IMMUNOHEMATOLOGY

JQGACASAN, MD
August, 2016

www.slideshare.net
Immunohematology
• Study of immunologic reactions of all
components of blood

• Antigens, antibodies, antigen-antibody

reactions
Applications
• transfusion of blood and its components
• diagnosis, prevention, & management
of immunization associated with
pregnancy
• leucocyte testing for organ
transplantation
• resolution of parentage problem
ABO Blood Group
ABO Blood Group System
• Single most important blood group for
selection and transfusion of blood

• Major consideration in solid organ and

bone marrow transplantation

• Widely expressed (blood, tissues, fluids)

ABO GENETICS
PHENOTYPE GENOTYPE Ag Ab

A AA, AO H, A Anti B

B BB, BO H, B Anti A

AB AB H, A, B None

O OO H Anti A and
Anti B
ABO Blood Group Antibodies
• Most important Abs in transfusion medicine
• Generally, naturally occurring
• Weak or absent in:
• Sera of newborns ‘til 3-6 months
• Adult levels at 5-10 years of age
ABO Blood Group Antibodies
• naturally-occurring
• mostly IgM
• absence of stimulation by either
transfusion or pregnancy
• detected at room temp using saline
agglutinins w/ optimal reactivity at
4˚C
Most anti A and anti B are IgM,
type “O” individuals in addition have IgG anti A and B
ABO Antibodies
IgG antibodies are reactive at 37C
• Follows immune stimulation by
transfusion or pregnancy

• Can cause hemolytic disease of the

newborn and acute rejection of
transplanted organ
ABO Blood Group NULL
Phenotypes
Classic Bombay (Oh)
• neither AB nor H antigens present on red cells or
in secretions
• lack Se gene (se/se)
• anti A, anti B and anti H are present in sera
Para-Bombay
• little or no ABH expression on red cells
• some with normal expression of ABH Ags in
secretions & body fluids
Frequencies (%)

Blood White Black Native Asian

type America
n
O 45 49 79 40
A 40 27 16 28
B 11 20 4 27
AB 4 4 <1 5
ABO Blood Group Detection
1. Forward typing –
patient red cells with reagent anti A and
anti B

2. Reverse typing -
patient serum or plasma with A and B
red cells
ABO Blood Group Detection
Anti A Anti B A cells B cells
“A” (+) (-) (-) (+)
“B” (-) (+) (+) (-)
“AB” (+) (+) (-) (-)
“O” (-) (-) (+) (+)

+ = Agglutination
Anti-sera for forward typing
Red cells for reverse typing

www.bio-rad.com
Forward Typing
Hemolytic Disease of the Newborn
(ABO)
• mother - “O”
• fetus not “O”
• first baby
• very common
• very mild
On the lighter side…
en.wikipedia.org
randomcoolstuff101.blogspot.com
Rh System
• Transmission follows autosomal dominant
pattern

• D antigen – most immunogenic

• Followed by c, E, C, and e
medcaresupplies.ph

• gene RhD & RhCE determines the D, C, c, E, e

Ag
• “Rh positive” with D antigen
• “Rh negative” no D antigen
HEMOLYTIC DISEASE OF THE NEWBORN (Rh)

• Rh negative (-) mother develops

anti D following exposure to the
Rh-positive (D cells) of the fetus

• Pregnancy and childbirth are the

immunizing events
Rh antibody stimulation
Hemolytic Disease of the Newborn
(Rh)
• mother Rh negative
• fetus Rh positive
• usually second baby
• uncommon
• severe
– Hydrops fetalis
Immunohematology
Tests and Procedures
• Hemaglutination (basic principle) 2 stages:

1. RBC sensitization – combination of paratope

& epitope

2. Multiple RBCs w/ bound Abs form a stable

latticework thru Ag-Ab bridges formed bet
cells
Normal (+) agglutination
Grading of hemagglutination
• Hemolysis – strongest positive reaction
• One solid aggregate (4+)
• Several medium to large aggregates
(3+)
• Many small to medium aggregates w/ a
clear background (2+)
• Many small aggregates w/ a turbid
background (1+)
AntiGlobulin / Coombs Tests
• Antibody directed against human IgG (AHG)

• Direct Antiglobulin Test (DAT)

• AHG detects the presence of antibody or
complement molecules on RBC surface

• Indirect Antiglobulin Test (IDAT)

• AHG detects the presence of antibodies in serum
Other blood group systems
• Lewis
•P
•I
• MNS
• Lutheran
• Kell
• Duffy
• Kidd
Thank you!