LECTURE 3

COMPARISON OF ABH ANNTIGENS ON

RBCS AND IN SECRETIONS
ABH Antigens A, B and H soluble
on Red Cells Substances(Secretions)
Glycolipids, glycoproteins or glycosphingolipids Secreted substances are glycoproteins
RBC antigen are only synthesized on type 2 Secreted substance are primarily on Type 1
precursor chains precursor chain
Type2 chain refers to a beta 1-4 linkage in w/c The number 1 carbon of galactose is
the number 1 carbon of the N-acetylglucosamine attached to the number 3 of the N-
sugar of the precursor substance acetylglucosamine sugar of the precursor
substance
The enzyme produced by the H gene (alpha-2-L The enzyme produced by the Se gene
fucosyltransferase) acts primarily on type 2 (alpha-2-L fucosyltransferase)
chains, w/c are prevalent on the RBC membrane preferentially acts on type 1 chains in
secretory tissues
Points Type 1 Type 2 Secretions RBCs
Linkage Beta 1-3 Beta 1-4 The Glycosphingomyelin
Origin Plasma Synthesized on oligosaccharides carrying A and B
RBC precursor are present in oligosaccharides are
soluble forms in integral part of the
Controlling H, A, B, Se H, A, and B the plasma RBC membrane,
genes and Lewis endothelial and
epithelial cells
Sources of
What are the substances responsible for A Antibodies
and B activity in secretions?
Glycoproteins that carry identical Antibodies Sources
Oligosaccharides. Anti A Fish roe and Tears
A& B Oligosaccharides that lack either Anti B Colostrum, Ascitic Fluid
proteins or lipids are found in milk and urine
KINDS OF ANTI H Anti A, B Saliva of Non-secretors,
Snails
Pts of Human Blood Plant
Differences Anti A1 D.b., Anti A absorbed
with A2 cells
Source Bombay Ulex
eurapaeus Anti H U.e., eel immunized
chicken
Activity Weaker Stronger
Use in Not used Used in CHARACTERISTIC OF ABO ANTIBODIES
Laboratory Subgrouping 1. Naturally Occuring
2. Immune Forms
Reactivity Weakly reacts Reacts with
with H Ag "O" and A2 Antibody Titer
Temperatur 37 degrees C Cold Reacting Cord Blood decreased
e Infants decresed
Adults increased
Age 5-6, 5-10 Peak
65 years old decreased

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11.4 IgG or IgM ABO ANTIGENS IN SECRETION
• Secretions include body fluids
"O" IgG/IgM like Plasma, Saliva, synovial
IgG/IgA fluid, etc
IgG/IgM/IgA • Blood group substances are
"A" IgM/IgG soluble antigens (A,B and H)
IgM/IgA • This is controlled by the H and
IgM/IgG/IgA Se genes
11.5 Strength of Reactivity SECRETOR STATUS
Anti A in O is higher than Anti A in “B” • Secretor gene consists of 2
- Type O- used as third serum in the laboratory alleles (Se and se)
- Anti A in “B” is higher than anti B in “A” • The Se gene is responsible for
- Anti A and Anti B are higher than Anti Rho or Anti D the expression of H antigen on
 Anti A – BLUE COLOR glycoprotein structure located
 Anti B - Yellow color in body secretions.

12. Other Antibody Characteristics

Points of Differences Complete (IgM) Incomplete (IgG)

Medium Saline, LISS Albumin, AHG, Enzymes
Traverses the placenta Does not Traverses
Temp. Labile Stable
Hemolysis Does not produce hemolysis Produces Hemolysis
Binding Binds complement Does not bind complement
Inactivation Inactivated Resistant
M.W. 9hun k to 1M 145 hun k - 150 hun k
Valence Bivalent Univalent
Sedimentation Constant 19S 7S
13. Presence of Complement
- C +A or B Ags+ABO Abs (act as hemolysis) • If the Se allele is inherited as SeSe
• Result: hemolysis or Sese, the person is called
because of the complement presence in the blood secretor
- Anticoagulant like Ca chelating EDTA is added - 80% of the populations are
- Abs (Calcium binding Abs can’t be detected secretorS
14. ABO system is controlled by: SECRETORS AND NON-
1. A1,A2 , B and O SECRETORS
2. H and h • Secretors express soluble forms of
3. Se and se the H antigen in secretions that can
15. ABH genes are controlled by be converted to A or B antigens
1. Se and se ( by transferases)
2. Zz • Individuals who inherit the sese
16. Sources of ABH antigens genes are called nonsecretors
1. Structural lipoproteins - These se allele is an amorph
a. Erythrocyte membrane ( nothing expressed)
b. Epithelial cells - sese individuals do not convert
2. BM antigen precursors to H antigen and
3. Kidneys has neither soluble H antigen nor
4. Lymphocytes soluble A or B antigens in body
5. Platelets Not found in granulocytes fluids.
17. Other sources
1. Pig 6. Ovarian cysts 11. Pericardial
2. UGTT 7. Seminal fluid 12. Peritoneal
3. Saliva 8. Amniotic fluid
4. Tears 9. Milk
5. Urine 10. Pleural

