ABO BLOOD TYPE INCOMPATIBILITY

Limoncito Madrid Mangaba Manlapaz Miranda Mones Montemayor Section 1 D

1. Illustrate and describe the RBC membrane.

2. Explain the role of genetics in determining the blood type, specifically in the oligosaccharide content of the red cell membrane s glycolipid.

3. Explain the oligosaccharide sequence in the RBC membrane of the different blood types. (A, B, AB, O, Bombay blood type O) BLOOD GROUP O inherit at least one genotype (HH or Hh) and two O genes. H gene elicits production of -2-L-fucosyltransferase which transfers the sugar L-fucose to an oligosaccharide chain on the terminal galactose of type 2 chains. L-fucose is the sugar responsible for H specificity or the blood group O

BLOOD GROUP A The A gene (AA or AO) codes for the production of -N-acetylgalactosaminyl transferase which transfers an N-acetyl-D-galactosamine sugar to the H substance.

BLOOD GROUP B Individuals with blood group B inherit B gene (BB or BO) that codes for the production of -3-Dgalactosyltransferase which attaches D-galactose sugar to the H substance. This is responsible for B specificity (blood group B).

BLOOD GROUP AB inherit both A and B genes. So they both have D-galactose and N-acetyl-D-galactosamine attached to H substance.

BOMBAY PHENOTYPE It represents the inheritance of a double dose of the h gene producing the very rare genotype hh. No H antigens formed; therefore, no A or B antigens formed. Phenotypes as blood group O Anti-A, anti-B, anti-A, B and anti-H present in the serum Can only be transfused with blood from another Bombay

4. Discuss the biomedical basis of blood determination. What are the different blood types and the immunodominant sugars present in blood type?

A and B antigens are oligosaccharides. The most abundant structures onred cells carrying ABO activity are the N-linked oligosaccharides of red cell surface glycoproteins, predominantly the red cell anion exchanger (AE1, the Diego blood group antigen, or band 3) and the glucose transporter (GLUT1), although some other glycoproteins are also involved. ABO-active oligosaccharides are also present on glycolipids. Oligosaccharides are chains of monosaccharide sugars: D-glucose (Glc); D-galactose (Gal); Dmannose (Man); N-acetyl-D-glucosamine (GlcNAc); N-acetyl-D-galactosamine (GalNAc); L-fucose (Fuc). An oligosaccharide is A-active when the terminal monosaccharide is GalNAc, in _1 3 linkage to a Gal residue that also has Fuc in _1 2 linkage, whereas an oligosaccharide is B-active when the terminal monosaccharide is Gal, in 1 3 linkage to the _1,2-fucosylated Gal residue (Fig. 3.2). GalNAc and Gal are the immunodominant sugars of A and B antigens, respectively. Group O red cells lack both GalNAc and Gal from the _1,2-fucosylated Gal residue (Fig. 3.2), so express neither A nor B. The A and B trisaccharidesmay be attached to several different core oligosaccharide chains, but in red cells the fucosylated Gal residue is usually in _1 4 linkage to GlcNAc (Fig. 3.2). This is called a type 2 core structure. Less abundantcore structures, called type 3 and type 4, are only present on glycolipids and may also be involved A and B activity. Type 3 and type 4 structures express A antigen on A1 phenotype red cells, but not on A2 cells, which may account for the qualitative differences between A1 and A2.

5. Discuss and compare direct and indirect blood typing; major and minor cross matching. DIRECT AND INDIRECT BLOOD TYPING Testing to detect ABO incompatibility between a donor and potential transfusion recipient is the foundation on which all other pretransfusion testing is based. ABO forward and reverse grouping tests are required to be performed on all donors and patients. Forward grouping (front type/ direct blood typing) is defined as using known sources of commercial antisera (anti-A and anti-B) to detect antigens on an individual s red blood cells.

ABO Forward Grouping: Principle- Detection of Antigens on Patient s RBCs with known Commercial Antisera Patient RBCs with Anti-A 0 4+ 0 3+ Patient RBCs with Anti-B 0 0 4+ 3+ Interpretation of Blood Group O A B AB

+ = visual agglutination 0 = negative Reaction grading vary from patient to patient On the other hand, reverse grouping (back type/ indirect blood typing) is defined as detection of ABO antibodies in the patient s serum by using known reagent red blood cells; namely A1 and B cells.

ABO Reverse Grouping: Principle- Detection of ABO Antibodies (Isoagglutinins) in Serum of Patient with known Commercial RBCs Patient Serum with Reagent A1 Cells 4+ 0 3+ 0 Patient Serum with Reagent B Cells 4+ 3+ 0 0 Interpretation of Blood Groups O A B AB

+ = visual agglutination 0 = negative Reaction grading vary from patient to patient

Summary of Forward and Reverse Groupings Forward Group Patient s Cells with Reagents Anti- A Anti-B 0 4+ 0 3+ 0 0 4+ 3+ Reverse Group Patient s Serum with Reagents A1 Cells B Cells 4+ 0 3+ 0 4+ 2+ 0 0

Blood Group O A B AB

Antigen(s) on RBCs No A or B antigen on cell A antigen on cell B antigen on cell A and B antigen on cell

Antibodies in serum A and B antibodies in serum B antibodies in serum A antibodies in serum No A and B antibodies in serum

6. Discuss the biochemical basis of ABO incompatibility.

7. Differentiate Rh Incompatibility and ABO Incompatibility. During pregnancy, red blood cells from the unborn baby can cross into the mother's bloodstream through the placenta. If the mother is Rh-negative, her immune system treats Rh-positive fetal cells as if they were a foreign substance and makes antibodies against the fetal blood cells. These anti-Rh

antibodies may cross back through the placenta into the developing baby and destroy the baby's circulating red blood cells. When red blood cells are broken down, they make bilirubin. This causes an infant to become yellow (jaundiced). The level of bilirubin in the infant's bloodstream may range from mild to dangerously high. Because it takes time for the mother to develop antibodies, firstborn infants are often not affected unless the mother had past miscarriages or abortions that sensitized her immune system. However, all children she has afterwards who are also Rh-positive may be affected. Rh incompatibility develops only when the mother is Rh-negative and the infant is Rh-positive. A, B, and O are the three major blood types. The types are based on small substances (molecules) on the surface of the blood cells. In people who have different blood types, these molecules act as immune system triggers (antigens). Each person has a combination of two of these surface molecules. Type O lacks any molecule. The different blood types are: Type A, Type B, Type AB and Type O. People who have one blood type form proteins (antibodies) that cause their immune system to react against other blood types. Being exposed to another type of blood can cause a reaction. This is important when a patient needs to receive blood (transfusion) or have an organ transplant. The blood types must be matched to avoid an ABO incompatibility reaction. For example: A patient with type A blood will react against type B or type AB blood; A patient with type B blood will react against type A or type AB blood; A patient with type O blood will react against type A, type B, or type AB blood. Because type O lacks any surface molecules, type O blood does not cause an immune response. This is why type O blood cells can be given to patients of any blood type. People with type O blood are called "universal donors." However, people with type O can only receive type O blood. Since antibodies are in the liquid part of blood (plasma), both blood and plasma transfusions must be matched to avoid an immune reaction.

References: Daniels, G., & Bromilow, I. (2007). Essential Guide to Blood Groups. Massachusetts: Blackwell Publishing Ltd. Dean ,L. (2005). Blood Groups and Red Cell Antigens. USA: National Center for Biotechnology Information