ABO INCOMPATIBILITY

Group 6 Reporters: ESTRADA, James Raphael ESTEBAN, Laurence ESPERANZA, Wynwright Members: ESPINOSA, Kesshia ESQUIDA, Patria ESTEBAN, Emel Rose ESTRELLA, Lee Anne

LEARNING OBJECTIVES:
To describe the composition of RBC membrane To explain the importance of the oligosaccharide sequence in the RBC membrane in blood type determination To determine the role of genetics in determining the blood type, especially in the oligosaccharide content of membrane lipoproteins To illustrate the biochemical basis of blood typing and blood determination To identify the biochemical basis of ABO incompatibility

COMPOSITION OF RBC MEMBRANE

The Red Cell Membrane

transport of respiratory gases to and from the tissues

Primary function

Extremely tough yet highly flexible

Functions
Maintains the characteristic shape of the red cell Acts as the interface between the cell and its environment via membrane surface receptors

Separates the contents of the cell from the plasma

Regulates intracellular cation concentration

The red cell membrane consists of:

Composition
Membrane Carbohydrates Occur only on the external surface of the red cell as: Glycoprotein Glycolipids Ex. ABO blood group (antigen)

Composition

Composition
MEMBRANE PHOSPHOLIPIDS Polar head group hydrophilic(water loving) Non-polar fatty acid tail (Hydrocarbon chains) hydrophobic (water fearing)

Phospholipids

Choline phospholipids (extracellular layer)

Amino phospholipids (cytoplasmic layer)

Phosphatidyl choline

Sphingomyelin

Phosphatidyl ethanolamine

Phosphatidyl

serine

Phosphatidyl Ethanolamine

Phosphatidyl Serine

MEMBRANE CHOLESTEROL
Unesterified Lies between the 2 layers of the lipid bilayer

MEMBRANE CHOLESTEROL
MEMBRANE CHOLESTEROL

surface area

deformability

premature RBC destruction (in extreme circumstances)

Membrane Proteins
Peripheral Integral Named according to their relative positions on SDSPAGE electrophoresis

Membrane Proteins

PERIPHERAL PROTEINS
Interact to form a cytoskeleton. acts as a tough supporting framework for the lipid bilayer.

FOUR peripheral proteins play a key role in the structure of the red cell cytoskeleton:
Spectrin Ankyrin Protein 4.1 Actin

peripheral proteins
Spectrin (Bands 1 and 2) Most abundant consists of : - chain - chain Responsible for the flexibility of the RBC membrane Ankyrin(Bands 2.1-2.3) This serves to anchor assembled spectrin molecules to the lipid bilayer.

peripheral protei ns
Actin(Band 5) Globular protein Composed of filaments The filaments bind weakly to the tail end of both and spectrins. Band 4.1 Globular protein Binds to spectrin close to the actin binding site thereby strengthening and stabilizing the cytoskeletal lattice. Also binds directly to Glycophorins A and C and Band3. Strengthens the links between the lipid bilayer and the protein cytoskeleton.

Integral Proteins
These penetrate the lipid bilayer and are firmly anchored within it. -Band 3 -Glycophorins A, B, and C. -Na+/K+ ATPase. -glucose transport protein. -surface receptors.

Anion Exchange Protein(Band 3)

Single molecule Molecular wt.- 95000 25% of total protein content 2 major functions: Facilitates anion transport via the red cell membrane Important binding site for cytoskeletal and other red cell proteins

Glycophorins
3 members of the RBC glycophorin family: Glycophorins-A,B and C Sialoglycoproteins (combination of a sugar and protein) Act as :
transmembrane signal transducers receptor for the Plasmodium falciparum protein PfEBP 2(erythrocyte binding protein 2)

Na+/K+ATPase
Catalyses the hydrolysis of ATP to ADP Each ATP molecule hydrolysed: Ejection of 3 Na+ ions transport of 2 K+ ions into the cell

Glucose transport Protein

Molecular wt.- of 60 000 ATP hydrolysis - not required Motive force for transport of plasma Glucose into the red cell - derived from the electrochemical gradient of Na+ ions across the cell membrane accompanied by a Na+ ion Leading to a net reduction in the transmembrane gradient of Na+ ions. Failure of the cation pump to regenerate the Na+ gradient ,would result in failure of Glucose transport. May lead to glycolytic failure and hence lack of ATP generation. Final result-cell death.

