1. State the Landteiners principle of the ABO blood groups.

If an antigen (Ag) is present on a patients red blood cells the corresponding antibody (Ab) will not be present in the patients plasma, under normal conditions.Landsteiner, knowing that none of his subjects had been immunized, realized that natural antibodies must develop which are directed against antigens not present on the red cells. Individuals with A antigens on their red cells had sera containing Anti -B antibody. Individuals with B antigens had sera containing Anti-A. AB individuals had sera with no ABO antibodies present and O individuals sera contained Anti -A and Anti-B. Sera from group O individuals may contain a separate antibody, Anti-A,B. Anti-A,B possesses serologic activity not found in mixtures of Anti-A and AntiB. Anti-A,B sera will agglutinate A, B, and AB cells. It is particularly useful in detecting weak A and B antigens. References : http://quizlet.com/19405684/history-of-abo-import-of-karl-landsteiners-rule-flash-cards/ http://www.medialabinc.net/spg117648/the_history_of_the_abo_system_continued.aspx 2. Describe and illustrate the development of the H antigen, A antigen and B antigen. The formation of ABH antigens results from the interaction of genes at three separate loci (ABO, Hh, and Se). These genes do not actually code for the production of antigens but rather produce specific glycosyl transferases that add sugars to a basic precursor substance. A, B, and H antigens are formed from the same basic precursor material (called a paragloboside) to which sugars are attached in response to specific enzyme tranferases elicited by an inherited gene. The precursor substance on erythrocytes is referred to as type 2. This means that the terminal galactose on the precursor substance is attached to the N-acetylglucosamine in a beta 1 4 linkage .A type 1 precursor substance refers to a beta 1 3 linkage between galactose and Nacetylglucosamine. ABH antigens on the RBC are constructed on oligosaccharide chains of type 2 precursor substance. The ABH antigens develop as early as the 37th day of fetal life but do not increase much in strength during the gestational period. The RBCs of the newborn have been estimated to carry anywhere from 25 to 50 percent of the number of antigenic sites found on the adult RBC. As a result, reactions of newborn RBCs with ABO reagent antisera are frequently weaker than reactions with adult cells. The expression of A and B antigens on the RBCs is fully developed by 2 to 4 years of age and remains constant for life. As well as age, the phenotypic expression of ABH antigens may vary with race, genetic interaction, and disease states.

Reference: nd Harmening, D. (1993). Modern Blood Banking and Transfusion practices (2 Edition)

3. Where are the ABO antigens located (distribution) in the body? The ABO (H) antigens are found not only on the red cells, but also on other blood cells (endothelial cells, platelets, lymphocytes, and epithelial cells), in most body fluids (except CSF) and on the cell membranes of tissues such as intestine, urothelium and vascular endothelium. The expression of ABO(H) antigens on the red cell membrane and tissue membranes is controlled by the Hh gene. The expression of ABO(H) antigens into body fluids is controlled by Sese genes. The Sese genes, similar with Hh gene, are located ate chromosome 19q13.3; However, they are not part of the ABO system. The dominate Se gene codes for H transferase type 1 [(1,2) fucosyl-transferase; FUT2]. Without the prior addition of a fucose to the oligosaccharide chains, A and B antigens would not be expressed in the body secretions, irrespective of the presence of A and B transferases. Reference: Greer, J. Wintrobes Clinical Hematology. Volume 1. Page 636 nd Harmening, D. (1993). Modern Blood Banking and Transfusion practices (2 Edition) 4. What is the use of the anti-A, B reagent in ABO typing? Give the reactions of the normal ABO blood types, and variant groups (subgroups) in this reagent. The determination of an ABO blood group is defined by demonstrating the presence or absence of antigens A and/or B on the surface of human red blood cells and by detecting the presence or absence of anti-A and/or anti-B antibodies in the plasma. It is therefore appropriate to identify the erythrocyte antigens using known anti-A and anti-B, then to confirm the results by verifying the presence of the corresponding antibodies in the plasma from the test blood using known red blood cells A1 and B (reverse group). Additional testing of the red blood cells with Anti-A,B reagent facilitates the recognition of certain weak subgroups and is sometimes used as further confirmation of the reactions obtained with Anti-A and Anti-B reagents. Reaction of Cells Tested With Anti-A 0 + 0 + Anti-B 0 0 + + Red Cell ABO Group O A B AB Reaction of Serum Tested Against A1 Cells + 0 + 0 B Cells + + 0 0 Reverse ABO Group O A B AB

Reference: http://faculty.madisoncollege.edu/mljensen/BloodBank/lectures/abo_blood_group_system.htm http://www.fda.gov/downloads/BiologicsBloodVaccines/BloodBloodProducts/ApprovedProducts/LicensedPro ductsBLAs/BloodDonorScreening/BloodGroupingReagent/ucm081307.pdf 5. What is the anti-A reagent? Differentiate the reactions of A1 and A2, A1b and A2b with this reagent. Differences Reaction in anti-A Reaction in anti-A1 Cells Enzyme A1 + + 80 % of all group A (or AB) A1 gene elicits production of high concentrations of the enzyme _-3A2 + 20% of all group A (or AB)

N-acetylgalactosaminyltransferase, which converts almost all of the H precursor structure to A1 antigens on the RBCs. Number of antigen sites 810,000 to 1,170,000 sites on the 240,000 to 290,000 antigen adult RBC sites on the adult A2 RBC Immunodominant sugar N-acetyl-D-galactosamine N-acetyl-D-galactosamine Qualitative differences also exist, inasmuch as 1 to 8 percent of A2 individuals produce anti-A1 in their serum, and 22 to 35 percent of A2B individuals produce anti-A1. This antibody can cause discrepancies in ABO testing and incompatibilities in crossmatches with A1 or A1B cells. Because anti-A1 is a naturally occurring IgM cold antibody, it is unlikely to cause a transfusion reaction because it usually reacts better or only at temperatures well below 37_ C. It is to be considered clinically significant if it is reactive at 37_C. There must be some difference between the antigenic structure of A1 and A2 because, even though the same immunodominant sugar (N-acetyl-D-galactosamine) is attached by the same transferase (_-3-N-acetylgalactosaminyltransferase), A2 and A2B individuals cannot recognize the A1 antigen as being part of their own RBC makeup and are immunologically stimulated to produce a specific A1 antibody that does not cross-react with A2 RBCs. The antigens present on the RBCs of A1 and A2 individuals can be represented in two ways. It is generally presented that A1 has both the A and A1 antigen on the RBC, whereas A2 has only A antigen on its. However, to simplify the concept, one can think of A1 as having only A1 antigen sites and A2 as having only A antigen sites. Serum from group B individuals contains two antibodies, anti-A and anti-A1; therefore, this antibody mixture reacts with both A1 and A2 RBCs because both cells have the A antigen. If serum from a group B individual (which contains anti-A and anti-A1) is adsorbed with A2 cells (which contain only the A antigen), anti-A will bind to the RBC. The serum left after the cells and attached anti-A are removed by centrifugation is referred to as absorbed serum and contains anti -A1. This absorbed serum will react only with A1 antigen sites. The seed of the plant Dolichos biflorus, which serve as another source of anti-A1, is known as anti-A1 lectin. Lectins are seed extracts that agglutinate human cells with some degree of specificity. This reagent agglutinates A1 (or A1B) cells but does not agglutinate A2 (or A2B cells). Reference: nd Harmening, D. (1993). Modern Blood Banking and Transfusion practices (2 Edition)