Immunohematology - 05 Other Blood Group Systems

IMMUNOHEMATOLOGY GENETIC CONCEPT OF LEWIS OTHER MAJOR AND MINOR BLOOD GROUP SYSTEMS AND BLOOD GROUP COLLECTIONS • • OUTLINE • Other Major & Minor Blood Group Systems o Lewis System o MNSs U Blood Group System o P Blood Group System o I Blood Group System o Kell Blood Group System o Duffy Blood Group System o Kidd Blood Group System o Lutheran Blood Group System o Minor Blood Group o Summary of Major and Minor Blood Group System • • • The Lewis gene (FUT3) is linked to Se (FUT2) and H (FUT1), all located on chromosome 19. The synthesis of Lewis antigens depends on the interaction of the transferases produced by the Lewis and secretor genes. The Lewis and secretor transferases preferentially fucosylate type 1 chains, whereas the H gene (FUT1) preferentially fucosylates type 2 chains. Type 1 chain refers to the beta linkage of the number 1 carbon of galactose to the number 3 carbon of Nacetylglucosamine (GlcNAc) residue of the precursor structure OTHER MAJOR & MINOR BLOOD GROUP SYSTEMS LEWIS SYSTEM (Le:007) • • • • • • • • • • • • • • • • • • • • IgM Reacts at RT/immediate phase (antibody screening & crossmatching) Not clinically significant Enhanced/destroyed by enzyme Located at Short arm of chromosome 19 (Secretor locus) o H, Se genes are also found in 19 90% is (+) for Le gene RBCs: glycolipids Secretions: glycoprotein Le gene encodes L-fructosyltransferase type 3 (FuT 3) o FuT 1 - H gene o FuT 2 - Se gene Produced in tissues → go to plasma → adsorbed to RBCs Se gene: formation of Le (a+b+) or partial secretory phenotype o Seen in Polynesians, Australians and Asians Lewis (+) → Lewis (-): Abnormal lipid metabolism such as pregnancy, cancer, alcoholic cirrhosis, and viral infection Leb has receptor for Helicobacter pylori Lex is marker for Reed Sternberg cells of Hodgkin’s Disease Le, H, Se are related Two alleles: Le and le are codominant o Le - codes for transferase enzyme (FuT 3) o le gene - is an amorph (non detectable) Lewis antigens are not intrinsic to red cell membraine They are produced in secretions and adsorbed to plasma onto the red cell membraine Poorly developed at birth • • • • Le genes, similarly with ABO system, codes for the production of an enzyme Lea o FuT 3 adds fucose to the subterminal of Nacetylglucosamine residues on the precursor substance to form Lea antigen o Does not depend on the secretor gene o Only require Le gene to be produced type 1 chain Leb o FuT 3 adds subterminal of N-acetylglucosamine residues on the precursor substance t o It will depend if there is a secretor gene o FuT 2 adds fucose to the same precursor substance in secretions resulting to formation of Leb ag o Requires Le & Se genes to be produced The expression is affected by the following key-player genes: Le, Se, and H o Lewis gene - either Le or an amorph le ▪ If you inherit Le - Lea can be expressed (no need for secretor gene) ▪ If you inherit le - Lea (-) o • • Secretor gene - either Se or se ▪ If you inherit Se, and Lea (+) → Leb ▪ If you inherit Se, but no Lea → no conversion to Leb so it is Le(a-b-) or Lewis null ▪ If you inherit se, but Lea (+) → no conversion so it is Le(a+b-) or Lea ▪ If you inherit se and you don’t have Lea → nothing will happen o H gene ▪ H gene will serve as acceptor for modification A, B, H antigen ▪ If you inherit H gene, you will have ABH antigens on RBCs and secretions (if no Se gene, you will not have any on secretions ▪ If you inherit h, you will not have ABH antigens on both RBCs and secretions Development of Lewis antigens o Depending on the genes inherited, Lea and Leb glycoproteins will be present in the saliva of ewborns, but Lewis glycolipids are not detectable in the plasma until about 10 days after birth. o As a result, cord blood and RBCs from newborn infants phenotype as Le(a–b–). o Some can be shown to be weakly Le(a+) when tested with a potent anti-Lea or with methods more sensitive than direct agglutination. o Lewis antigens will start to appear shortly after birth, with Lea developing first when