Surface receptors
Most important surface receptor is transferrin receptor. Highest concentration on RBC surface. The receptor domain is capable of binding two transferrin molecules. Receptor-transferrin complexes are internalised. Iron released from the transferrin

IMPORTANCE OF THE OLIGOSACCHARIDE SEQUENCE IN THE RBC MEMBRANE IN BLOOD TYPE DETERMINATION

OLIGOSACCHARIDE

3-12 monosaccharide Attach to the proteins and lipids on the surface of erythrocytes

Immunodominant sugar: N-acetylgalactosamine

Immunodominant sugar: Galactose

ROLE OF GENETICS IN DETERMINING THE BLOOD TYPE

Type 2 Precursor Chain

Paragloboside Common carbohydrate residue Genetic product Type 2 refers to the (1-4) glycosidic linkage between terminal Gal and GlcNAc Type 1 refers to (1-3) glycosidic linkage restricted to epithelia, in the gastrointestinal or reproductive tracts.

H Substance
Derivative of Type 2 Precursor Chain Can be inherited from a parent having either an HH or Hh genotype Elicits the production of fucosyl transferase

50% HH

HH

Hh 50% Hh

25% HH 50% Hh

Hh

Hh
25% hh
Bombay Phenotype (rare case)

ABO Substances
1. A Substance AA or AO is responsible for the production of N-acetylgalactosaminyl transferase, which attaches GalNAc to the terminal galactose of the H Substance Immunodominant sugar: N-acetylgalactosamine

AA AA

100% AA

AA Ao

50% AA

50% Ao

ABO Substances
2. B Substance BB or BO is responsible for the production of Galactosyl transferase, which attaches Gal to the terminal galactose of the H Substance Immunodominant sugar: Galactose

BB BB

100% BB

BB Bo

50% BB

50% Bo

AA BB

100% AB

AB AB

25% AA 25% BB 50% AB

ABO Substance
3. O Substance Homozygous genotype OO does not express any enzyme that would attach any additional sugar to the H Substance O Substance: No immunodominant sugar

oo

oo

100% oo

AA oo

100% Ao

AO oo

50% AO

50% OO

AB oo

50% AO

50% BO

4 4

BIOCHEMICAL BASIS OF BLOOD TYPING AND BLOOD DETERMINATION

ABO System

Karl Landsteiner 1900

Definition of Terms
Agglutinin Specific antibody in blood plasma capable of causing the clumping of RBC or bacteria or particles such as viruses Antibody which aggregates a particulate antigen Agglutinogen Antigen located on the surface plasma membrane of RBC which determines the blood group of the individual Stimulates the production of agglutinin Agglutination The clumping together of blood cells or microorganisms, usually due to an antigen-antibody reaction

ABO Blood Typing

Blood Type Antigens (Agglutinogens) on Red Blood Cells Antibodies (Agglutinins) in Plasma

A B AB O

A B A&B Neither

Anti-B Anti-A None Anti-A & Anti-B

ABO Blood Groups

Plasma

Plasma Plasma

Plasma

No Agglutination Reaction

Rh System Rh+ (+ D antigen) Dominant (DD) Heterozygous Rh (- D antigen) Homozygous recessive (dd)

(Dd)

BIOCHEMICAL BASIS OF ABO INCOMPATIBILITY

Transfusion Reactions
ABO incompatibility is the most potent cause of immediate hemolytic reactions

Hemolytic Disease of the Newborn

caused by immunization of an Rh-negative mother by the red blood cells from an Rh-positive fetus