1. Molecular Basis for ABO Blood Types
PHM Fall 2015
Instructor: Dr. Jeffrey Henderson
Molecular Basis for ABO Blood Types
Sept, 15, 2015
Yannan Liu
Yida Li
Michele Zhang

2. 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,

3. 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:

4. 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

5. AZoNetwork. (n. d. ). What is an Antibody
AZoNetwork. (n.d.). What is an Antibody? Retrieved from News Medical:

6. Australian Red Cross Blood Service. (n. d. ). ABO and RhD
Australian Red Cross Blood Service. (n.d.). ABO and RhD. Retrieved from Australian Red Cross Blood Service:

7. 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, (1990)

8. Lodish, H. F. ; Berk, A. ; Kaiser, C. A. ; Krieger, M. ; Bretscher, A
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.

9. 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, (1990)

10. Antigen H is the product of α-1,2-fucosyltransferase
Antigen A results from GalNac attachment by α-1,3- n-acetylgalactosaminyltransferase
Antigen B results from Gal attachment by α-1,3-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).

11. 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, (2006).

12. Clinical Applications

13. 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, (1982).

14. 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, (2004).

15. 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, (2015).

16. 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, (2015).

17. 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, (1987).
(Medlibes Online Medical Library), (2015).

18. 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).

19. 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, (2014).

20. 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

21. Summary

22. 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, (2015). F. Yamamoto, H. Clausen, T. White, J. Marken, S.-i. Hakomori, Molecular genetic basis of the histo-blood group ABO system. Nature 345, (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, (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, (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, (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 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, (2014). M. L. Olsson et al., Universal red blood cells—enzymatic conversion of blood group A and B antigens. Transfusion Clinique et Biologique 11, (2004). Landsteiner, K. (1900). Zur Kenntnis der antifermativen, lytischen und agglutinierenden Wirkungendes Blutserums und der Lymphe. Zentbl. Bakt. Parasitkde (Abt.) 27, US National Library of Medicine. (n.d.). Antigen. Retrieved from MedlinePlus: The McGraw-Hill Companies, inc. (n.d.). Humoral Immunity. Retrieved from AZoNetwork. (n.d.). What is an Antibody? Retrieved from News Medical: Australian Red Cross Blood Service. (n.d.). ABO and RhD. Retrieved from Australian Red Cross Blood Service: