BLOOD BANK DONOR CENTER

 Involve in promoting voluntary blood donation Parent A B O

from the evaluation of potential donors to the Alleles
collection of both in house and mobile blood A AA AB AO
donation procedures (A) (AB) (A)
 The section is manned by nurses skilled in B AB BB BO
advocacy, donor recruitment (AB) (B) (B)
 Genotype- inheritance O AO BO OO
 Phenotype- manifested (A) (B) (O)
 O- recessive
Formation of A, B, H Antigens
CONTENT GUIDELINE  ABO genes code not for the antigen themselves
1. ABO and Rh but for the production of glycosyltransferase
2. Antibody screening and ID that add immunodominant sugar to a basic
a. Duffy, Il Kell, Kidd, Lewis, MNS, P Rh, precursor substance
Multiple antibodies GENE Glycosyltransferase Immuno
3. Crossmatch and Special tests dominant Sugar
a. DAT, Phenotyping, Genotyping, Elution/ H gene L- fucosyltansferase L- fucose
adsorption, Ab titer, Prewarm A gene N- acetylgalactosaminyl N- acetyl-D-
technique, Rosette and Kleihauer Betke transferase galactosamine
4. Blood donation. Transfusion reactions and B gene D- galactosyltransferase D- galactose
Hemolytic disease of the newborn
a. Donor request, testing Biochemical Structure of ABO
b. Transfusion therapy- RBC, PLT, FFT,  On the 37th day of fetal life attachment of
CRYO, RhIg immunodominant sugar occurs on the RBC
membrane and it is dependent on ABH genes
ABO inherited.
 Most transfusion associated facilities due to AA  N- acetylgalactosaminyl
ABO incompatibility H structure: AO transferases
 Described by Karl Lansteiner (1901) BB  Galactosyl
 Most significant Transfusion practice H structure: BO transferases
 Transfusion of ABO incompatibility may cause
severe intravascular hemolysis or Acute
hemolytic transfusion reaction

ANTIGEN INHERITANCE
 ABH gene located at Chromosome 9
 ABH genes are inherited in a codominant
manner following simple Mendelian genetics
law GALNAC- N- acetylgalactosaminyl immunodominant
Phenotype Genotype sugar responsible for A specificity
O OO D- Galactose- immunodominant sugar responsible for B
A AA, AO specificity
B BB, BO
AB AB
ABO ANTIBODIES Precursor Type 2 Type 1 and Type 2
 Are mostly naturally occurring antibodies that chain
are detectable 3- 6 months after birth following Linkage 14 linkage 1 3 linkage
exposure to ABO like antigens in the Regulating Zz gene Sese gene
environment. gene
 Are mostly IgM and react best at RT A SUBGROUPS
GROUP ANTIGEN ANTIBODY A SUBGROUP
O H Anti A, B, AB GROUP FREQUENCY ANTI ANTI ANTI ANTI
A A, H Anti B A B A1 H
B B, H Anti A A1 80% ++++ - ++ -
AB A, B,H None A2 < 20% +++ - - ++
 A2 cells- has more antigen sites for H antigen
INCIDENCE  A1 cells- H antigen site occupied by both A and
GROUP WHITES BLACKS A1 antigen
O 45% 49% B SUBGROUP – are in frequent
A 40% 27%  Anti B Lectin- Bandeirea Simplicifolia
B 11% 20%  Bombay (Oh) Phenotype
AB 4% 4% - the allele of h are very rare and does not
produce the L- fucose necessary for the
BT Forward Typing Reverse formation of H- structure
Typing - the genotype hh or Hnull is known as the
Anti Anti Anti A B O Bombay phenotype (Oh)
A B AB Cells Cells Cells GENERAL CHARACTERISTIC OF OH
O - - - + + -  Absence of H, A and B antigen
A + - + - + -  Presence of Anti H, Anti A and Anti B in serum
B - + + + - -  Strong reactivity with Anti I reagent
AB + + + - - -  A recessive inheritance
 No reaction with Anti H lectin (Ulex europaeus)
Interaction Of Sese, Zz And ABH Genes REACTIVITY OF ANTI H ANTISERA ON ANTI H LECTIN
 Sese System- regulates the formation of H WITH ABO BLOOD GROUPS
antigen and subsequently of A and B antigen is  Greatest amount of H Ag: O>A2>B
secretory cells.  Least amount of H Ag: A2B>A1>A1B
 Zz System- regulates production of H antigen on
erythrocyte ABO DISCREPANCIES
ABH Antigen ABH Soluble  Exist when the result of RBC test (Antigen) don’t
Substance agree with the serum test (Antibody).
