Other Blood

Group Systems
Part 4

MLS 404
ISBT TERMINOLOGY
001 - 036 (<100) series for BGS
200 series for BGC
700 series for low prevalence
antigens
901 series for high prevalence
antigens
Terminologies
 Blood Group Systems - group of antigens
produced by alleles at a single gene locus
or loci so closely linked that crossing over
does not occur or is very rare
 Most blood group genes are located on the
autosomal chromosomes, EXCEPT
 Xg Blood group system (ISBT 012) Xp22.33

 Kx Blood group system (ISBT 019) Xp21.1

 Found in X chromosome (more

common in females
Terminologies

 Blood Group Collection - serologically,

biochemically, or genetically related
antigens; not necessarily produced by
alleles at a single gene locus or loci.
Terminologies
 Codominant - a pair of genes in
which neither is dominant over the
other; both expressed
 Antithetical - antigens that are
product of allelic genes; opposite
form of a gene
 Amorph - gene that does not
appear to produce a detectable
antigen; a silent gene (e.g. O
gene)
Terminologies
 Clinically Significant Antibodies
- antibodies reactive at 37 degree Celsius
(warm reactive)
- usually IgG
- causes transfusion reactions

 Clinically Insignificant Antibodies

-cold reacting (RT) antibodies
- usually IgM
- rarely causes complications in blood
transfusion
- "NUISANCE ANTIBODIES"
ALWAYS Clinically Significant Blood
Group Antigens
A and B
 H in Oh

 Rh

 Kell

 Duffy

 Kidd

 Diego

 S, s, U (MNS)

 P, PP1 Pk (P1Pk)

 Vel (BGC)
SOMETIMES Clinically Significant Blood
Group Antigens
 Ata (high incidence Ag)
 Colton
 Cromer
 Dombrock
 Gerbich
 Indian
 Jra(high incidence Ag)
 JMH
 Lan (high incidence Ag)
 Landsteiner - Wiener
 Scianna
 Yt
Not Clinically Significant Blood Group
Antigens UNLESS REACTIVE AT 37°C

 A1 (ABO)
H (ISBT 018)
 Lea (Lewis)
 Lutheran
 M, N
 P1 (P1Pk)
 Sda (high incidence Ag)
NOT USUALLY Clinically Significant
Blood Group Antigens

 Chido / Rodgers (ISBT 017)

 Cost (BGC, 200 series)
 Knops
 Leb (Lewis)
 Xga (Xg)
Clinically Significant Antibodies
 Anti-Rhesus (D, C, c, E, e)
 Anti-Kell (K)
 Anti-Kidd (Jka, Jkb)
 Anti-Duffy (Fya, Fyb)
 Anti-s
 Anti - A and Anti - B
Clinically Non-Significant Antibodies

 Anti-Lewis (Lea, Leb)

 Anti-M
 Anti-N
 Anti-S
 Anti-P1
 Anti-H in A1 and A1B
Cold Antibodies (IgM) Warm antibodies (IgG)
 Anti-Lea  Rh
 Anti-Leb  Kell
 Anti-I
 Duffy
 Anti-P1
 Kidd
 Anti-M
 Anti-A, -B, -H  Anti-S,-s,-U

 Anti-N  Anti-Lub
 Anti-Lua
Remember enzyme activity:
Enhanced Destroyed
by enzymes by enzymes
Papain
Kidd Fya and Fyb
Bromelin,
Rh M, N
Ficin
Lewis S, s
Trypsin
I
P
Dosage Effect:
 Kidds and Duffy the Monkey (Rh) eat lots of M&Ns

M&Ns
M&Ns

anti-Jka, -Jkb, -Fya, -Fyb, anti-C, -c, -E, -e (no D), -M, -N, -S, -s
MNSs
Kidd Duffy Rh

adapted from Clinical Laboratory Science Review: A Bottom Line Approach (3rd Edition)
Lewis (ISBT 007)
 ISBT007 (Lea, Leb); ISBT 210 (Lec, Led)
 discovered by Mourant (1946)

 produced by tissue cells (intestinal

epithelial cells)& ADSORBED ONLY TO
RBC MEMBRANES
 Le gene (FUT3) codes for the
production of fucosyltransferase
enzyme
Lewis (ISBT 007)
 ISBT007 (Lea, Leb); ISBT 210 (Lec, Led)
 discovered by Mourant (1946)

 produced by tissue cells (intestinal

epithelial cells)& ADSORBED ONLY TO
RBC MEMBRANES
 Le gene (FUT3) codes for the
production of fucosyltransferase
enzyme
 SUBSTANCE RED CELL
GENOTYPE Le Abs
(SECRETION) PHENOTYPE

ABH
ABH, lele, sese None Anti-Lea, Anti-Leb
le (a-b-)

ABH, lele, SeSe ABH

ABH Anti-Lea, Anti-Leb
or Sese Le (a-b-)

ABH, LeLe or ABH

(Lea Anti-Leb
Lele, sese Le (a+b-)

