OTHER BLOOD GROUP

SYSTEM
Prepared by: Mark Christian D.L. De La Cruz, RMT
LEWIS BLOOD GROUP SYSTEM
• ISBT 007
• Lewis gene (Le) codes for the production of fucosyltransferase
enzyme
• This gene codes for a specific glycosyltransferase, α-4-
Lfucosyltransferase, which transfers L-fucose to type 1 chain (type
1 precursor) oligosaccharide on glycoprotein or glycolipid
structures.
• Erythrocytes acquire the Lewis phenotype by adsorbing Lewis
substances from the plasma, rather than membrane bound antigens
Precursor molecule
• Type 1 precursor chain
• beta 1-3 linkage
• Type 2 precursor chain
• beta 1-4 linkage
• The inheritance of the Le gene acts in
competition with ABO genes, adding L-fucose
to the GlcNAc sugar of the common precursor
structure manufactured by tissue cells.
• It is then secreted and adsorbed onto the
RBCs, lymphocytes, and platelet membranes
from plasma.
• Lewis antigen is also found on other tissues,
such as the pancreas, stomach, intestine,
skeletal muscle, renal cortex, and adrenal
glands.
Lewis Phenotypes
• Le (a+ b-) phenotype:
• Lea substance is secreted regardless of the secretor status.
• All Le (a+ b-) individuals are nonsecretors of ABH substances.
• Le (a- b+) phenotype:
• The genetically independent Sese, ABO, Hh, and Lewis genes are
intimately associated in the formation of the Leb antigen.
• The phenotype Le (a- b+) is the result of the genetic interaction of
Lele and Sese genes.
• Le (a- b-) phenotype:
• Results in point mutation of Le gene, rather than absence of the Le
gene.
• Le (a- b-)
• These mutations give rise to a nonfunctional or partially active
Lewis transferase (Lew) causing the negative expression of the
Lewis antigen on the RBCs.
• Presence of substances in secretions will depend on the secretor
status of the individual.
• The Le (a- b-) nonsecretors express type 1 precursor, whereas
the Le (a- b-) secretors express H type 1 substances plus ABH
antigens associated with the related ABO genes inherited.
 SUBSTANCE IN THE RED CELL
GENOTYPE
BODY FLUIDS PHENOTYPE
ABH, lele, sese NONE ABH Le (a-b-)
ABH, lele, SeSe or
ABH ABH Le (a-b-)
Sese
ABH, LeLe or Lele,
Lea ABH Le (a+b-)
sese
ABH, LeLe or Lele,
ABH, Lea, Leb ABH Le (a-b+)
SeSe or Sese
Development and Changes of Lewis Antigens
After Birth
• In plasma there are no detectable Lewis glycosphingolipids at birth.
• Therefore, cord blood and RBCs from newborn infants phenotype
as Le (a- b-).
• Infants that inherit LE and Se genes.

Le (a- b-) Le (a+ b-) Le (a- b+)

10 days 6-7 yrs
• Infants that inherit the Le and se genes.
Le (a- b-) Le (a+ b-)
10 days
DURING PREGANCY
• A decrease in expression of Lewis antigens has been demonstrated
on RBCs from many pregnant women, resulting in Le(a- b-)
phenotypes during gestation.
Lewis Antibodies (General aspects)
• Naturally occurring
• IgM in nature
• Complement activation (+) → can cause in vivo and in vitro hemolysis.
• Reactivity is enhanced when treated with proteolytic enzymes.
Anti-Lea
• Most commonly encountered antibody of the Lewis system.
• Produced by Le (a- b-) phenotypes
• MOSTLY IgM. Some may have IgG form but does not bind to the RBCs
as efficiently as IgM anti-Lea form.
• The Lea antibody is frequently detected with saline-suspended cells
at room temperature.
• Sometimes react at 37oC and Coombs (AHG) phase.
• Easily neutralized with plasma or saliva that contains Lea substance.
• Phenotype Le (a- b+) : do not make anti-Lea.
Anti-Leb

• Not as common and not as strong as anti-Lea.