Found where RBC, In all body  When discrepancy is encountered:
Epithelium secretion  Results must be recorded
tissue, BM,  Interpretation of the ABO group must
Cells be delayed until the discrepancy resolve
Secreted Glycolipids Glycoproteins GENERAL RULES TO RESOLVE DISCREPANCY
substance 1. Always re- test
1st sugar in Glucose N- 2. Check for clinical/ technical errors
precursor acetylgalactosamin a. Inadequate identification of blood
substance samples, test tube/ slide
 b. Cells suspension either too heavy or too b. Increase permeability of
light intestinal wall causes
c. Clinical errors adsorption of B-like bacterial
d. Mix up samples polysaccharide onto red cells of
e. Failure to add reagent or fail to follow A or B patients (P. vulgaris/
up manufactures instruction E.coli)
f. Uncalibrated centrifuge  Antibody to low incidence Antigen (may
3. Weakest reaction is usually the one in doubt be present in reagent antisera)
4. Check results of screening cells  Excess blood group specific soluble
5. Check patient age substances (BGSS) in cases of stomach
6. Check diagnosis or pancreas
7. Check transfusion history EXCESS BGSS
GROUP 1 Anti Anti Anti A1 B O Auto
 Due to weakly reacting or missing antibodies, A B A,B Cells Cells Cells Control
most common encountered H is found in the - - - - 4+ - -
following: After washing. . .
 Newborn Anti Anti Anti A1 B O Auto
 Elderly patient A B A,B Cells Cells Cells Control
 Leukemic patients demonstrating 4+ - 4+ - 4+ - -
lyphogammaglobulinemia GROUP 3
 Patients with lymphoma  Caused by CHON or abnormalities resulting to
 Patients with congenital α gamma roleaux formation H is found in:
globulinemia  Increase globulin (Multiple myeloma,
 Patient using immunosuppressed drugs Waldenstrom, macroglobulinemia)
 Patients with immunodeficiency  Increase fibrinogenpresence of plasma
 Patient who undergo bone marrow expanders (dextran)
transplant who develop  Wharton’s jelly (+ on Cord bloods)
hypogammaglobulinemia from therapy Anti Anti Anti A1 B O Auto
 Artificial Chimera A B A,B Cells Cells Cells Control
- blood transfusion 4+ 2+ 4+ 2+ 4+ 2+ 2+
- bone marrow transplant Probably type: A
- exchange transfusion GROUP 4
- fetal maternal bleeding  Due to miscellaneous problems seen in:
GROUP 2  Polyagglutination (Hubener- Thomson
 Due to missing antigen and is the least Friedenreich Phenomenon)
frequently encountered. It is found in: - Spontaneous red cell agglutination
 Subgroup of A or B by most normal human serum
 Leukemia may yield weakened A or B - Occurs to exposure of a hidden rbc
antigen antigen (T antigen) in patients w/
 Hodgkin’s disease bacterial and viral infections
 Acquired B  Cold reacting antibody (Allo, Auto)
a. When bacterial enzyme  Unexpected ABO agglutinins
(Proteus vulgaris) modify N-  Warm antibodies
acetylgalactosamine into D-  RBC with antibody phenotype
galactose) – Most Common  Antibodies other than anti A and anti B may
react to form antigen antibody complexes
 Anti Anti Anti A1 B O Auto o Perform cold antibodies panel (Prewarm,
A B A,B Cells Cells Cells Control adsorpotion, treatment with sulfhydryl
1 - - - - - - - compounds)
Possible cause: Group O newborn or elderly patient may
