rh prophylaxis (1)
Risk of Rh D sensitization
 Rh prophylaxis (2)
anti-D immunization at 28 weeks
 Rh prophylaxis (3),
cells are coated with Rhogam
Rh prophylaxis (4),
the mother is safe..
THANK YOU