PHM142 Fall 2015
Instructor: Dr. Jeffrey Henderson
Molecular Basis for ABO Blood
Types
Sept, 15, 2015
Yannan Liu
Yida Li
Michele Zhang
Historical Context
• ABO blood group system first discovered by
Karl Landsteiner in 1900
• Received Nobel Prize in Physiology or
Medicine in 1930
• Czech serologist Jan Janský first to classify
blood into 4 types (A, B, AB, 0)
• Did not receive a Nobel Prize
Landsteiner, K. (1900). Zur Kenntnis der antifermativen, lytischen und agglutinierenden Wirkungendes Blutserums und der
Lymphe. Zentbl. Bakt. Parasitkde (Abt.) 27, 357-363.
Antigen
• Antigens trigger antibody reactions
• Blood cells make isoantigens
• Your blood type is determined by the antigen
expressed by your blood cells
• Your body usually makes antibodies against
foreign bodies
US National Library of Medicine. (n.d.). Antigen. Retrieved from MedlinePlus:
https://www.nlm.nih.gov/medlineplus/ency/article/002224.htm
Antibodies
• Antibodies are made by the B cells in your
immune system
• Y-shaped proteins with 2 heavy
and 2 light chains; highly specific
• 5 types of Igs but blood types are
IgM or IgG (for O blood type)
• Isoantibodies or Alloantibodies
The McGraw-Hill Companies, inc. (n.d.). Humoral Immunity. Retrieved from
http://www.mhhe.com/biosci/esp/2001_saladin/folder_structure/tr/m4/s8/
AZoNetwork. (n.d.). What is an Antibody? Retrieved from News Medical: http://www.news-medical.net/health/What-isan-Antibody.aspx
Australian Red Cross Blood Service. (n.d.). ABO and RhD. Retrieved from Australian Red Cross Blood Service:
http://www.transfusion.com.au/blood_basics/blood_groups/abo_rh
• ABO systems comprises of 4 blood types:
• A, B, AB and O
• Blood type is determined by the antigen(s)
present on the surface of the blood cells
• These antigens are oligosaccharide variants
with a common backbone situated on
membrane glycoprotein and glycolipids
• Genes determining blood type encodes for
different glycosyltransferases responsible for
antigen differentiation
F.-i. Yamamoto, H. Clausen, T. White, J. Marken, S.-i. Hakomori, Molecular genetic basis of the histo-blood group ABO system.
Nature 345, 229-233 (1990)
Lodish, H. F.; Berk, A.; Kaiser, C. A.; Krieger, M.; Bretscher, A.; Ploegh, H.; Amon, A.; Scott, M. P.Molecular Cell Biology; 2013;
Vol. 7, p. 462.
ABO Antigen Biosynthetic Pathway
F. Yamamoto, H. Clausen, T. White, J. Marken, S.-i. Hakomori, Molecular genetic basis of the histo-blood group ABO system. Nature
345, 229-233 (1990)
• Antigen H is the product of α-1,2fucosyltransferase
• Antigen A results from GalNac attachment by
α-1,3- n-acetylgalactosaminyltransferase
• Antigen B results from Gal attachment by α1,3-galactosyltransferase
• Individuals with blood type AB expresses both
enzymes
• Individuals with type O blood contains neither
enzymes  H antigen remains unmodified
G. A. Bohmig, A. M. Farkas, F. Eskandary, T. Wekerle, Strategies to overcome the ABO barrier in kidney transplantation. Nat Rev Nephrol,
(2015).
N-acetylgalactosaminyltransferase (GTA)
J. A. Letts et al., Differential recognition of the type I and II H antigen acceptors by the human ABO(H) blood group A and B
glycosyltransferases. J Biol Chem 281, 3625-3632 (2006).