have α gammaglobulinemia/ may be on RH SYSTEM
immunosuppressed drugs  Described by Levine and Stetson (1939)
Resolution: Enhance reverse grouping  Most clinically important system after ABO
 Formation of Anti- D always results from
Anti Anti Anti A1 B O Auto transfusion or pregnancy
A B A,B Cells Cells Cells Control  High immunogenicity more than 80% of D-
2 4+ - 4+ 1+ 4+ - - negative persons who receive a D- positive
Possible cause: Subgroup of A probable A2 indvidual transfusion are expected to develop Anti- D
with Anti- A1  Rh is based from Ab produced by guinea pigs
Resolution: Use Anti- A1 lectin and rabbits when transfused with rhesus RBC;
Anti Anti Anti A1 B O Auto and this Abs agglutinate 85% of human cells
A B A,B Cells Cells Cells Control
3 - - - 4+ 4+ 4+ - NOMENCLATURE/ TERMINOLOGIES
Possible cause: Bombay phenotype Fisher – Race/ DCE Terminology (England)
Resolution: Test with Anti- H lectin  Rh antigens are produced by three closely
linked sets of alleles. This five major antigens
Anti Anti Anti A1 B O Auto defined are D, C, E, c and e
A B A,B Cells Cells Cells Control D gene D antigen
1 4+ 1+ 4+ - 4+ - - 3 distinct
C gene C antigen
antigens on
Possible cause: E gene E antigen
red cell
o Group A with acquired B antigen
membrane
o Low incidence antibodies in the reagent
antisera (Anti- B reagent) Wiener/ Rh- hr Terminology (United States)
Resolution:  Rh antigen has series of blood factors, in which
o Check history of patient for lower GI CHON or each factor is an antigen recognized by an
septicemia. Acidify Anti- B typing reagent to pH antibody
6.0 by adding 1-2 drops of 1N HCl to 1ml of  Example: Antigen –RhO
Anti- B antisera Blood factors –RhO, hr’, hr”
o Use another reagent antisera with different lot Rosenfield/ Alpha numeric Terminology
number  It has no genetic basis and is based according to
the order of discovery
Anti Anti Anti A1 B O Auto ISBT (International Society of Blood Transfusion)
A B A,B Cells Cells Cells Control  It has also no genetic basis
4+ 4+ 4+ 2+ 2+ 2+ 2+
Possible cause: Wiener Fisher- Race Rosenfield
o Roleaux (Multiple myeloma) Rho D Rh1
o Cold autoantibody (Probable bloodtype: AB Rh’ C Rh2
with Auto anti) rh” E Rh3
Resolution: hr’ c Rh4
o Wash red cell hr” e Rh5
LW- is the antigen closely associated phenotypically
with Rh
 - It is formerly known as Rh25 - + Rh Positive
Ne- a- is the recently discovere anti- thetical antigen to - - Rh Negative
LW
Rh locus- located at chromosome 1 IMMUNOGENICITY OF COMPOUND Rh ANTIGENS
Shorthand Wiener Blood Fisher- D > C > E > C > e
factors Race G ANTIGENS
R0 Rh0 Rho, hr’,hr” Dce  Are produced by same Rh gene complexes that
R1 Rh1 Rho, rh’,hr” DCe produce C and D antigens. Most C-positive and
R2 Rh2 Rho, hr’,rh” DcE D-negetive RBC’s are also G-positve
Rz Rh2 Rho, rh’,rh” DCE F(ce)
r rh hr’, hr” dce  a compound antigen of c and e
r’ rh’ rh’, hr” dCe Rh;(cCe)
r” rh” hr”, rh” dcE  a compound antigen of C and e
rγ rhγ rh’, rh” dCE Rh ANTIBODIES
 Are usually igG1 or igG3 rbc—stimulated, either
R1 or DCe/dce- most common in whites approximately during transfusion of during pregnancy
35%  Do not agglutinate in saline.
