CLINICAL APPLICATIONS OF RED CELL IMMUNOLOGY

BLOOD GROUPS
The blood group of a particular individual is the description of the antigenic phenotype of the red
blood cells with respect to a particular system.
Twenty-three blood group systems are recognized
Each system consists of a series of red cell antigens
These antigens are genetically determined by a single locus or very closely linked loci.
Antigens are therefore autosomally inherited, and show marked racial differences in antigenic
frequencies
resence or absence of these antigens may help to determine identity or e!clude paternity
Clinical importance of loo! "ro#p $%$tem$&
"erives from the development of antibodies against these antigens
a. Autoantibodies #against self antigens$
b. Alloantibodies #against foreign antigens$
These antibodies may cause several pathological processes%
a. "estruction of native red cells #Autoimmune haemolytic anemia$
b. "estruction of transfused red cells #&aemolytic transfusion reactions$
c. "estruction of foetal red cells by maternal antibodies which traverse the placental
barrier #&aemolytic disease of the 'ewborn$
Cla$$ification of re! cell antio!ie$
a. I"M % these are capable of complement activation thus causing immediate intravascular
haemolysis
b. I"G& some types can activate complement. All are capable of crossing the placenta
Cla$$ification of re! cell antio!ie$
(. 'aturally occurring %
i. )sually *g+ , but may be *g,
ii. 'ot formed in response to pregnancy or transfusion
iii. -nly clinically important if they are reactive at ./
o
0
1. *mmune
i. redominantly *g,
ii. 2ormed as a result of e!posure to foreign antigen
Ma'or loo! "ro#p $%$tem$&
() ABO
3ithin the A4- system four ma5or blood groups can be recognized%
- 4lood group A #A antigen on 640 7 4 antibody in plasma$
4lood group 4#4 antigen on 640 7 A antibody in plasma$
- 4lood group A4 #A and 4 antigens on rbc 7 No anti A or anti 4
antibodies in plasma$
- 4lood group - #No A nor 4 antigens on rbc 7 anti- A and 4 antibodies in
plasma$
A4- antibodies to antigens that are absent appear in first . months of life.
*)R+,D-
-0omple! system comprising 89 different antigens
- 2ive important antigens% ": 0: E: e: c. 6h #"$ 7ve ; 6h 7ve
6h antibodies are *g,: most cannot fi! complement
.) Ot+er$ ,Minor anti"en$-
'umerous other antigens e.g <ell, "uffy, <idd # not usually tested routinely on donated
blood$
APPLICATIONS OF RBC IMMUNOLOGY
()Bloo! Gro#p Te$tin"
)sually restricted to the A4- and the 6hesus #"$
A$ ABO Bloo! Gro#p
To determine the A4- type, red cells must be tested with anti-A and Anti-4 and the
serum=plasma tested with A and 4 red cells
For/ar! "ro#pin" - identifies the anti"en$ on the red cells
- tests the recipient or donor red cells with anti-A and anti-4 sera
eg 0ells agglutinated only with anti-A serum are group A
0ells that do not agglutinate with anti-A or anti-4 are group -
Re0er$e "ro#pin" > identifies the presence of antio!ie$ in the serum=plasma
- confirms the reaction obtained by the forward grouping test
- tests the serum=plasma from the recipient or donor
with group A red cells and group 4 red cells
eg Agglutination with group 4 cells indicates the presence of
anti-4 in the plasma > ,roup A individual
B$ R+e$#$ Bloo! Gro#p ,R+-
6hesus typing of red cells is determined by e!amining their reaction with anti-" serum.
There are no ?naturally- occurring@ 6hesus antibodies, therefore reverse grouping is not
performed.
6outine testing for other 6h antigens is not required.
*)ANTIBODY SCREENING
Nat#rall% occ#rrin" antio!ie$ occur in the plasma of individuals who lack the corresponding
antigen and who have not been pregnant or transfused. The most important are anti-A and anti-4.