Clinical Applications
The ECO Project
• Goldstein – conversion of type B RBCs
• Exoglycosidase that could selectively remove
α-Gal of B antigens
• Isolated α-galactosidase from coffee beans
and tested these on B antigens
• The resulting B-ECO RBCs functioned normally
in Phase I clinical trials
J. Goldstein, G. Siviglia, R. Hurst, L. Lenny, L. Reich, Group B erythrocytes enzymatically converted to group O survive normally in A, B,
and O individuals. Science 215, 168-170 (1982).
The ECO Project
• Olsson et al – conversion of type A RBCs
– Type A RBCs have three subtypes
– Isolated α-N-acetylgalactosaminidase from
Clostridium perfringens
– Found a reduction in antigen expression but not
complete homogeneity
M. L. Olsson et al., Universal red blood cells—enzymatic conversion of blood group A and B antigens. Transfusion Clinique et
Biologique 11, 33-39 (2004).
The ECO Project
• Kwan et al –
Discovery of an
enzyme compatible
of cleaving both A
and B antigens
D. H. Kwan et al., Toward Efficient Enzymes for the Generation of
Universal Blood through Structure-Guided Directed Evolution.
Journal of the American Chemical Society 137, 5695-5705
(2015).
The ECO Project
– Through directed evolution, generated variants of
a glycoside hydrolase that are able to efficiently
cleave the terminal trisaccharide of A and B
antigens were isolated
– The resulting antigens do not induce a negative
response from transfusion
D. H. Kwan et al., Toward Efficient Enzymes for the Generation of Universal Blood through Structure-Guided Directed Evolution.
Journal of the American Chemical Society 137, 5695-5705 (2015).
von Willebrand Disease
• Inherited, genetic disorder characterized by a defect or
deficiency of von Willebrand Factor
• Von Willebrand factor:
• Adhesion
glycoprotein
• Involved in
hemostasis/blood
clotting
• Protects blood factor
VIII from proteolytic
degradation
J. Gill, J. Endres-Brooks, P. Bauer, W. J. Marks, R. Montgomery, The
effect of ABO blood group on the diagnosis of von Willebrand
disease. Blood 69, 1691-1695 (1987).
(Medlibes Online Medical Library), (2015).
von Willebrand Disease
• 66% of total variation in VWF levels in the
blood plasma are genetically determined
• 30% of this genetic component is based on
ABO blood groups
• People with blood type O have lower plasma
levels than other blood types
– Of a study of 114 with VWD, blood type O was
found in 77%
M. Franchini, F. Capra, G. Targher, M. Montagnana, G. Lippi, Relationship between ABO blood group and von Willebrand factor
levels: from biology to clinical implications. Thrombosis Journal 5, 14 (2007).
von Willebrand Disease
• Drugs and treatment:
– Desmopressin: vasopressin analogue that
stimulates release of VWF from endothelial
storage, and stabilizes factor VIII in blood plasma
– Replacement therapy: injection of plasma
concentrates containing VWF
– Contraceptives: use of progestin contraceptives to
reduce menstrual bleeding
– Antifibrinolytic medicines: clot stabilizing agent
M. A. Laffan et al., The diagnosis and management of von Willebrand disease: a United Kingdom Haemophilia Centre Doctors
Organization guideline approved by the British Committee for Standards in Haematology. British Journal of Haematology 167, 453465 (2014).