the Le gene is present. o The Lewis fucosyltransferase (FuT 3) is more active than the secretor fucosyltransferase (FuT 2) in newborns, so more type 1 chains are available for conversion to Lea. o As the secretor transferase activity increases, converting type 1 to type 1H, Leb will be detected. o In children who inherit both Le and Se genes, the transformation can be followed from the Le(a–b–) phenotype at birth to Le(a+b–) after 10 days to Le(a+b+) and finally to Le(a–b+), the true Lewis phenotype, after about 6 years. o In contrast, children who inherit Le and sese genes phenotype as Le(a–b–) at birth and transform to Le(a+b–) after 10 days; the Le(a+b–) phenotype persists throughout life. Individuals with lele genes phenotype as Le(a–b–) at birth and for the rest of their lives. Changes in Lewis Phenotype o Many pregnant women lose their Lewis antigens during pregnancy ▪ Common to see Lewis abs in obstetrical patients ▪ After delivery, the antibody fades and the previous Lewis phenotype return o Also seen in caner patients, alcoholic cirrhosis, viral and parasitic infections LEWIS ANTIBODIES • • • Anti Lea is the most common Anti Leb Two forms: o Anti-LebH – reacts with Leb and H antigen o Anti-LebL – reacts with Leb Anti Lex – agglutinates all le (+) individuals, formed by Le(a-b-) individuals Genotype Le(a+b-) Le(a-b+) Le(a-b-) (Lewis null) Possible Antibodies Produced Anti-Leb None (does not produce anti-Lea due to they still produce Lea in saliva) Anti-Lea, Anti-Leb LEWIS ANTIGENS • Lea - independent on Se gene, will always be produced with our without the Se gene Leb - dependent on secretor gene • • Genes Inherited Secretions Red Cell Antigens Le, Se (ABH) Lea, Leb, A, B, H Le(a-b+), A, B, H le, Se (ABH) A, B, H Le(a-b-), A, B, H * Le, sese (ABH) Lea Le(a+b-), A, B, H lele, sese (ABH) None Le(a-b-), A, B, H Le, Se (ABhh) Lea, Leb Le(a-b+) Le, sese (ABhh) Lea Le(a+b-), Oh • Notes: o H (L-fucose) acts as a link to show A (Nacetylgalactosamine) and B (D-galactose) in the RBC surface. Because A and B basic precursor is H) o * may be due to modified phenotype caused by pregnancy or cancer Substance Secretor Genes Red Cell Present in Phenotype Status Inherited Antigens Secretions AB Secretor Le, Se, H, Lea, Leb, Le(a-b+), Leb A/B/O A, B, H A, B, H AB Secretor lele, Se, A, B, H Le(a-b-), Lewis null H, A/B/O A, B, H AB NonLe, sese, Lea Le(a+b-), Lea Secretor H, A/B/O A, B, H AB Nonlele, sese, None Le(a-b-), Le null secretor H, A/B/O A, B, H BOMBAY Le, Se, Lea, Leb Le(a-b+), Leb (Oh) hh, A/B/O Oh Parabombay H, A/B/O Lea Lea(a+b-), Lea (Ah, Bh, ABh) A, B MNSs U BLOOD GROUP SYSTEM (002) MN BLOOD GROUP • • • • • IgM Reacts at RT and immediate stage Not clinically significant Destroyed by enzymes Exhibits dosage o Homozygous - strong rxn (4+), double dose (MM/NN) o Heterozygous - 2+ agglutination (MN) MNSs U ANTIGENS • • • • MNS are inherited as Located on MNS antigens are important markers in paternity testing using Found on MN ANTIGENS • • • • • • Found on a well-characterized glycophorin A (GPA) Glycophorin A - major sialic acid rich glycoprotein present on outermost portion of RBC membrane, first to be exposed and destroyed/degraded by enzymes Destroyed by routine blood bank enzyme (papain, etc.) and ZZAP + DTT + papain/ficin The amino acid residue in position ?? and ?? determine the expression of MN glycophorin A M antigen = N antigen = Ss ANTIGENS • • • • • • • Locates on smaller glycoprotein, GPB S has methionine while s has threonine easily degraded by enzymes; not destroyed by Trypsin Associated with Glycophorin B Glycophorin B- Sialic acid rich glycoprotein present on the cell membrane (a.k.a Ss Sialogycoprotein) o Found on the innermost portion of RBC surface o Not easily destroyed The amino acid residue in position 29 of glycophorin B determines the expression of Ss antigens S antigen = s antigen • • Anti-S and Anti-s o IgG (react at 37C during AHG/thermophase) o Most are warm reacting at 37C and antiglobulin phase o May bind complement and have been associated with o Incubate at RT