ABH, LeLe or ABH

Lele, SeSe or ABH Lea Leb None
Sese Le (a-b+)
Lewis Phenotype Frequencies

PHENOTYPE WHITES (%) BLACKS (%)

Le (a+ b-) 22 23

Le (a-b+) 72 55

Le (a-b-) 6 22

Le (a+b+) rare rare

Lewis Phenotypes
 Le (a-b-)
 RARE; common in Blacks and pregnant women
 Seen also in newborns <10days old (cord blood)
 Le(a+b-)
 Develop after about10 days in newborns
 Seen in Polynesians, Japanese or Taiwanses

 Le(a+b+)
 Rare; Not a true phenotype; seen only in newborns
during transition (development) of antigens
 Le(a-b+)
 MOST COMMONLY ENCOUNTERED
 72% in Whites & 55% in Blacks
Lewis Antigens
- Cord blood and red cells from newborn
infants phenotype as Le (a-b-)
-May take 6 years to develop
-Decrease in expression of Lewis antigens
has been demonstrated on red cells from
many pregnant women
- not intrinsic to RBCs but passively
adsorbed onto the RBC membrane
from the plasma
Lewis Antibodies
(Anti-Lea and Anti-Leb)
Antibody Class: IgM
Optimal reaction temperature:
37ºC and 4ºC
Reactive phase: 37ºC and AHG
Enzyme treatment: enhanced
(increased agglutination
Alleles: Le and le (amorph)
 Lewis Antibodies

- Considered naturally occurring

Activate the complement and can
cause in vivo and in vitro hemolysis
Agglutination is WEAK, DIRTY LOOKING

Anti-Lea

 most commonly encountered

Disease Association
 H,Ley and Leb antigens are receptors for H.
pylori
 Increased incidence of ulcers and stomach
cancer among blood group O secretors

 Sialyl-Leaand Sialyl-Lex are ligands for the

endothelial adhesion molecule E-selectin
and may mediate tumor cell–endothelium
interactions.

 Sialyl-Lea/Sialyl-Lex
is also the epitope for the
t u m o r m a r k e r C A 1 9 - 9 found in
gastrointestinal and other malignancies
MNS (ISBT 002)
 discovered in 1927 by Landsteiner and Levine
 named from second and fifth letter of the word
"immune"
 S discovered in 1947 by Walsh and
Montgomery, named after Sydney city where
first anti-S was discovered
 s discovered in 1951

 U discovered in 1953 by Weiner (stands for

almost "universal" distribution)
 En in Ena stands for "envelop“
 M-N- cells are also Ena- →has anti-Ena
MN Antigens
 Well developed at birth
 Found on Glycophorin A (MN-
sialoglycoprotein)
 the major RBC sialic acid–rich glycoprotein
 MN Antigens differ in their amino acid
residue at positions 1 and 5:
-M has serine and glycine at these
positions
-N has leucine and glutamic acid
MN Antigens
 Important
for paternity testing of infants
and young children

 Easily
destroyed or removed by
enzymes

 P.falcifarum uses GPA and GPB as

alternative receptors for cell invasion
Ss Antigens
 Located on Glycophorin B (Ss-
sialoglycoprotein)
 Amino acid at position 29 on GPB is
critical to antigen expression
 S has methionine, whereas s has
threonine
 Well developed at birth

 Less easily degraded by enzymes

Anti-M
 Most examples are naturally occurring, cold
reactive saline agglutinins
 IgM or IgG antibody
 Usually do not bind complement
 Do not react with enzyme-treated cells
 Some examples are pH-dependent at pH
6.5
 Other examples react only with red cells
exposed to glucose solutions
 Rarely causes hemolytic transfusion
reactions or HDN
Anti-N
 Cold reactive IgM or IgG saline
agglutinin
 Does not bind the complement

 Implicated only with rare cases of

HDFN
 Seen in renal patients who are
dialyzed on equipment sterilized with
formaldehyde (Anti-N like Ab or Anti-
Nf)
 Anti-S, Anti-s, and Anti-U
- Clinically significant; causes HTR and HDFN
- Anti-U is rare and occurs in S-s- people

NOTE:
M-N-Ena (-) Enveloped Ag
S-s-U (-) Universal
-in BLACKS producing anti-U

Anti-U discovered in 1953 by Weiner (stands

for the almost “universal” distribution of
the new blood factor)
 Miltenberger Series
- low frequency antigens of the MNS
blood group system

PHENOTYPE WHITES (%) BLACKS (%)

M+N- 28 26
M+N+ 50 44
M-N+ 22 30
S+s- 11 3
S+s+ 44 28
S-s+ 45 69
S-s-U- 0 <1
Mk Phenotype

 The
RBCs of these individuals typed
M–N–S–s–U–En(a–)Wr(a–b–)

 Mkgene represents a single, near-

complete deletion of both GYPA
and GYPB

M k M k represent the genotype of

null phenotype in the MNS system
M and N Lectins
M reactivity
- Iberis amara - Iberis umbellate
- Iberis semperivens - Japanese turnip
 N reactivity
- Bauhinia varriegata - Bauhinia purpura
- Bauhinia candicans - Vicea graminea
- Bauhinia bonatiana

