Introduction To The ABO Blood Group - University Of Utah
Introduction to the ABO Blood GroupJustin R. Rhees, M.S., MLS(ASCP)CM, SBBCM
Objectives1. Describe the biochemistry and production of the A,B, and H antigens.2. Compare and contrast the subgroups of the A and Bblood types.3. Describe two lectins that can be used to aid incorrect ABO typing.4. Given the results of forward and reverse ABO typing,correctly interpret the patient’s ABO group andidentify patterns of discrepancy.
ABO TypingAnti-AAnti-BRed blood cellsHemagglutinationNo HemagglutinationAnti-A: Anti-B: 0A antigen detected
ABO TypingAnti-AAnti-BAnti-A: 0Anti-B: 0Neither A nor B antigens detected
ABO TypingAnti-A: Anti-B: A and B antigens detected
Forward Type Detection of antigens on the patient’s redcells:Anti-AAnti-BType 0A
Forward Type Detection of antigens on the patient’s redcells:Anti-AAnti-BType0 B
Forward Type Detection of antigens on the patient’s redcells:Anti-AAnti-BType AB
Forward Type Detection of antigens on the patient’s redcells:Anti-AAnti-BType00O
Reverse TypingIgGPlasma or serum(Contain antibodies)IgAIgMErythrocytes(Express antigens)
Reverse TypingAntibodies to Type BIgGPlasma or serum(Contain antibodies)IgAIgMErythrocytes(Express antigens)Type A
Reverse TypingAntibodies to Type AIgGPlasma or serum(Contain antibodies)IgAIgMErythrocytes(Express antigens)Type B
Reverse TypingPlasma or serum(Contain antibodies)Erythrocytes(Express antigens)Do not form antibodies toA or B antigensType AB
Reverse TypingAntibodies to both A and BIgGPlasma or serum(Contain antibodies)IgAIgMErythrocytes(Express antigens)Type O
Forward and Reverse TypeForward TypeAnti-AagainstPt. RBCAnti-BagainstPt. RBC
Forward and Reverse TypeForward TypeAnti-AagainstPt. RBCAnti-BagainstPt. RBCReverse TypePt. PlasmaagainstA RBCPt. PlasmaagainstB RBC
Forward and Reverse TypeForward TypeReverse TypeAnti-AagainstPt. RBCAnti-BagainstPt. RBCPt. PlasmaagainstA RBCPt. PlasmaagainstB RBCInterp. 00 A
Forward and Reverse TypeForward TypeReverse TypeAnti-AagainstPt. RBCAnti-BagainstPt. RBCPt. PlasmaagainstA RBCPt. PlasmaagainstB RBCInterp.0 0B
Forward and Reverse TypeForward TypeReverse TypeAnti-AagainstPt. RBCAnti-BagainstPt. RBCPt. PlasmaagainstA RBCPt. PlasmaagainstB RBCInterp. 00AB
Forward and Reverse TypeForward TypeReverse TypeAnti-AagainstPt. RBCAnti-BagainstPt. RBCPt. PlasmaagainstA RBCPt. PlasmaagainstB RBCInterp.00 O
ABORhAnti-AAnti-B Anti-D (Rh) A1 Cell B Cell
Basic ABO Biochemistry:ABH antigen formation Mendelian– A and B are codominant Chromosome 9– Over 200 alleles have been identified at the ABOlocus! O gene is an amorph– O/O inheritance produces O phenotype
Inheritance question My mother and father are both A positive My sister is O negative Possible?AOAA/AA/OOA/OO/O
Inheritance question My mother and father are both A positive My sister is O negative Possible?AOAA/AA/OOA/OO/ODdDD/DD/ddD/dd/dD Rh antigend lack of Rh antigen
Basic ABO Biochemistry:ABH antigen formation Glycosyltransferases: add sugars to a basicprecursor substance. 37th day of fetal life. Neonate: 25-50% antigen sites on RBC How are these antigens formed?
D-Galactose (GAL)N-acetylglucosamine (GLNAC)D-Galactose (GAL)GlucoseType 2 Precursor ChainRBC membraneCeramide
H antigenD-Galactose (GAL)N-acetylglucosamine (GLNAC)L-FucoseD-Galactose (GAL)GlucoseRBC membraneCeramide
A geneN-acetylgalacosaminyl transferaseN-acetylgalactosamineD-Galactose (GAL)N-acetylglucosamine (GLNAC)L-FucoseD-Galactose (GAL)GlucoseRBC membraneCeramide
B geneD-galactosyl transferaseD-GalactoseD-Galactose (GAL)N-acetylglucosamine (GLNAC)L-FucoseD-Galactose (GAL)GlucoseRBC membraneCeramide
O/O genes?Result: Lots of unmodified H antigens on the RBCD-Galactose (GAL)N-acetylglucosamine (GLNAC)L-FucoseD-Galactose (GAL)GlucoseRBC membraneCeramide
hh genotype The H gene is present in more than 99.99% ofthe population. (HH or Hh) The hh genotype is therefore extremely rare. Known as Oh or the “Bombay” phenotype,(hh) individuals may inherit ABO genes, butbecause the H antigen is not formed, no ABOexpression can occur.Genes: h/h, A/BNeither A nor B antigens detected
Oh Bombay phenotype First reported by Bhende in 1952 in Bombay, India.Approx 130 cases worldwide have been reported. because of hh inheritance, ABO cannot be expressed. No reactions with anti-A, anti-B, or anti-H Bombay individuals produce anti-A, anti-B, anti-A,B,and anti-H. They ABO type as O, but cannot receive Oblood. Why?– A: Type O has the highest amount of H. Transfusion oftype O blood would cause an immediate hemolytictransfusion reaction.Oh individuals should only receive Oh donor blood
Anti-H lectin A lectin is a proteinthat is capable ofbinding to acarbohydrate. A lectin with anti-Hspecificity can bederived from theseeds of the Ulexeuropaeus plantCommon gorse, Ulex europaeusPhoto credit: Creative x europaeus flowers.jpgAnti-H lectin will agglutinate Group O cells, but not Oh (Bombay) cells
Early transfusion attempts 1667 Jean-Baptiste Denis transfused bloodfrom a calf into “madman” Antoine Mauroy.Image source: Wellcome LibraryAttribute: y blood transfusion
Last half of 19th CenturyJ. H. Aveling ‘Immediate Transfusion’Image Source: Wellcome CollectionImage Source: Science Museum, LondonPhoto and Image Attributions: y blood transfusion 1873, F. Gesellius estimated that 56% oftransfusions ended in death
Karl Landsteiner Karl’s serumagglutinates mycells. My serum does notagglutinate Karl’scells. What are thepossible bloodtypes?
Karl Landsteiner Karl’s serumagglutinates mycells. My serum does notagglutinate Karl’scells. What are thepossible bloodtypes?Karl is type O. I am type A, B, or ABKarl is type A or B. I am type AB
A Subgroups A subgroups:– A1 A2 A3 Ax Aend Am Ay Ael etc. Approx. 80% of type A individuals are A1 Approx. 20% of type A individuals are A2 The remaining subgroups comprise 1%
A1 and A2 Inheritance of an A1 gene elicits production ofhigh concentrations ofα-3-N-acetylgalactosaminyltransferase Converts almost all of the H precursorstructure to A1 antigens. A1 antigens are more highly branched than the“common A” structure shown previously A2 type has fewer antigens per cell, only existas “common A”
A1 and A2 The immunodominant sugar on both A1 andA2 RBCs is N-acetyl-galactosamine; however,there are subtle antigenic differences whichcause the body to discern self from non-self. A1: 810,000 to 1,170,000 antigen sites A2: 240,000 to 290,000 antigen sites
A1 and A2 A1 subgroup has both“common A” and A1antigens. Most of the Hantigens have beenconverted. A2 subgroup has only“common A” antigens.More unconverted Hantigens.A A1A1A1 typeA1 AAAAA1AA1A2 typeAA “Common A” antigensA1 highly branched A antigens
Anti-A1 Because approximately 20% of type Aindividuals are A2, we sometimes encounteranti-A1 in transfusion medicine. Anti-A1 is non-RBC Immune, IgM, and usuallycold reacting. It is only considered clinicallysignificant if it is reactive at 37 C. Anti-A1 is produced by approx. 1-8% of A2individuals.