• IgM
• Usually produced by Le (a- b-)
• Occasionally produced by .
• Neutralized by plasma or saliva containing Leb substance.
Clinical Significance of Lewis Antibodies
• Some cases of HTR caused by anti-Lea have been reported.
• Cases of in vivo RBC destruction due to anti-Leb also have been
reported.
• Generally, Lewis antibodies are considered insignificant in blood
transfusion practices.
• It can be neutralized by the Lewis substances present in the
plasma and can thereby be decreased in quantity.
• Lewis antigens dissociates from the RBCs as readily as they bind to
the RBCs.
• Lewis antibodies are generally IgM and therefore cannot cross the
placenta and cause HDN.
• Lewis antigens are not fully developed at birth
MNS Blood Group System
• Discovered by Landsteiner and Levine in 1927: M and N antigens
• Immunizing rabbits with human RBCs.
• Data from family studies suggested that M and N were antithetical
antigens.
• In 1947: Walsh and Montgomery discovered S antigen that appeared
genetically linked to M and N.
• Antithetical partner of S is the s antigen. (discovered in 1951).
• 1953: Wiener discovered U antigen.
M and N antigen
• Found on a well-characterized glycoprotein known as .
• major RBC sialic acid–rich glycoprotein.
• Well developed at birth
• Found on RBCs BUT NOT in platelets, lymphocytes, monocytes and
granulocytes.
• Easily destroyed or removed/inactivated by proteolytic enzymes.
• Because M and N are located at the outer end of GPA.
• Antigens M and N differ on the 1st and 5th amino acids.
• Shows dosage in serologic reactions
S and s antigens
• Located on smaller glycoprotein called .
• S and s are differentiated by the amino acids at position 29 on GPB.
• S has _______________ whereas s has _____________
• Well developed at birth
• Destroyed or removed/inactivated by proteolytic enzymes.
• Except for Trypsin→ does not destroy S and s antigens.
• Found on RBCs BUT NOT in platelets, lymphocytes, monocytes and
granulocytes.
Other MNSs antigens
• U antigen:
• Located on GPB
• found on RBCs of all individuals except about 1 percent of American blacks
(and 1 to 35 percent of African blacks)
• U antigen negative cells are also S-s-(Ss Null)
• En (a-) antigen:
• High Frequency antigen
• Failure to produce Glycophorin A results in Ena negative red cells
MNSs antibodies
• Anti-M:
• Naturally occurring
• IgM or IgG
• Do not bind complement: regardless of their Ig class.
• Do not react with enzyme treated RBCs.
• Demonstrate dosage
• Reacts better with M+ N- than M+ N+ RBCs.
• Enhanced reaction at pH 6.5.
• Some react only with red cells exposed to glucose solution
• Not clinically significant unless it reacted at 37oC or AHG phase.
• Rarely causes HDN.
• Anti-N:
• Naturally occurring
• IgM or IgG
• Do not bind complement: regardless of their Ig class.
• Do not react with enzyme treated RBCs.
• Demonstrate dosage
• Reacts better with M- N+ than M+ N+ RBCs.
• Rarely reacts with red cells exposed to glucose solution
• Not clinically significant unless it reacted at 37oC or AHG phase.
• Rarely causes HDN.
• Anti-N reagent used in laboratory: Vicia graminea
• Anti-Nf
• seen in renal patients, regardless of their MN type, who are dialyzed on
equipment sterilized with formaldehyde.
• reacts with any N+ or N- RBC treated with formaldehyde.
• The antibody titer decreases when dialysis treatment and exposure to
formaldehyde stop.
• clinically insignificant in transfusion.
• Anti-S and anti-s:
• IgG in nature
• They may bind complement
• they have been implicated in severe hemolytic transfusion reactions with
hemoglobinuria.
• Also caused HDN
• Units selected for transfusion must be antigen-negative and crossmatch-
compatible.
• it can be difficult providing blood for a patient with anti-s.
• Anti-U
• is typically IgG and has been reported to cause hemolytic transfusion
reactions and HDN.
Anti-sera reagents
• Anti-M
• Human source
• Rabbit source
• Monoclonal source