Note: Madura aurantiaca (Osage orange)

has both M and N reactivity
 P (ISBT 003) & Pk (ISBT 209)
P1 antigen
 Found on fetal red cells as early as 12 weeks,
but it weakens with gestational age
 Deteriorates rapidly on storage

 P1-like antigen has been found on plasma,

and droppings of pigeons and turtledoves,
as well as in the egg white of turtledoves
 P1 substance has been identified in hydatid
cyst fluid extracts of Lumbricoides terrestris
(common earthworm) and Ascaris suum
P null
p phenotype
 Very rare, 5.8 in a million
 Slightly more common in Japan,
North Sweden, and in an Amish
group in Ohio
 Produces anti-PP1Pk (formerly anti-
Tja)
 DETECTABLE POSSIBLE
PHENOTYPE
ANTIGENS ANTIBODIES

P1 P, P1 none
P2 P Anti-P1
Anti-PP1Pk or
p none
Anti-Tja
P1k Pk, P1 Anti-P

k Anti-P and
P2 Pk
Anti-P1
Frequency of P Phenotypes

PHENOTYPE WHITES (%) BLACKS (%)

P1 79 94

P2 21 6

p rare rare

P1k very rare very rare

P2k most rare most rare

Anti-P1
 Common, naturally-occurring IgM
antibody in the sera of P2 individuals
 Cold-reactive saline agglutinin

 Stronganti-P1 was observed in individuals

infected with Echinococcus granulosus
(Hydatid disease)

 Associated with parasites

 fascioliasis, Clonorchis senensis & Opistorchis
viverrini infections
Anti-PP1Pk
 Originally called Anti-Tja
 Trivia: Mrs. Jay, T= “tumor”
(adenocarcinoma)

 Predominantly IgM, sometimes IgG

 Reacts over a wide thermal range
 Associated with spontaneous abortions in
early pregnancy
 Anti-P
 Naturally occurring alloantibody in the sera
of all Pk individuals
 Alloanti-P is rarely seen in the blood bank but
very significant in transfusion because it is
hemolytic with a wide thermal range of
reactivity
 Anti-P specificity is found as an IgG
autoantibody in patients with Paroxysmal
Cold Hemoglobinuria
 Antibody activity is biphasic - attaches to red
cells in cold and lyses as they warm
 Demonstrated by the Donath-Lansteiner Test
Donath-Landsteiner Test

Control Test
(px WB) (px WB)

30 mins 37 deg C 4 deg C

30 mins 37 deg C 37 deg C

Normal : NO HEMOLYSIS on BOTH tubes

PCH : CONTROL TUBE has NO HEMOLYSIS
TEST TUBE has HEMOLYSIS
Invalid : HEMOLYSIS on Control tube
Anti-Pk
 Isolated from some examples of anti-
PP1Pk by selective adsorption with P1
cells
 Has been reported in the serum of P1
individuals with biliary cirrhosis and
autoimmune hemolytic anemia
I Blood Group System (ISBT 027) and
blood group collection 207 (i)
 discovered in 1956 by Weiner and
colleagues in a patient with CHAD (Cold
Hemagglutinin Disease)
 used the symbol I to emphasize the
"individuality" of RBCs failing to react with
patient's serum at room temp
 Can be a bothersome (nuisance) antibody.
 May mask the reactions of clinically significant
alloantibody.
 May need prewarm cell suspension and reagent to
find clinically significant alloantibodies.
I and i Antigens
 Both I and i are high-prevalence antigens

 Infant RBCs are rich in i which gradually

converts to I during the first 18 months of
life, thus adult RBCs are rich in I and have
only trace amounts of i antigen.

I and i antigens are precursors for the

synthesis of ABO and Lewis antigens
I- phenotype (Adult i)

 Individuals who do not change their

i status after birth
 Mayproduce alloanti-I if transfused I+
blood

↑ i Ag = HEMPAS (Hereditary
Erythroblastic Multinuclearity with Positive
Acidified Serum)
Alloanti-I
 Exists as an IgM or IgG antibody in the serum of
most individuals with the adult i phenotype.
 NOT associated with HDFN because the
antibody is IgM, and the I antigen is poorly
expressed on infant RBCs
Autoanti-I
 Benign Autoanti-I
-Found in the serum of many normal healthy individuals
-Not associated with in vivo red cell destruction
-Weak, naturally occurring, saline-reactive IgM
agglutinin
-Usually reacts only at 4 degree Celsius
 Pathologic Autoanti-I

-Potent IgM agglutinin with higher titer and broader

thermal range of activity, reacting up to 30 or 32 deg
Celsius
-Attach in vivo and cause autoagglutination and
vascular occlusion (Raynaud's phenomenon) or
intravascular hemolysis
Pathologic Autoanti-I
 Production of autoanti-I may be stimulated
by microorganisms carrying I-like antigen on
their surface
 Patients with Mycoplasma pneumoniae often
develop strong cold agglutinins with I
specificity as a cross-reactive response to
Mycoplasma antigen
 Listeria monocytogenes organism from a
patient with cold autoimmune hemolytic
anemia has been reported to absorb anti-I
and stimulate its production in rabbits
Anti-i
 Most autoanti-i are IgM
 Reacts best with saline-suspended cells at
4 deg Celsius
 potent examples are associated with:
Infectious Mononucleosis
Alcoholic cirrhosis
Myeloid leukemia
Reticuloses
Note: IgG anti-i has also been described
and associated with HDFN
Note:
↑I ↑i
Adult Cells Cord cells
strong
A. Anti-I - / weak
reaction
strong
B. Anti-i - / weak
reaction

✓ Cold Agglutinin Disease &

Primary Atypical Pneumonia → Anti-I (IgM)
✓ Paroxysmal Cold Hemoglobinuria
→ autoanti-P (IgG)
Kell (ISBT 006)
 Antigens
K - Kell
k - Cellano
Kpa - Penney
Kpb - Rautenberg
Jsa - Sutter
Jsb - Matthews
Kell Antigens
 Have disulfide-bonded regions on glycoproteins
 Sensitive to sulfhydryl reagents: 2-mercaptoethanol
(2-ME), Dithiothreitol (DTT), 2-
aminoethylisothiouronioum bromide (AET)
 Glycine-acid EDTA (an IgG-removal agent) also
destroys Kell antigens.
 K antigen is the SECOND MOST IMMUNOGENIC
antigen (after D) ; can cause HTR and HDFN
 Kellnull : RBCs that lack Kell Ags but have the Kx
Antigen (K0)
- have the greatest amount of Kx
Kell Antigens
 Kell
antigens are found on kell
glycoprotein which is covalently linked to
XK protein by a single disulfide bond

 Kell
antigen expression is dependent
upon the presence of the XK protein

 Absence of XK protein =
McLeod phenotype
 Absence of Kell antigens = Ko
McLeod Phenotype
 Individual lacks/depressed expression of Kell
and no Kx
 Discovered in 1961 by Allen and co-workers in
a student (Mr. McLeod)
 Decreased RBC survival & RBC morphologic
and functional abnormalities (acanthocytosis)
as well as muscular and neurologic defects
including muscle wasting, elevated CPK,
psychopathology, seizures, cardiomyopathy
which develops on 4th decade of life (Mcleod
Syndrome)
 McLeod Syndrome is assoc. with Chronic
Granulomatous Disease
Kellnull(KoKo) and Anti-Ku
 Ko RBCs lack expression of all Kell
antigens but have the Kx antigen
(found on XK protein)

 If
immunized, produce an antibody
called anti-Ku which recognizes the
“universal” Kell antigen (Ku) present
on all RBCs except Ko.
Frequencies of Common Kell
Phenotypes
PHENOTYPES WHITES (%) BLACKS (%)
K-k+ 91.0 96.5
K+k+ 8.8 3.5
K+k- 0.2 <0.1
Kp (a+b-) <0.1 0
Kp (a+b+) 2.3 Rare
Kp (a-b+) 97.7 100
Js (a+b-) 0 1
Js (a+b+) Rare 19
Js (a-b+) 100 80
Anti-K
 Outside the ABO and Rh antibodies,
anti-K is the most common antibody
seen in the blood bank.
 Anti-K is usually IgG and reactive in
the antiglobulin phase and can
cause HTR and HDFN
 Since K antigen is well expressed on
erythroid cells, Kell HDFN is
characterized by destruction of both
circulating and immature red cells in
the bone marrow
Kx (ISBT 019)
 Kx antigen is in the Kx system
 Antigen is inherited independently from Kell
antigens but is biochemically similar
 K0 (Kell null) individuals have increased
amounts of Kx
 Kx is present in all RBCs except those with
McLeod Phenotype
 XK gene →located in the short arm of X
chromosome
Duffy (ISBT 008)
 Discovered in 1950 by Cutbush and
associates
 Named after Mr. Duffy, a multiply
transfused hemophiliac with Anti-Fya

 Duffygene : 1st human gene to be

assigned to a specific chromosome
(Chromosome 1)
Fy (a-b-)
 MostAfrican-Americans are Fy(a-b-)
 Fy(a-b-) discovered in 1955 by
Sanger and colleagues from African
blacks
(example of natural selection: this phenotype
makes them resistant to malarial infection
[Plasmodium knowlesii and later, P. vivax] which is
endemic in their region)
 Frequency of Duffy Phenotypes

AMERICAN CHINESE
PHENOTYPE WHITES (%)
BLACKS (%) (%)

Fy (a+b-) 17 9 90.8

Fy (a+b+) 49 1 8.9

Fy (a-b+) 34 22 0.3

Fy (a-b-) Very rare 68 0

Duffy Antigens
 Antigens:
SIX: Fya, Fyb, Fy3, Fy4, Fy5, Fy6
 Fyb discovered by Ikin and coworkers in 1951
from a multiparous female (gravida 3 para 3)
 Fy3 – determinant common to Fya and Fyb
 Fy5 – results from the interaction of Rh and Duffy
genes. Not present on Rhnull RBCs
 Fy6 antigen – receptor for P. vivax

 Identified on fetal red cells as early as 6 weeks

gestational age and are well developed at birth
 Destroyed by common proteolytic enzymes
Anti-Fya and Anti-Fyb
 Usually IgG and react best at the AHG
phase
 Enhanced in LISS

 Do not react with enzyme-treated cells

 Associated with HTR and HDFN

 Weak antibodies react stronger with

homozygous cells (dosage effect)
 Anti-Fy3:
 found in the serum of an Fy(a–b–)
individuals
NOTE!

 Anti-Fya
(high incidence Ab) is
more common than Anti-Fyb

 Therefore,
Fyb is a high incidence
Ag as compared to Fya
Kidd (ISBT 009)
 Discovered in 1951 by Allen and
coworkers

 Named after John Kidd, the sixth child

of Mrs. Kidd, who has HDFN

 Jkb
discovered in 1953 by Plaut and
associates
Kidd null phenotypes
 In(Jk)
– has inhibitor to Jk but has no Ab
against Jk

 Jk
(a-b-) described in 1959 by Pinkerton
and associates
 Null phenotype of Kidd BGS
 Produces anti-Jk3:
 most abundant among Polynesians,
Filipinos, Indonesians, Chinese and
Japanese.
Frequencies of Kidd Phenotypes

PHENOTYPE WHITES(%) BLACKS(%) ASIANS(%)

Jk (a+b-) 28 57 23

Jk (a+b+) 49 34 50

Jk (a-b+) 23 9 27

Exceedingly Exceedingly
Jk (a-b-) 0.9 - <0.1
rare rare
Kidd Antigens
 Jka and Jkb antigens are well developed
on the RBCs of neonates.
 RBCs carry 14,000 antigen sites per cell
 Enhanced by enzymes
 Jk null RBCs are resistant to lysis by 2M
urea, a lytic agent used by some
automated hematology analyzers.
 Found on Kidd glycoprotein which is a
urea transporter (multipass)
Kidd Antibodies
 Demonstrate dosage
 Difficult to detect.
 Weak, titer of anti-Jka or anti-Jkb quickly declines in
vivo.
 Found in combination with other antibodies
 These antibodies bind the complement
 These antibodies are associated with
HTR and HDFN
 The decline in antibody reactivity and
the difficulty in detecting Kidd
antibodies have a notorious reputation
in blood bank as a common cause of
delayed HTR
MUST KNOW!
 Demonstrates dosage effect
 Drug-induced HTR: in patients receiving
Aldomet / Methyldopa (source: Harmening)
 Enterococcus faecium and Micrococcus
can convert Jk(b-) to Jk(b+) (transient)
 Proteus mirabilis is a stimulus for
autoanti-Jkb
LUTHERAN (ISBT 005)
 Discovered in 1945; named after a patient
named Lutteran, who has lupus erythematosus
diffusus, who underwent multiple transfusions

PHENOTYPES WHITES(%) BLACKS(%)

Lu (a+b-) 0.15 0.1

Lu (a+b+) 7.5 5.2

Lu (a-b+) 92.35 94.7

Lu (a-b-) Very rare Very rare

Lua and Lub Antigens

 Poorlydeveloped at birth
 Do not reach adult levels until age 15

 Causes only mild HDFN

Lu NULL PHENOTYPES:
 Lu (a-b-)
 recessive type of Lu null wherein px did not

inherit any of the Lu antigens (produces the

anti-Lu3
 In (Lu)
 Inheritance of Lu but an “inhibitor” is present;

px does not produce anti-Lu

Anti-Lua
 naturally-occurring
 Mostly IgM, cold reacting
Few react at 37 deg by indirect
antiglobulin test
May be IgA, IgM, or IgG
Anti-Lub
 Immune Ab
 Most are IgG (some are IgM)

 Reactive at 37 deg C and AHG phase

 Made in response to pregnancy or

transfusion
 Implicated with shortened survival of
transfused cells and post-transfusion
jaundice
 OPTIMAL
BLOOD ANTIBODY REACTION
REACTION ENZYME TX
GROUP CLASS PHASE
TEMP

Kell IgG 37 degC AHG No effect

Destroys Fya
Duffy IgG 37 degC AHG
and Fyb

Enhanced
Kidd IgG 37 degC AHG
aggln

Lua IgM Lua - 4 degC Lua - RT

Lutheran Variable effect
Lub IgG Lub - 37 degC Lub - AHG
 OPTIMAL
BLOOD ANTIBODY REACTION ENZYME
CLASS REACTION PHASE
GROUP TX
TEMP
Often
4degC, RT, 37 Enhanced
Lewis IgM
sometimes degC, AHG aggln
37degC
IS and
Enhanced
I IgM 4 degC occasionally
aggln
37degC
IgM (anti- IS, 37degC, Enhanced
P 4 degC
P1) AHG aggln
 OPTIMAL
BLOOD ANTIBODY REACTION ENZYM
REACTION
GROUP CLASS PHASE E TX
TEMP
MNS 4 degC or IS, 37 Destroys
IgM
MN 37 degC degC, AHG Antigens
Variable
Ss IgG 37 degC AHG
effect
MINOR BLOOD GROUPS
DIEGO (ISBT 010)
 Dia served as a useful tool in anthropologic studies
of Mongoloid ancestry, Asian people, and native
South American Indians
 Associated with
-Southeast Asian Ovalocytosis,
-Congenital Acanthocytosis,
-Hereditary Spherocytosis
 Dia and Dib antigens: located on the anion
exchange molecule (AE-1)
 an intergral transport protein involved in anion
exchange of bicarbonate for chloride in the RBC
membrane
Diego (ISBT 010) cont...
 Two sets of independent pairs of antithetical
Ags: Dia/Dib and Wra/Wrb and three-low
incidence Ags: Wda, Rba, and WARR
 Wright Ags were first classified as an
independent BGS an later as a collection
 Both anti-Dia and anti-Dib are characterized as
red-cell stimulated, IgG Abs that do not bind
the C' and reactive in indirect Antiglobulin test
 Two type of anti-Wra have been observed: a
naturally looking IgM and immune-stimulated
IgG
CARTWRIGHT (ISBT 011)
 Composed of two antigens: Yta, a high incidence
Ag present in 99.8% of general population and
antithetical partner, Ytb, a low incidence Ag
presnt in 8% of the general population
 Yt antigens have been located on erythrocyte
acetylcholinesterase, an enzyme involved in
neurotransmission
 Both anti-Yta and anti-Ytb are IgG antibodies
reactive in indirect antiglobulin test and are
considered to be clinically important because
they are predominantly red cell stimulated
XG (ISBT 012)
 Discovered in 1962
 Encoded by the short arm of the X
chromosome
 Named in association with the X
chromosome and Grand Rapids, Michigan,
the home of the first antibody producer
 Antigens: Xga, CD99

 May bind the complement

 No reported case of HTR or HDN

SCIANNA (ISBT 013)
 SC system is composed of the Sc1, Sc2, and
Sc3 antigens
 Most anti-Sc1 and anti-Sc2 antibodies are IgG,
red cell stimulated, and react in indirect
antiglobulin test, neither anti-Sc1 nor anti-Sc2
has been implicated in transfusion reactions
 Anti-Sc3 is characterized as IgG, red cell
stimulated, and reacting in IAT phase of testing,
it has been linked to mild transfusion reactions
DOMBROCK (ISBT 014)
 Doa, Dob
 Gregory(Gya), Holley (Hy), and Joseph (Joa)

 Anti-Doa and anti-Dob are described as IgG, red

cell stimulated antibodies that react primarily in
IAT with polyethylene glycol (PEG) or enz
enhancement, neither anti-Doa nor anti-Dob
has been assoc. with clinical HDN; however
both has been reported to cause acute to
delayed transfusion reactions
COLTON (ISBT 015)
 Composed of three Ags: Coa, Cob, and Co3
(formerly Coab)
 CO antigens have been located on the
transport protein known as channel-forming
integral protein (CHIP), which forms the primary
eythrocyte water channel and is responsible for
water permeability.
 CHIP and CO antigens are expressed in the
tissues of the proximal and descending tubules
and the collecting ducts of the kidney and are
believed to account for 80% of reabsorption of
water.
Colton (ISBT 015) cont...
 Both anti-Coa and anti-Cob are IgG, red
cell-stimulated and reactive in indirect
atiglobulin tetsing, and assoc. with acute to
delayed transfusion reactions.
 Anti-Co3 is characterized as IgG, red cell-
stimulated, complement binding, and
reacting in the IAT, antibody has been
assoc. with causing severe HDN.
LANDSTEINER-WIENER
(ISBT 016)
 Discovered in 1940 by Lansteiner and Wiener
 Named after the Rhesus monkey

 Antigens: Lwa (high incidence), Lwb (low

incidence)
 Enz treatment: variable effects (sensitive to
pronase and DTT)
 Do not bind complement

 Causes mild HTR and HDN

Landsteiner-Wiener (ISBT 016) cont...
 Thought to be the same as human-derived Rg
Ags and Abs but later found to be different
 Rhnull rbcs types as Lw (a-b-)

 Reacts with D+ cells

 Differentiated from Rh by DTT

DTT + Rh = REACTION
DTT + LW = NO REACTION
CHIDO/RODGERS (ISBT 017)
 Includes nine antigens which are subvdivided
into six Ch antigens, two Rodgers antigen, and
the WH antigen
 It was postulated that CH/RG antigens were
assoc. with the human leukocyte antigen (HLA)
system
 Alleles for RG and CH have been located on
two closely linked genes known as C4A and
C4B on chromosome 6
 Antigen products have been demonstrated on
the C4d fragments of the C4A (Rodgers) and
C4B (Chido) glycoproteins of the C4
complement component.
Chido/Rodgers (ISBT 017) cont...