ABO DiscrepancyForward TypeReverse TypeAnti-AAnti-BA1 CellB Cell 0
Anti-A1 Lectin A purified extract made from the seeds of the Dolichos biflorusplant agglutinate red blood cells with A1 antigens present. Note: there is no anti-A2 lectin. Why?BloodGroupA1A2AntigenPresentA1AAAnti-AAnti-A1 Lectin(Anti-A plusAnti-A1) 0
Weak subgroups of A As stated before, the prevalence of A subgroupsof A weaker than A1 and A2 is less than 1%SubgroupLaboratory ResultsNumber of Aantigenic sitesA3Mixed field reaction with anti-A and most anti-A,Breagents35,000 perRBCAxCharacteristically not agglutinated with anti-A butdo agglutinate with most examples of anti-A,B4000AendMixed field reaction with anti-A and anti-A,B. Aendis inherited as an allele at the ABO locus. Anti-A1is found in some sera. Only H is found insecretions.3500AmCharacteristically not agglutinated, or very weaklyagglutinated by all anti-A and anti-A,B reagents.Usually do NOT produce anti-A1 in sera.200-1900
Weak subgroups of A Fewer antigen sites on the RBC means weakerreactions with antisera. It is possible for an Ax donor to be mistyped asO. This unit could then be transfused into anO recipient, who has anti-A,B. The anti-A,Bantibody in the recipient could agglutinateand lyse the donor Ax RBCs and causeintravascular hemolysis.
Weak subgroups of B Subgroups of B are very rare and less frequentthan A subgroups.– B, B3, Bx, Bm, Bel, etc.
AB subgroups AB individuals can demonstrate subgroups ofA, B or both– A1B, A2B, AxB, A1Bel, etc.
Reactivity of anti-H lectinO A2 B A2B A1 A1B Oh (Bombay)Greatest amount of HLeast amount of H
ABO Discrepancies All ABO Discrepancies must be resolvedprior to reporting a patient or donor ABOgroup. Why investigate these discrepancies?
Forward TypeIs there anything wrong with thispicture?
Mixed Field (MF) ReactionControl tubesPatient tubes
Case StudyA technologist reads and reports a patient’sblood type:Anti-AAnti-BAnti-DA1 CellB Cell3 3 4 00Interpretation: AB Positive
Case StudyA technologist reads and reports a patient’sblood type:Anti-AAnti-BAnti-DA1 CellB Cell3 3 4 00Interpretation: AB PositiveThe sample is from an A positive patientundergoing a type B negative BMT.Lack of visible reverse type is due toimmunosuppression.According to our protocols, the patientshould be supported on irradiated, washed,O negative red cells.
Question 1Forward TypeReverse TypeAnti-AAnti-BA1 CellB Cell00
Question 2Forward TypeReverse TypeAnti-AAnti-BA1 CellB Cell 00
Question 3Forward TypeReverse TypeAnti-AAnti-BA1 CellB Cell 00
Question 4Forward TypeReverse TypeAnti-AAnti-BA1 CellB Cell0 00
Question 5Forward TypeReverse TypeAnti-AAnti-BA1 CellB Cell 0
Question 6What do vampires put on their steak?Answer: A1Image credit: Creative CommonsAttribute: pire.svg
References Harmening DM, Ed. Modern Blood Bankingand Transfusion Practices, 6th Ed. F. A. DavisCompany, Philadelphia. 2012. Fung MK, Eder AF, Spitalnik SL, Westhoff CM.AABB Technical Manual, 19th Ed. AABB Press.2017
Converts almost all of the H precursor structure to A 1 antigens. A 1 antigens are more highly branched than the “common A”
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4 THE ABO BLOOD GROUP SYSTEM 25 4.1 The ABO Blood Groups 26 4.2 Basic Genetics of the ABO Blood Groups 27 4.3 Demonstrating ABO Blood Groups 29 4.4 Development of the A and B Red Cell Antigens and Antibodies 30 4.5 The Subgroups of the Antigen A 30 4.6 Anti-A 1 and Anti-AB in Blood Grouping Tests 31 4.7 IgM and IgG (Naturally-occurring and Immune)
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