1 0 1 Neither A nor B I Both Anti-A and Anti-B 1 0, A, B, AB I 0 1. Simulated ABO 6; Rh Bood vping Lab Activity Student Study Guide BACKGROUND

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1 Smulated ABO 6; Rh Bood vpng Lab Actvty Student Study Gude BACKGROUND nces Around 900, Karl Landstener dscovered that there are at least four dfferent knds of human blood, determned by the presence or absence of specfc agglutnogens (antgens) on the surface of red blood cells (erythrocytes). These antgens have been desgnated as A and B. Antbodes aganst antgens A or B begn to buld up n the blood plasma shortly after brth, the levels peak at about eght to ten years of age, and the antbodes reman, n declnng amounts, throughout the rest of a person's of lfe. The stmulus for antbody producton s not clear; however, t has been proposed that antbody producton s ntated by mnute amounts of A and B antgens that may enter the body through food, bactera, or other means. Humans normally produce antbodes aganst those antgens that are not on ther erythrocytes: A person wth A antgens has ant-b antbodes; a person wth B antgens has ant-a antbodes; a person wth nether A nor B antgens has both ant-a and ant-b antbodes; and a person wth both A and B antgens has nether ant-a nor ant-b antbodes (Fgure ). Blood type s based on the antgens, not the antbodes, a person possesses. The four blood groups are types A, B, AB, and 0. Blood type 0, characterzed by the absence of A and B agglutnogens, s the most common n the Unted States and s found n 45% of the populaton. Type A s next n frequency, and s found n 39% of the populaton. The frequences at whch types B and AB occur are 2% and 4% respectvely. Fgure Antgens on Antbodes Can Gve Can Receve Erythrocytes n Plasma Type (Agglutnogens) (Agglutnns) Blood To Blood From B B Ant-A B, AB 0, B AB A and B Nether Ant-A nor Ant-B AB 0, A, B, AB 0 Nether A nor B Both Ant-A and Ant-B 0, A, B, AB 0 Copymaster. Permss~on granlcd lo makc unlmted copes Tor usc n any one school buldng. For educalonal use only. Not Tor commercal usc or resale. O 2002 WARD'S Natural Scence Establshment All Rghts Reserved

2 ABO System Process of Agglutnaton There s a smple test performed wth antsera contanng hgh levels of ant-a and ant-b agglutnns to determne blood type. Several drops of each knd of antserum are added to separate samples of blood. f agglutnaton (clumpng) occurs only n the suspenson to whch the ant-a serum was added, the blood type s A. f agglutnaton occurs only n the ant-b mxture, the blood type s B. Agglutnaton n both samples ndcates that the blood type s AB. The absence of agglutnaton n any sample ndcates that the blood type s 0 Agglutnaton Reacton of ABO Blood-Typng Sera Reacton Ant-A Serum Ant-B Serum Agglutnaton No Agglutnaton No Agglutnaton No Agglutnaton No Agglutnaton mportance of Blood Typng DD YOU KNOW? Donor blood contabs only As noted n the table above, people can receve transfusons of only certan blood types, dependng on the type of blood they have. f ncompatble blood types are mxed, erythrocyte destructon, agglutnaton and other problems can occur. For nstance, f a person wth type B blood s transfused wth blood type A, the recpent's ant-a antbodes wll attack the ncomng type A erythrocytes. The type A erythrocytes wll be agglutnated, and hemoglobn wll be released nto the plasma. n addton, ncomng ant-b antbodes of the type A blood may also attack the type B erythrocytes of the recpent, wth smlar results. Ths problem may not be serous, unless a large amount of blood s transfused. no antbodes red blood cells are often used n medco-legal stuatons nvolvng dentfcaton of dsputed paternty. A comparson of the blood groups of mother, chld, and alleged father may exclude the man as a possble parent. Blood typng cannot prove that an ndvdual s the father of a chld; t merely ndcates whether or not he possbly could be. For example, a chld wth a blood type of AB, whose mother s type A, could not have a man whose blood type s 0 as a father. Copymaster. Pamsson grankd to make unlmted copes for use n any om 2 school buldng. For educatonal use only. No for commercal use or resale WARD'S Natural Scence Establshment All Rghts Reserved