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SECRETORY STATUS SUMMARY SECRETOR STATUS
- The Se gene codes for the presence of the H - Inhibition/Neutralization test
antigen in secretions, therefore the presence Uses:
of A and B antigens in the secretions is 1. provides a means of assaying the
contingent on the inheritance of the Se gene relative strength or potency of water-
and H gene. soluble blood group substances.
2. Identifies the presence of ABH or Lea
substance.
• A secretor show no agglutination
• A secretor control shows no agglu.
A non secretor controls shows agglutination

ABO SUBGROUPS
ABO Group ABH • ABO subgroups differ in the amount of
Substances antigen present on the RBC membrane
- subgroups have less antigen
Secretors (SeSe or A B H • Subgroups are the results of less
Sese): effective enzymes.
A +++ 0 + • They are not as efficient in converting
B 0 +++ + H antigens to A or B antigens (fewer
antigens are present on RBC.
O 0 0 • Subgroups of A are more common than
AB +++ +++ + subgroup of B.

Non-secretors SUBGROUP OF A
(sese): • The 2 principal of subgroups of A are;
A1 and A2
A,B,O and AB 0 0 0 - Both react strongly with reagent anti A
Points of ABH in ABH in RBC - To distinguish A1 from A2 red cells, the
Diff secretions lectin Dolichos biflorus is used (Anti A1)
- 80% of group A or AB individuals are
Substance Glycoproteins Glycolipids subgroup A1
Solubility Water Soluble Fat Soluble - 20% are A2 and A2B ( fewer antigen
Biosynthes Type 1 Type 2 weaker reactions)
is A2 PHENOTYPE
Why is A2 phenotype important?
Controlling Se/se Zz - A2 and A2B individuals may produce an
genes anti A1
- This may cause discrepancies when a
THE RELATIONSHIP OF ABH cross match is done (incompatibility)
SUBSTANCES AND ABO GROUP What is the difference between the A1 and
ABH Subs. In saliva (SeSe or Sese A2 antigen?
ABO Blood Group A B H - It is quantitative
Substances - The A2 gene doesn’t convert the H3&H4,
to A very well
A much none some - The results is fewer A2 antigens sites
B none much some compared to many A1 antigen sites.
O none none much A1 and A2 SUBGROUPS

AB much much
HEMAGGLUTINATION INHIBITION
Sec+Abs+Ags on RBCs = No
agglutination(saliva)
Neutralization or inhibition
• Ex. A+Anti A= agglutination
A in secretion = Anti A+ A cells = no agglutination
B in secretion+ Anti B+ B cells + No agglutination
H in secretion+Anti H+ O cells = No agglutination