RCr or Dce/dce- most common in blacks approximately  React best at 37’C and can be demonstrated by
42% testing in high protein media or by the indirect
D ANTIGENS antiglobulin test/
 Characterized as non- glycosylated CHON on red  Reactions is enhanced by the use of enzyme
cell membrane D antigen treated rbc’s
 The only Rh antigen that undergoes routine  Do not bind complement
testing except in the case of investigation of  They cross the placenta and can cause HDN
unexpected antibodies SYNDROME
Variation of the Rho (D) Antigen  It expresses no Rh antigen o red cell and the
 Du phenotype- weakened expression of the D phenotype is expressed as ---|---
antigen  The following are symptoms:
MECHANISM FOR DU PHENOTYPE OCCURRENCE  Stomatocytosis
 Genetic weak D/ Genetics Du/ Low grade Du  Reticulocytosis
It is an inherited weakened expression of D antigen  Compensated hemolytic anemia
in this mechanism, antigen is complete but few in  Increase HbF bilirubin
number  Decrease haptoglobulin, DFT
 C trans/ Position effect/ Gene interaction  Slightly decrease Hb, Hct
effect Anti- Rh: 29
The allele carrying D is trans to an allele carrying C  The antibody of broadest specificity
For example: DCe/dCe or Dce/dCe  Described as anti total Rh
 D Mosaic antigen  Can be produced by Rh null individual
There is missing components of D antigen subunits FALSE REACTION OF RH TYPING
The Anti D formed by D mosaic individual can cause False Positive Reaction
HDN and HTR  Positive DAT- most common cause of Rh- typing
(Du typing) discrepancies
DETERMINATION OF D STATUS
 Roleaux
Anti- D Indirect AHG Interpretation
 Cold agglutinins- patient sample should be
+ Rh Positive
warmed to 37C and immediately re tested
False Negative Reaction may cause by LEWIS ANTIGEN
 Incorrect cell suspension (cell suspension too
Galactose
heavy or too light)
N-acetyl
POST ZONE EFFECT- antigen excess glucosamine
Le galactose
PROZONE EFFECT- antibody excess L- Le b
gene
 Improper procedure FUCOSE N- acetyl structure
adds
galactosamine
The following reactions were obtained
Red cell
Cells tested with Serum tested with membrane
Anti- A = 4+ A1 cells = 2+  Lewis substance (in secretion) are glycoproteins
Anti- B = 3+ B cells = 4+
 Lewis antigen (cell bound Ags) are
Anti- A,B = 4+
glycophospholipids
The technologist washed the patient cells with saline  Poorly developed at birth (not found in
and added two drops of saline to the reverse grouping. newborn RBC)
Upon repeat testing, the following results were   Le (a-b) Le (a+b-) Le (a+b+) Le (a+b)
obtained Le (a-b+) the tru phenotype
Cells tested with Serum tested with INHERITANCE
Anti- A = 4+ A1 cells = 0  Le Genes
Anti- B = 0 B cells = 4+  Located on the short arm of Chromosome
Anti- A,B = 4+ 19
The results are consistent with:  Genes does not actually code for the
a. Acquired immunodeficiency disease production of Lewis antigen but,
b. Bruton’s agammaglobulinemia  Rather, produces a specific L-
c. Multiple myeloma fucosyltransferase to type 1= precursor
d. Acquired “B” Antigen substance
Refer to the data:  Lewis Phenotype
ANTISERA REACTION  Le (a+b-): Non secretors
Anti- D +  All Le (a+b-) are non-secretors of ABH
Anti- C - substance
Anti- E -  Lea substances is secreted regardless of
Anti- c + secretor status
Anti- e +
Which of the following is the possible genotype? N- acetyl glucosamine
a. R1r’ galactose
b. R1Ro Le gene L- Le a
N- acetylgalactosamine
adds FUCOSE Structure
c. Ror Red cell membrane
d. rr (CHON Backbone)

LEWIS BLOOD GROUP SYSTEM

 Lewis antigen are manufactured by tissue cells
and secreted into the body fluids the adsorbed
onto the red cell membrane (not really an
integral part of the red cell membrane)
  Are found on red cells, not in body fluids and
Galactose secretions
N- acetyl glucosamine  U- stands for universal RBCs with the S or s
H gene L- galactose H type 1 antigen also have the V antigen
adds FUCOSE N- acetylgalactosamine Structure
INCIDENCE OF PHENOTYPES
Red cell membrane MN: 50% S: 45%
(CHON Backbone)
S: 55% U+: Common among whites
V: Common among N: 20%
blacks
 Le (a-b+): Secretors
M: 30% ENa: >99.9 %
 This phenotype is the result of the
genetic interaction of Le and Sese genes
MNSS ANTIBODIES
 Both Lea- soluble and Leb- soluble
 Anti M and Anti N
antigens can be found in the secretion
 Mostly IgM, naturally occurring cold
but only Leb adsorbs onto the red cell
reactive saline agglutinins that don’t bind
membrane
complement or react with enzyme treated
H gene adds L- FUCOSE Galactose cells (Destroyed enzymes)
 Anti N seen in renal patients who are
 Le (a-b-): Secretors or Non secretors dialyzed with equipment sterilized with
 80% ABH Secretors formaldehyde
 20% ABH Non secretors  Anti M reaction enhanced by acidification
LEWIS ANTIBODY  Anti S and Anti S
 Are usually naturally occurring IgM; react best  Most are IgG reactive at 37C and the
at RT or lower; considered clinically insignificant antiglobulin phase
 Binds complement and therefore capable of  May bind complement and have been
triggering in vitro hemolysis associated with HDN and HTRs
 Enhanced by enzyme treatment
 Readily neutralized by Lewis blood group P BLOOD GROUP SYSTEM
substances PHENOTYPES DETECTABLE POSSIBLE Ags
 Anti Lea- most commonly encountered Ab Ags
of the Lewis system P1 P, P1 -
Biologic Significance P2 P Anti- P2
 Leb has receptors for Helicobacter pylori P - Anti- PP2 PK
 Lex antigen is marker for Reed- Sternberg of P1K P2, PK Anti- P
K K
Hodgkin’s Disease Pe P Anti- P , Anti- P1
P ANTIGEN
OTHER BLOOD GROUP SYSTEM  Consist of 3 antigens P, P1 and PK which are
MNSs U BLOOD GROUP SYSTEM biochemically related to the CHO chain that
 the two loci system makes up the ABH and I antigen
MNSS U ANTIGENS  Located at Chromosome 22
 MNS are inherited as close linkage, MN is  P1 antigen are poorly developed at birth
associated with glycophonin A: Ss is associated P ANTIBODIES
with glycophonin B  Anti P1
 Located at Chromosome 4  Naturally occurring IgM in the sera of P2
 MNS antigens are important markers in individual weak cold reactive saline
paternity testing agglutinin
  Can be neutralized with soluble P1
substance in hydatid cyst fluid
(Echinococcus granulosus infen)
 Anti P
 Naturally occurring alloantibody in the sera
of all PK individuals
 AUTOANI-P (Donath Landsteiner Antibody)-
IgG biphasic hemolysis (attaches to rbcs in
cold, lyses rbc in warm temperature)
associated with Paroxysmal Cold
Hemoglobinuria
 Anti PP1 PK (Anti Tja)
 Predominantly IgM, binds complement
 Associated with spontaneous abortion in
early pregnancy
 May demonstrate invitro hemolysis