Molecular Basis for ABO Blood Types

Sept, 15, 2015Yannan Liu

Yida LiMichele Zhang

PHM142 Fall 2015 Instructor: Dr. Jeffrey Henderson

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-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

• 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-1,2-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).

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 inhemostasis/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, 453-465 (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

ReferencesD. 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, 1691-1695 (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