• Anit-N
• Monoclonal
• Vicea graminea (Lectin)
P BLOOD GROUP SYSTEM
 P Blood Group Antigens
• P1, P and Pk antigens.
• Discovered by Landsteiner and Levine (1927).
• Injected human RBCs to rabbits and produced an antibody.
• They called it as .
• 1951, Levine et al.
• Describe an antibody known as anti-Tja ( now known as ).
• Made by Pnull individual.
PHENOTYPE DETECTABLE ANTIGENS POSSIBLE ANTIBODIES
P1 P & P1 NONE
P2 P Anti-P1
Anti-P and anti-P1, anti-
P null NONE
Pk
P1K PK, P1 Anti-P
P2K PK Anti-P and Anti-P1
P1 antigen
• Changes during fetal development.
• Found on fetal red cells as early as 12 weeks, but it weakens with
gestational age
• Not well developed at birth
• P1 antigen deteriorates rapidly on storage
• P1 like antigen has been found plasma and droppings of pigeons and
turtle doves, as well as in the egg white of turtledoves
• P1 in hydatid cyst fluid, extracts of Lumbricoides terrestris and Ascaris
suum
 Anti-P1
• Common, naturally occurring IgM antibody in the sera of P2
individuals
• COLD REACTIVE
• Strong anti-P1 was observed in individuals infected with Hydatid
dse.
• Not well developed at birth
• Associated with , and
infection.
• HDN is not associated with anti-P1
Anti-PP1Pk
• Originally called ___________
• Predominantly IgM, sometimes IgG
• Reacts over a wide thermal range
• efficiently bind complement, which makes them potent
hemolysins.
• Anti-PP1Pk has the potential to cause severe hemolytic transfusion
reactions and HDN.
• ASSOCIATED WITH SPONTANEOUS ABORTIONS IN EARLY PREGNANCY
• Anti-PP1Pk→ reacts with all RBCs except to the autocontrol and those
of the p phenotype.
Alloanti-P
• Naturally occurring
• Seen in sera of all Pk individuals.
• Reactivity is the same as Anti-PP1Pk
• Anti-PP1Pk→ reacts with all RBCs except to the autocontrol and
those of the p phenotype.
• Rarely seen in bloodbank (but very significant in transfusion)
• Has been associated with habitual early abortion.
Autoanti-P
• Cold-reactive IgG autoantibody
• Associated with PCH.
• The IgG autoantibody in PCH is described as a biphasic hemolysin.
• In vitro, the antibody binds to RBCs in the cold, and via
complement activation, the coated RBCs lyse as they are warmed
to 37oC
• The autoantibody typically does not react in routine test systems
but is demonstrable only by the Donath-Landsteiner test.
Donaths-Landsteiner Test
Collection of fresh blood Prepared three sets of
from the patient Test tubes
Incubation
“A” tubes: ice bath for 30 mins and 1
hour @ 37ocelsius
Incubation
“B” tubes: ice bath for 90 mins
Incubation
“C” tubes: 37o Celsius for 90 mins
CENTRIFUGED AND OBSERVED THE
SUPERNATANT FOR HEMOLYSIS
A1,B1 and C1→ add 10 drops
of patients serum, one
volume of 50% suspension of
washed Group O cells that
expressed P antigen
A2,B2 and C2→ add 10 drops
of patients serum, 10 drops
of normal serum and one
volume of 50% suspension of
washed Group O cells that
expressed P antigen
A3,B3 and C3→ add 10 drops
of normal serum, one
volume of 50% suspension of
washed Group O cells that
expressed P antigen
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 AIHA
• Anti-Pk activity can be inhibited with hydatid cyst fluid.