and perform antiglobulin test w/o incubating at 37C Anti-U o Formed by S-s individuals o Common among blacks (African blacks) P BLOOD GROUP SYSTEM (P:003) & GLOBOSIDE (028) BLOOD GROUP SYSTEMS • P group antigens are synthesized by sequential action of glycosyltransferases • Found on RBCs, lymphocytes, granulocytes, and monocytes (P1, P, and Pk) • Platelet, epithelial cells and fibroblasts (P) • Echinococcus granulosus - hydatid cyst fluid o Smallest cestode Phenotypes Detectable Antigens Possible Antibodies P1 P1, P, Pk None P2 P, Pk Anti-P1 p None Anti-PP1Pk k P1k P1, P Anti-P P2k Pk Anti-P, anti-P1 P ANTIGENS U ANTIGENS • • • Also carried by Stands for universal RBCs with the S or s antigen also have the U antigen INCIDENCE OF PHENOTYPES (Whites, Blacks) • • • • MN - 50%, 44% M - 28%, 26% N - 22%, 30% Ss - 44%, 28% MNSs ANTIBODIES • Anti-M and Anti- N o Mostly ?? do not bind complement or react with enzyme treated cells. • Anti- M o Naturally occurring, saline reacting, IgM in nature o pH (6.0-6.5) & glucose dependent o Reaction is enhanced by anti-M lectin o Anti-M lectin - derived from Iberis amara • Anti- N o Cold reactive IgM and IgG saline agglutinin that does not bind complement or react with enzymes treated RBCs; not clinically significant unless reacts at 37C o pH independent o Seen in renal diseases (dialysis) ▪ Presence of anti-Nf o *Anti- N- formaldehyde induced anti- N o Anti-N lectin - derived from Vicea graminea, Bauhinia variegate, Bauhinia purpura Anti-M Anti-N Genotype Phenotype + MM M+N+ NN M-N+ + + MN M+N+ • • • • • • Consist of three antigens: ?? which are biochemically related to the CHO chain that makes up the ABH and I antigens Located on P1 antigen are P1 like Antigens o o Seen in P1 like Soluble Substances o Seen in o Seen in extracts from o Seen in extracts from P1 Antigens o Expression changes during fetal development (fetal RBC: 12 wks) o Already expressed starting 12th week and weakens with gestational age o Strength of P1 varies in race ▪ Blacks - strongest expression that whites o Easily/rapidly deteriorated during storage P ANTIBODIES • • • • Anti- P1 o Common, naturally occurring IgM ab in sera o Weak, cold-reactive, saline agglutinin reactive at 4C o ?? in the sera of P2 individuals o Usually ?? optimally reactive at o Can be NEUTRALIZED with Anti – P/Alloanti-P o Found naturally occurring in sera of all PK individuals o Hemolytic with a wide thermal range Autoanti- P (Donath Landsteiner antibody) Notes: o Biphasic – o Increased Temperature- • • • Anti- PP1k (Anti- Tja) o Separable thru adsorption o 1st seen in patient with o Associated with increased incidence on spontaneous abortions in early pregnancy o May demonstrate o Produced by all p individuals early in life without RBC sensitization Autoanti-P associated with paroxysmal Cold Hemoglobinuria o Cold reactive IgG autoantibody in PCH o Biphasic hemolysin o In vitro binds to RBCs in the cold and via complement activation once the coated RBCs are warmed to 37C Anti-PK o Selective adsorption with P1 cells o Can be inhibited with hydatid cyst fluid • • • KELL ANTIBODIES • I BLOOD GROUP SYSTEM (I: 027) • • • Found in chromosome 6 Discovered by Wiener “Individuality System” • • • • Antigens involved are seen in two different age groups: o i for neonates to 18 months o I for adults At birth, infant RBCs are rich in i; I is almost undetectable. During the 1st 18 months of life, the quantity of i slowly decreases as I increases until adult proportions are reached Adults red cells adult RBCs are rich in I and have only trace amounts of i antigen NOTE: There is no true I– or i– phenotype. The strength of I and i varies from individual to individual, and the relative amount detected will depend on the example of anti-I or anti-i used • • • I ANTIBODIES • • • Benign Anti – I o Weak naturally occurring saline reactive IgM autoagglutinin detectable only at 4C Pathologic Anti-I o Potent cold autoagglutinin that demonstrated high titer reactivity and reacts over a wide thermal range (0-30C) o Associated with Mycoplasma