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OTHER A SUBGROUPS BOMBAY
• There are other additional subgroups of A • The hh causes NO H ANTIGEN to be
- Aint(intermediate),other A produced
• A3 red cell cause mixed field agglutination • Results in RBCS with no H,A or B
when polyclonal anti A or Anti A, B is used antigen (patient Type as O)
• Mixed field agglutination appears as small • Bombay RBCS are not agglutinated
agglutinates with a background of with Anti A, Anti B or Anti II
unagglutinated RBCs Group O RBC cannot be given because they
• They may contain Anti A still have the II antigen, so we have to
SUBGROUP transfuse the patient with blood that contains
• Use of lectins no H antigen
• Use of Anti A (group B) CLASSIFICATION OF BOMBAY (Oh)
• A1- 80% reacts with Anti A and Anti A1 1. Classical Bombay
2. H deficient Bombay Non-secretors
• A2-20% reacts with Anti A
• Characteristics
CHARACTERISTICS OF Anti A1
a. No H but A and B Ags on cells
1. Found in A2 person- 1 to 8%
b. Notations (Ah,Bh ABh on cells)
2. Found in A2B- 22 to 35%
- Charact: weak reaction with Anti A and
3. Significant only at 37 degree
Anti B
4. Causes X- matched incompatibilities
COMPARISON OF A1 AND A2 • H deficient Bombay secretors
- Inheritance f double dose of Z gene
Points of Diff A1 A2 (ZZ)
Antigen A1 A A - No H on cells but with ABH in
Present secretions
A emzyme Increased Decreased • H deficient Bombay secretors
(dominant)
Potency More potent Less - Most recent
potent - H deficient on red cells
Reactivity D.b U.e - Wk expressions of ABH secretions
(-) to U.e (+) to U.e - Alternate genes Zz
+ to Anti A plus Anti (-) to D.b. Chimerism (Blood Chimeras)- sa kambal
A1 True chimerism ( twin)
F1----F2
LECTURE 4 “A” “B” (AB no anti A= AB and Anti B)
BOMBAY PHENOTYPE “B” – contains Anti A
• Bhende 1952 in Bombay,Marathi,India Immune tolerance
• Bombay is now Mumbai Poor antigenicity
• Very rare *Blood factor was transferred to fetus two
little by little that is why there is poor
- Hh/HH (99.99) > L fucosyl transferase>
antigenicity
“H” antigen
- Hh/hh (0.01%) does not > no “H • Chimerism without a twin is called
Dispermy (Mosaicism) – egg fertilized
- Lft (OH)
by 2 sperms
- Another name- “H null” ( genotype hh)
Artificial chimeras
CHARACTERISTICS of Oh
1. Group “O’ > A or B transplant
1. No ABH
2. Exchange transfusion, group “O” > A or
2. Antibodies : Anti A, Anti B and Anti H
B
3. Enzyme: Absent
3. Feto maternal bleeding
4. I receptors: present, reacts with anti A
4. Fetal RBCs in mother’s blood
5. Anti H: igM
TYPES OF ANTIGEN THAT GIVE
6. Occurrence: consanguineous marriage
PROBLEMS IN ABO GROUPING AND
7. Inheritance: recessive mode of inheritance
CROSS MATCHING
Analysis
1. Acquired A antigen ( mistyped as A)
• Donors cell (Oh) <Anti A, < Anti B,< Anti
- O (A) and B (AB)
H,<Anti A,B (-)
Reasons: severe infection (P. mirabilis)
• Donor’s serum (oh) has anti H <A cells,<B 2. Acquired B antigen (mistyped as AB)
cells,<AB cells,<O cells all are positive
- At patients (A,B) not found in A2 and O
- Absorption of bacterial

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ALTERED ANTIGENS DEPRESSION OF ABH antigens
Types of Polyagglutination (Antigenic weakening)
• Anti T natural occurring 1. Leukemia
• Anti-Tn rarely reacts with erythrocytes 2. Lymphoma
that may have inherited or acquired 3. Refractory anemia
surface abnormalities WEAKENING OF AB ANTIGENS (A3B)
• Anti CAD throcytes that may have 1. A2B and A3B (B enzyme is more
inherited or acquired surface efficient than A enzyme)
abnormalities Ex:
Reasons why polyagglutinable cells react - A1B (A1 gee and B gene are equal in
with the antibodies efficiency by adding I, sugars to H
- Have acquired antigenic reactivity bearing chains)
- Have inherited an unusual CAD - A2B( B gene specified transferase is
“T” (antigen) activation(unmasking of more efficient H bearing chains receive
hidden antigen) more galactose)
- Temporary condition due to removal of MICTURES OF BLOOD (ABO mos
sialic acid because of the action of an phenotype)
enzyme neuraminidase • Reasons: Alleles of the ABO locus are
Organisms produce the enzyme found to be responsible
- Bacterioides Locus “O” gene  H ag Anti A and Anti B
- Protozoa “A” gene  A ag  Anti B
- Yeast • Reaction: Typed as “A” (+) with Anti A
- Viruses (+) with Anti H
REACTIONS ABH Substances in Disease
- Anti T- normal adult serum (+) 1. Pseudoantigens as in leukemi
- Anti T- cord serum (-) 2. C.I.I
- Anti R person’s own serum (-) with 3. Lymphomas
polybrene (-) 4. Immunodeficiency disease
Tn activation 5. 3-6 months and elderly
- Permanent condition due to insufficient 6. Acquired B phenomenon
sialic acid during erythrocyte maturation 7. Non septicemic P. mirabilis
Conditions associated with Tn activation
1. Hemolytic anemia
2. Leukopenia
3. Thrombocytopenia
Reactions
- Dolichos biflorus (+)
- Salvia sclarea (+)
- Salvia haemtodes (+)
- Salvia horminum (+)
- With enzymes (destroyed)
CHARACTERISTICS SHARED BY Tn
and T ACTIVATED CELLS
1. Decreased of sialic acid during
erythrocyte maturation
2. Decreased of M and N surface antigens
3. Mixed field pattern with all normal
adult sera
4. No agglutination with cord serum
5. No agglutination with polybrene
CAD POLYAGGLUTINABILITY
• Reactions: D. biflorus (+), S.horminum
(+),
• Enzymes (not destroyed)
• Enhanced by enzymes

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