Summary
•
•
•
•
•
•
An antigen is any particle (foreign or not foreign) that can trigger an antibody response in the body
Antibodies are made by B cells and take part in immune responses; they are Y-shaped, have 2 heavy and 2
light chains and are highly specific
Blood types are determined by the blood antigen you possess (A type means RBC has A-antigen), and your
body rejects blood antigens you don’t possess (antibody against antigens not present in the body)
– A has A-antigen; reactive (antibodies) against B-type blood and AB-type blood
– B has B-antigen; reactive against A-type blood and AB-type blood
– AB has both A-antigen AND B-antigen; reactive against nothing (universal acceptor)
– O has no antigens; reactive against all blood types EXCEPT O (universal donor)
Blood antigen differentiation is determined by the activity of glycosyltransferases in the biosynthetic
pathway
– Antigen H (O type blood) has no additional attachments,
O negative is considered the Universal Donor
– Antigen A is due to a GalNac attachment
– Antigen B is due to a Galactose attachment
– AB type blood has both antigen A and antigen B
AB positive is considered the Universal Acceptor
Clinical applications:
– The conversion of A and B antigens to H or unreactive antigens using cleavage enzymes can possibly
eliminate blood type as a risk factor in blood transfusions, organ and tissue transplants
Applications in disease: von willebrand’s disease
– Von willebrand’s disease is characterized by defect or deficiency of von Willebrand Factor, which
plays a crucial role in blood clotting
– People with blood type O have lower levels of VWF than other blood types
– Main treatment: desmopressin
Summary
References
D. H. Kwan et al., Toward Efficient Enzymes for the Generation of Universal Blood through Structure-Guided Directed Evolution. Journal of the American
Chemical Society 137, 5695-5705 (2015).
F. Yamamoto, H. Clausen, T. White, J. Marken, S.-i. Hakomori, Molecular genetic basis of the histo-blood group ABO system. Nature 345, 229-233 (1990)
G. A. Bohmig, A. M. Farkas, F. Eskandary, T. Wekerle, Strategies to overcome the ABO barrier in kidney transplantation. Nat Rev Nephrol, (2015).
J. Gill, J. Endres-Brooks, P. Bauer, W. J. Marks, R. Montgomery, The effect of ABO blood group on the diagnosis of von Willebrand disease. Blood 69, 16911695 (1987).
J. Goldstein, G. Siviglia, R. Hurst, L. Lenny, L. Reich, Group B erythrocytes enzymatically converted to group O survive normally in A, B, and O individuals.
Science 215, 168-170 (1982).
J. A. Letts et al., Differential recognition of the type I and II H antigen acceptors by the human ABO(H) blood group A and B glycosyltransferases. J Biol Chem
281, 3625-3632 (2006).
Lodish, H. F.; Berk, A.; Kaiser, C. A.; Krieger, M.; Bretscher, A.; Ploegh, H.; Amon, A.; Scott, M. P.Molecular Cell Biology; 2013; Vol. 7, p. 462.
M. Franchini, F. Capra, G. Targher, M. Montagnana, G. Lippi, Relationship between ABO blood group and von Willebrand factor levels: from biology to
clinical implications. Thrombosis Journal 5, 14 (2007).
M. A. Laffan et al., The diagnosis and management of von Willebrand disease: a United Kingdom Haemophilia Centre Doctors Organization guideline
approved by the British Committee for Standards in Haematology. British Journal of Haematology 167, 453-465 (2014).
M. L. Olsson et al., Universal red blood cells—enzymatic conversion of blood group A and B antigens. Transfusion Clinique et Biologique 11, 33-39 (2004).
Landsteiner, K. (1900). Zur Kenntnis der antifermativen, lytischen und agglutinierenden Wirkungendes Blutserums und der Lymphe. Zentbl. Bakt. Parasitkde
(Abt.) 27, 357-363.
US National Library of Medicine. (n.d.). Antigen. Retrieved from MedlinePlus: https://www.nlm.nih.gov/medlineplus/ency/article/002224.htm
The McGraw-Hill Companies, inc. (n.d.). Humoral Immunity. Retrieved from http://www.mhhe.com/biosci/esp/2001_saladin/folder_structure/tr/m4/s8/
AZoNetwork. (n.d.). What is an Antibody? Retrieved from News Medical: http://www.news-medical.net/health/What-is-an-Antibody.aspx
Australian Red Cross Blood Service. (n.d.). ABO and RhD. Retrieved from Australian Red Cross Blood Service:
http://www.transfusion.com.au/blood_basics/blood_groups/abo_rh