 Formerly, the anti-Ch/Rg antibodies were

collectively grouped as HIGH TITER, LOW AVIDITY
(HTLA)
Note:
High Titer - 1:64 or greater
Low Avidity - 1+ or less through all titer dilutions
 If you suspect HTLA Abs:
1. Titer px serum; HTLA Abs will titer out a long way
(1:64) while others will not, then you can...
2. Neutralize px serum with normal fresh plasma, it
will remove the HTLA Ab.
Note:
 Otherantibodies formerly classified as
HTLAs owing to similar serologoc
reactivity:
Anti-Kn (Knops)
Anti-JMH (John Milton Hagen)
Anti-Yka (York)
Anti-Csa (Cost)
Anti-McC (McCoy)
Chido/Rodgers (ISBT 017) cont...
 Anti-Ch/Rg antibodies are usually
stimulated in multi-transfused individuals
lacking one or more of the corresponding
antigens, these antibodies are IgG and
weakly reactive in the indirect antiglobulin
phase of testing.

 Note:

Antigens are NOT INTRINSIC to the red cell

but are adsorbed in the plasma.
GERBICH (ISBT 020)
 Complex system that is composed of three high-
incidence antigens (Ge2, Ge3, Ge4) and four
low-incidence antigens (Wb, Lsa, Ana, and Dha)
 Inherited on chromosome 2 and are expressed
on glycophorins C and D (GPC and GPD)
 Gerbich-negative phenotypes are very rare
outside Papua, New Guinea
 Individuals of the Leach phenotype (GE:-2, -3, -4)
present with a change in erythrocyte morphology
in the form of elliptocytosis
Gerbich (ISBT 020) cont...
 Anti-Ge2 and anti-Ge3 have also been noted
as naturally occurring IgM immunoglobulins
and as autoantibodies, implicated in causing
acute to delayed transfusion reactions but not
in cases of clinical HDN
 Antibodies to the low-incidence GE antigens;
anti-Wb, anti-Lsa, anti-Ana, and anti-Dna are
described as predominantly IgG with an IgM
component and unable to bind complement
CROMER (ISBT 021)
 Composed of seven high-incidence Ags (Cra,
Tca, Dra, Esa, IFC, UMC and WESb) and three
low-incidence Ags (Tcb, Tcc, and WESa)
 Antigens are carried by delay accelerating
factor (DAF), involved with the regulation of
complement activation by accelerating the
decay of C3 and C5 convertase
 Anti-CROM antibodies are described as
predominantly IgG1, red cell-stimulated and
reactive in indirect antiglobulin testing
 CROM antibodies are rarely observed, majority
of examples seen in blacks
KNOPS (ISBT 022)
 Composed of five Ags: Kna (Knops), Knb,
McCa(McCoy), Sla, and Yka (York)
 Alleles for KN blood group have been located
on chromosome 1 with antigens residing on
complement receptor 1 (CR1)
 KN antibodies are characterized as IgG, red
cell-stimulated and reacting weakly and
variably in the indirect antiglobulin phase
 Abs are of little clinical importance because
none has been assoc. with causing clinical
HDN or HTR
INDIAN (ISBT 023)
 Composed of two antithetical antigens, Ina
and Inb which are relatively low and high
incidence, respectively
 IN antigens are carried on hematopoietic
isoform of the CD44 marker, which is known for
its immune adhesion properties
 IN antibodies are characterized as IgG, red
cell-stimulated, and reactive in indirect
antiglobulin testing
 Causes decreased rbc survival
OK (ISBT 024)
 Discovered in 1979 from a Japanese
woman named Mrs. Kobutso, established
in 1999
 Antigen: Oka

 Do not bind the complement

 Causes decreased rbc survival but not

HDN
 Only one anti-Oka known
RAPH (ISBT 025)
 Discovered in 1999 and named after first
antibody producer
 Antigen: MER2 (Monoclonal Eleanor
Roosevelt)
 Usually binds complement (2/3)

 No reported case of HDN

 One patient showed signs of HTR but not

definitive
JOHN MILTON HAGEN
(ISBT 026)
 Formerly belonging to ISBT 901 007
 Given its own system when gene loaction
was found in 2004
 Antigen: JMH 1-6

 No reported case of HTR and HDN

 Antigen expression decreases with age

 JMH Ab common in early patient with

acquired JMH status
GIL (ISBT 029)
 Antigen: GIL
 Antigens carried in the glycerol transporter
acquaporin 3 (AQP3), an intrinsic protein
 GILnull phenotype caused by a frameshift
and premature stop codon
RH-ASSOCIATED GLYCOPROTEIN
(RHAG) (ISBT 030)
 Antigen: Duclos (previously 901 013) and
OIa / RHAG2 (previously 700 043)
 Antibodies are usually IAT-reactive IgG
 RhAG is glycosylated on the first
extracellular loop
ISBT SYSTEM SYMBO GENE CHROMOSOMAL
CD NO.
NO. NAME L NAME LOCATION