pneumoniae infection (Cold hemagglutinin disease) Anti- i o An IgM agglutinin and reacts optimally at 4C o Associated with infectious mononucleosis (IM) or kissing disease by Epstein Bahr Virus (EBV) • KELL BLOOD GROUP SYSTEM (Kel:006) • • • Found in chromosome 7 Named after Mrs. Kellaher/ Kelleher 2nd to D in terms immunogenicity KELL ANTIGENS • • • Are found only on red cells Rare antigen Well developed at birth & area not destroyed by enzymes Anti- K o Outside of ABO and Rh, anti- K is the most common antibody seen in blood bank o Usually IgG immune antibodies reactive in AHG phase o Can cause both HDN HTR o Most common antibody o Clinically significant PHENOTYPES I ANTIGENS • The K (Kell) antigen is 2nd rated only to D antigen in immunogenicity Destroyed or Inactivated by sulfhydryl reagent like AET, DTT, ZZAP (artificial Kell null) Notes: o AET - aminoethylisothiouronium bromide o DTT - Dithiothreitol o ZZAP - sulfhydryl reagent (dithiothreitol) and a proteolytic enzyme (papain or ficin) • Kx o Precursor Kell antigen o Responsible for the synthesis or formation of Kell ag o Found in the following cells: ▪ WBC • Precursor Kell is unchanged • Do not produce Kell • Kx(-): Chronic Granulomatous Disease (CGD) • Common among males ▪ RBC • Precursor Kell is converted/ changed to Kell • Kx (+): McLeod Syndrome KO or K null Phenotype o Lacks Kell antigens o No membrane abnormality McLeod Phenotype o Lacks the Kx antigen (which might be a precursor for Kell antigen) o Rare phenotype with decrease Kell system antigen expression and abnormal red cell morphology o McLeod Syndrome is associated with the following: ▪ Chronic but often well-compensated hemolytic anemia ▪ Reticulocytosis, Acanthocytosis (presence of irregular protrusions) ▪ Muscular dystrophy (↑ serum CK-MM/ CK1) ▪ Common among males suffering from Chronic Granulomatous Disease (CGD) K null vs. McLeod Phenotype K null McLeod Kx Antigen Present Lacking Abundant Autosomal Kell Lacking Decrease Antigen expression Red cell No Yes Abnormality Other Kell Antigens o Kell (K) o Rautenberg (Kpb) o Class (KL) o Cellano (k) o Sutter (Jsa) • o Williams (Kw) o Penny (Kpa) o Matthew (Jsb) o Peltz (Ku) Note: o Cellano (k) occurs in over 99% Caucasian & Negroes o Kell (K) occurs in 9% Caucasians and 3.5% Negroes LUTHERAN ANTIGENS • DUFFY BLOOD GROUP SYSTEM (Fy:008) • • • Named after hemophiliac patient, Mr. Duffy Clinically significant Destroyed by enzyme Lua and Lub o Poorly developed at birth o Lu (a-b-) – May result from three different genetic backgrounds: ▪ Dominant in (Lu) Types - expression of Lutheran was thought to be suppressed by a rare dominant gene, In (Lu) for “inhibitor of Lutheran” ▪ Recessive Lulu Type – lacks all Lu ag ▪ Recessive sex- linked inhibitor Type-X – borne inhibitor of Lutheran • Seen as result of Lutheran gene DUFFY ANTIGENS LUTHERAN ANTIBODIES • • Fya and Fyb o Well developed at birth o Long arm of chromosome 1 o Easily destroyed by common proteolytic enzymes o Were shown to resist infection caused by Plasmodium vivax and Plasmodium knowlesi o No cell membrane receptor for Plasmodium vivax and Plasmodium knowlesi merozoite (form of Plasmodium that is infective to RBCs; can readily penetrate/infect) • DUFFY ANTIBODIES • Anti-Fya and Anti-Fyb o Usually IgG and react best at the AHG phase o Both are implicated indelayed HTR (DHTR) and HDN KIDD BLOOD GROUP SYSTEM (Jk:009) • Named after Mrs. Kidd KIDD ANTIGENS • Jka and Jkb o Well developed at birth, contributing to potential HDN o Clinically significant o Serves as cell membrane channel for the entry of urea to the cells o Urea - used as lysing agent for red cells ▪ seen in trisomy 21 (Down syndrome) o Show in vitro hemolysis o Found on chromosome 18 o Reactivity enhanced by enzyme treatment KIDD ANTIBODIES • Anti- Jka and Anti- Jkb o Show dosage o Both are IgG immune antibodies (primarily IgG3 and anti-globulin) o Bind complement o Common cause of delayed hemolytic transfusion reaction LUTHERAN BLOOD GROUP SYSTEM (Lu:005) • • • • • • Lutheran genes is located on chromosome 