These are usually *g+ antibodies.
Imm#ne antio!ie$ develop in response to the introduction of red cells possessing antigens the
individual lacks. This may occur after pregnancy or previous blood transfusions. These
antibodies are commonly *g, optimally react at ./ 0. -ne e!ample is anti-".
Detection of Imm#ne Antio!ie$
A$ Direct Anti1"lo#lin2Coom$ te$t
Aim% "etects antibody or complement on the re! cell memrane
+ethod%
"ispense (drop of red cell into a test tube
3ash red cells .-8 times with saline #To remove unattached antibodies$
"ecant final wash
Add 0oombs #antihuman globulin$ reagent
0entrifuge
E!amine for agglutination
*nterpretation%
Agglutination indicates a positive test
)ses%
&emolytic "isease of the 'ewborn #&"'$
Autoimmune &emolytic Anemia #A*&A$
"rug-induced *mmune &emolytic Anemia
&emolytic transfusion reactions #&T6$
Aimitation%
A negative test does not necessarily rule out the presence of immune antibodies
B$ In!irect anti1"lo#lin2Coom$ te$t
Aim% "etects antibodies present in the pla$ma
+ethod #1 stage procedure$%
1 drops serum=plasma into test tube
Add, ( drop ,roup - red cells
*ncubate at ./ 0
3ash red cells with saline .-8 times
"ecant final wash
+ethod contBd%
Add 0oombs #antihuman globulin$ reagent
0entrifuge
E!amine for agglutination
*nterpretation%
Agglutination implies that the original serum contained an antibody
)ses% A*&A
&T6
6outine antibody screening of recipientBs serum prior to transfusion
.) Pre tran$f#$ion te$tin" ,T3E CROSS1MATC3-
($ Celect a unit of donor blood that has the same A4- and 6hesus blood group as the recipient
1$ A cross-match must be performed before transfusion of red cells, unless there is an urgent
need for blood.
Aims% "emonstrate A4- incompatibility between donor and recipient
"etect and identify clinically significant immune antibodies in the plasma of the recipient
that will react with donor red cell antigens
4) In0e$ti"ation of +aemol%tic tran$f#$ion reaction$
0ase Ctudy
11 yr -old man with Aplastic anaemia , is on regular transfusions for the past (D months.
Transfused for severe anaemia # &b 8.9 g=dl $and is discharged from hospital
6eturns in 1 weeks with weakness and 5aundice &b is 9.1g=dl
0linical "iagnosis% "elayed &aemolytic Transfusion 6eaction
(. E!posure to minor foreign antigen with previous transfusion
1. 2ormed *g, antibodies #Censitzation$
.. 6e-e!posed to same foreign antigen with the most recent transfusion
8. Antibodies bind to transfused cells causing their destruction in the spleen
Tests for confirming diagnosis
(. *ndirect 0oombs Test
1. "irect 0oombs test
In f#t#re5 all loo! for t+i$ patient m#$t e te$te! for t+i$ minor anti"en an! onl% anti"en
ne"ati0e loo! ma% e #$e!
6) In0e$ti"ation of Neonatal 'a#n!ice
+rs. A is group - 6&#"$>ve . Che is in her .
rd
pregnancy. &er husband is group - 6h#"$ 7ve .
Che delivers a 1.9 kg baby after .D weeks of pregnancy
The baby is severely 5aundiced and has a &b of ((.E g=dl
Clinical Dia"no$i$% &"' # 6h *ncompatibility$
(. +rs A was e!posed to 6h#"$ antigen of her first or second child during pregnancy or
delivery, and formed 6h#"$ antibodies
1. These antibodies crossed the placenta during the .
rd
pregnancy and were bound to
6h#"$ antigens of the foetus
.. 2oetal red cells with antibody were destroyed in the spleen
Tests for confirming diagnosis
(. *ndirect 0oombs Test on maternal serum
1. "irect 0oombs test on infants blood