031 FORS FORS GBGT1 9q34.13-q34.3

032 JR JR ABCG2 4q22.1 CD338

033 LAN LAN ABCB6 2q36

034 VEL VEL SMIM1 1p36.32

035 CD59 CD59 CD59 11p13 CD59

036 Augustine AUG SLC29A1 1 6p21.1

JR (ISBT 032)
 Formerly ISBT 901 014
 Jra is a high incidence Ag

 Jr (a-) phenotype is common among

Japanese
 Fully developed at birth

 RBC immune IgG, may bind complement

 Rarely causes HTR and HDFN (1 case)

 Resistant to ficin, papain, DTT and glycine-

EDTA
LAN (ISBT 033)
 Langereis

 FormerlyISBT 901 002

 May bind the complement

 Reacts with AHG

 Causes mild HDN and severe HTR (rare)

Provisional Blood Group Systems
 ISBT
034: Vel Blood group system
(VEL), formerly ISBT 212

 ISBT
035: CD59 Blood group system
(CD59)

 ISBT036: Augustine Blood group

system (AUG), formerly ISBT 901003
VEL (ISBT 034)
 Formerly ISBT 212
 RBC immune

 HTR (rare) ; no HDN due to weak expression on

cord cells
 In vivo and in vitro hemolysis demonstrated

 In serum, usually demonstrates in vitro

hemolysis
AUGUSTINE (ISBT 036)
 Ata

 Formerly ISBT 901 003

 RBC Immune: IgG

 Reacts in AHG

 At (a-) present only in BLACKS as well as

anti-Ata
 Causes moderate HTR, no HDN
OTHER BLOOD GROUP
COLLECTIONS
 ISBT 205 - Cost
-Antigens: Csa (high frequency), Csb (low
frequency)
-No HTR or HDN
 ISBT 207 - Ii

-Antigen: I (refer to ISBT 207)

 ISBT 208 - Er

-Antigens: Era (high frequency), Erb (low

frequency)
-No HTR or HDN
OTHER BLOOD GROUP COLLECTIONS
 ISBT 209 - GLOB
-Antigens: PX2, LKE (formerly under ISBT
003)
 ISBT 210 - Lewis

-Antigens: Lec, Led (formerly under ISBT

007)
Sda [Sid] (ISBT 901 012)
 RBC immune: IgM with a few IgG
 RT and AHG reaction enhanced with enzymes

 Mixed field agglutinates, shiny refractile

agglutination
 Decreased expression in pregnancy and cord
cells
 No HDN and HTR is very rare

 Antibodies are neutralized with urine from Sd

(a+) individuals
HUMAN LEUKOCYTE ANTIGENS (HLA)
DETECTABLE ON RBCs
 Bga antigen corresponds with HLA-B7
 Bgb antigen with HLA-B17

 Bgc antigen with HLA-A28

 Bg antibodies are IgG and acting weakly abnd

variably in IAT
 Not implicated on HDN and HTR

 Antibodies are usually contaminant of

polyclonal typing sera
 Destroyed by EDTA-glycine-HCl and/or
chloroquine
 Adsorbed by human platelet concentrate
HIGH INCIDENCE ANTIGENS UNRELATED TO
THE PRINCIPAL BGS

 Public Antigen
 High frequency occurring in 99.9% of
population
 Examples: Augustine (Ata), Cromer (Cra),
Ena, Gerbich (Ge), Gregory (Gya), Holley
(Hy), Jacobs (Jra), Joseph (Joa), Langereis
(Lan), Oka, and Vel (Ve)
 At (a-), Cr (a-),a nd Jo (a-) phenotype are
predominantly found in Blacks
LOW INCIDENCE ANTIGENS UNRELATED TO THE
PRINCIPAL BGS

 PrivateAntigen
 Has an incidence of <1%

 Examples include the Wright blood group

systems, which is independent of all other
group systems, as well as Swann (Swa), By,
Mta, and Tra antigens
REMEMBER! :)

 I. Neutralization Technique
a. Anti-P1 : hydatid cyst fluid/pigeon
droppings/turtle dove egg white
b. Anti-Le : secretor saliva / pooled plasma or
serum
c. Anti-Ch/Rg : pooled plasma / serum
d. Anti-Crom : conc. seum, plasma or urine
e. Anti-Sda : Guinea pig urine, urine of Sd(a+)
individual
f. Anti-I : mother's milk, human breast milk
g. Anti-H : saliva
 Decreased RBC survival
REMEMBER! :) McLeod, Indian, Ok

 II.Enzyme Technique
a. Enhanced:
Kidd, Rh, Lewis, I, P (P1)
b. Inactivated:
MNSs, Fy, Xg, Yt, Ch/Rg

 III.
Chemicals
ZZAP (cysteine activated papain)
- contains Dithiothreitol and papain