19 Closely linked with secretor locus Misinterpretation of blood donor’s name No H gene Poorly developed at birth Seen in sialic acid glycoprotein of cell membrane Anti- Lua o Naturally occurring saline agglutinins that reacts best at room temperature (IgM) o Characteristically show loose and mixed fixed reactivity in vitro Anti- Lub o Most are IgG often IgG4 immune antibodies o Reactive at AHG at 37C and the AHG phase o Clinically significant MINOR BLOOD GROUPS DIEGO BLOOD GROUP SYSTEM (Di:010) • • • • • Antithetical Pair high incidence Dib/Wrb Antithetical Pair Low incidence Dia/Wrb Dia antigen is used for anthropological studies of Mongolian ancestry Dia and Dib is located on AE-1 also known as RBC band 3 responsible for the exchange of bicarbonate and chloride Mutations in AE-1 may result in hereditary spherocytosis, congenital acanthocytosis, South east Asian ovalocytosis CARTWRIGHT BLOOD GROUP SYSTEM (Yt:011) • • • Yta is a high incidence antigen while Ytb is a low incidence antigen It is found on then acetylcholinesterase that is responsible for neurotransmission Antibodies are mostly IgG Xg SYSTEM (Xg:012) • • • Located on the short arm of X chromosome 89% in female and 66% in male expresses Xga phenotype Associated with CD99 antigen produced by MIC 2 gene SCIANNA BLOOD GROUP SYSTEM (SC:013) • • • Antigens are resistant to most enzymes but sensitive to reducing agents Antigens are Sc1 and Sc2 Shown in RBC ERMAP that is expressed in erythropoiesis DOMBROCK BLOOD GROUP SYSTEM (DO:014) • • Associated with Doa and Dob antigen Linked to glycosylphosphatidylinositol glycoprotein associated with regulation of cellular protein fucntion COLTON BLOOD GROUP SYSTEM (CO:015) • • • • • Associated with Coa and Cob Located on Aquaporin-1 (AQP1) for regulation of ospotic water transport CHIDO/ ROGERS BLOOD GROUP SYSTEM (CH:017) • • • • • • Associated with CH1, CH2, CH3, RG1 and RG2 antigens Located on C4 complement component of RBC membrane Collectively grouped as HTLA (High Titer Low Avidity) HTLA means inconsistent reactions and often found with other antibodies. Titration and inhibition can be used for identification Can cause anaphylactic reaction in several cases GERBICH BLOOD GROUP SYSTEM (GE:020) • • Associated with Glycophorin C and D which is associated with RBC membrane band 4.1 Composed of GE2, GE3 and GE4 CROMER BLOOD GROUP SYSTEM (CROM;021) • • Sensitive to pronase treatment Carried by Decay Accelerating Factor (DAF) or CD55 that is involved in regulation of complement activation of C3 and C5 convertase KNOPS BLOOD GROUP (KN:22) • Associated with CR1 or CD25 that is responsible for immune adherence mechanism that delivers C3b complexes to the liver for removal INDIAN BLOOD GROUP (IN:023) • Associated with CD44 which is major component for T and B cell activation BENNETH GOODSPEED ANTIGEN • • • Bga, Bgb and Bg Directed toward HLA which are categorized as MHC class I and II Bga = HLA B-7, Bgb = HLA-B17, Bgc = HLA A-28 SUMMARY OF MAJOR AND MINOR BLOOD GROUPS IN SUMMARY • • • • • • • • • Le, P, I Lu, P1 - IgM naturally occurring antibodies (generally) but can become IgG K, Fy, Jk, Ss, Xga - IgG Immune Antibodies Anti-C/Anti-AB (ABO), Anti-f (Rh), Anti-U (MNSs U), Anti-D, Anti-Fya, Anti-K, Anti-Ss, Anti-Jk - Antibodies that causes HDN/EF Kidd, Rh, I, PI, Lewis - Enhance by Proteolytic Enzymes Duffy, MNSs U - Inactivated/ Destroyed by Proteolytic Enzymes Lewis, Lutheran, ABH - Blood groups associated with secretor genes MNSs U, Kidd, Kell - Antigens that are well developed at birth Lewis, P, I, Lutheran - Antigens that are poorly developed at birth Kidd, Duffy - Labile antigens M/N, S/s, K/k, Jka/Jkb, Rh ags expt. D - Antigen commonly showing dosage effect Kpa, Jsa, Lua - Low frequency antigens ABO, Rh, Kell, P, U, I, Kpb, Jsb, Lub - High Frequency Antigens CHROMOSOMAL LOCATIONS OF THE MAJOR AND MINOR BLOOD GROUPS CHROMOSOME LOCUS Major Blood Groups 1 Rh, Duffy 4 MNS 6 I 7 Kell 9 ABO 18 Kidd 19 Lewis, Lutheran, H 22 P1 Minor Blood Groups 1 Cromer, Scianna 2 Gerbich 11 Indian 12 Dombrock •