3 F-"-- S -v--w-?-*---<--p--m-< The Genetcs of Blood Types B f The human blood types (A, B, AB, and 0) are nherted by multple alleles, whch occurs when three or more genes occupy a sngle locus on a chromosome. Gene A codes for the synthess of antgen (agglutnogen) A, gene B codes for the producton of antgen B on the red blood cells, and gene does not produce any antgens. The phenotypes lsted n the table below are produced by the combnatons of the three dfferent alleles: A, B, and. When genes B and A are present n an ndvdual, both are fully expressed. Both A and B are DD YOU KNOW? domnant over so the genotype of an ndvdual wth blood type 0 Camels and ther relatves are be (Fgure 3). the only mammals havng oval red blood cells. Fgure 3 Use A for antgen A, 6 for antgen 8, and for no antgens present. Genes A and B are domnant over. AB blood type results when both genes A and B are present. Rh System n the perod between 900 and 940, a great deal of research was done to dscover the presence of other antgens n human red blood cells. n 940, Landstener and Wener reported that rabbt sera con- 0 tanng antbodes for the red blood cells of the Rhesus monkey,\' '+ would agglutnate the red blood cells of 5% of Caucasans. These antgens, sx n all, were desgnated as the Rh (Rhesus) factor, and they were gven the letters C, c, D, d, E, and e by Fscher and Race. Of! o0 these sx antgens, the D factor s found n 85% of Caucasans, 94% of DD YOU KNOW? Afrcan Amercans, and 99% of Asans. An ndvdual who possesses ( ~h s so named because the these antgens s desgnated Rh+; an ndvdual who lacks them s ntal study was done wth desgnated m- $ Rhesus monkeys. Phenotype A Possble Genotypes B AB 0 The genetcs of the Rh blood group system s complcated by the fact that more than one antgen can be dentfed by the presence of a gven Rh gene. ntally, the Rh phenotype was thought to be determned by a sngle par of alleles. However, there are at least eght alleles for the Rh factor. To smplfy matters, consder one allele: Rh+ s domnant over Rh-; therefore, a person wth an Rh+/Rh- or Rh+/Rh+ genotype has Rh+ blood. 3-" - r r r " * ~ ~ ~ - - r " * - ~ - ~ x > ~ r. r - ~ rpa-*runau^ '"rxa* xx- u x W -. - ~ ~ ~ ~ * - * - - ~ ~ AB - Copymaster. Permsson granted Lo makc unlmted copcs for ur n any one school buldng. Fw educatonal use only Not for commercal usc or resale. 3 O 2002 WARD'S Natural Scence Establshment All Rghts Reserved

4 The ant-rh antbodes of the system are not normally present n the plasma, but ant-rh antbodes can be produced upon exposure and senstzaton to Rh antgens. Senstzaton can occur when Rh+ blood s transfused nto an Rh- recpent, or when an Rh- mother carres a fetus who s Rh+. n the latter case, some of the fetal Rh antgens may enter the mother's crculaton and senstze her so that she begns to produce ant-rh antbodes aganst the fetal antgens. n most cases, senstzaton to the Rh antgens takes place toward the end of pregnancy, but because t takes some tme to buld up the ant-rh antbodes, the frst Rh+ chld carred by a prevously unsenstzed mother s usually unaffected. However, f an Rh- mother, or a mother prevously senstzed by a blood transfuson or a prevous Rh+ pregnancy, carres an Rh+ fetus, maternal ant-rh antbodes may enter the fetus' crculaton, causng the agglutnaton and hemolyss of fetal erythrocytes and resultng n a condton known as erythroblastoss fetals (hemolytc dsease of the newborn). To treat an nfant n a severe case, the nfant's Rh+ blood s removed and replaced wth Rh- blood from an unsenstzed donor to reduce the level of ant-rh antbodes. Blood Components DD YOU KNOW? Erythrocytes are crcular, bconcave dsks of 5 to 8 mcrometers. Ther Leukocytes are capable of chef functon s to transport oxygen (02) and carbon doxde (CO2). amoebod movement and are The transport of 0 2 and COz depends largely on the hemoglobn presometmes called amoebo- sent n the erythrocytes. The bconcave shape s also related to the erythrocytes functon of transportng gases, n that t provdes an ncreased surface area through whch gases can dffuse. The number of crculatng RBCs s closely related to the blood's oxygen-carryng capacty. Any changes n the RBC count may be sgnfcant. RBC counts are routnely made to dagnose and evaluate the course of varous dseases. Leukocytes range n sze from approxmately 9 to 25 mcrometers and functon prmarly to control varous dsease condtons. Leukocytes can move aganst the current of the bloodstream through amoebod movement, and pass through the blood vessel walls to enter the tssues. The total WBC count normally vares from 5,000 to lo,ooo/mm3. Certan nfectous dseases are accompaned by an ncrease n WBCs. f the number exceeds lo,ooo/mm3, the person has an acute nfecton. f t drops below 5,00O/mm3, the person may have a condton such as measles or chcken pox. The percentage of the dfferent types of leukocytes present n the blood may also change n partcular dseases, ths number s mportant for dagnostc purposes and s called a dfferental count. Copymaster. Permsson granted lo make unlmted copcs Tor use n any one 4 school buldng. For cducalonal usc only. Not Tor commercal use or rcsale. O 2002 WARD'S Natural Scence Establshment All Rghts Resewed

5 Defne agglutnogen and agglutnn Perform an actual blood typng procedure Observe the antgen/antbody reacton n smulated blood Determne the ABO and Rh blood type of four unknown normal blood Understand requrements for blood transfusons MATERALS MATERALS NEEDED PER GROUP D YOU KNOW? 4 Blood typng sldes 2 Toothpcks Mcroscope slde Coverslp SHARED MATERALS 4 Unknown blood samples: Mr. Green Ms. Brown Smulated Ant-A Serum Smulated Ant-B Serum Smulated Ant-Rh Serum PROCEDURE Although WARD'S Smulated Blood s completely safe, nonbologcal, and non-toxc, you should wear the proper personal protectve equpment to mmc the experence of an actual hematology laboratory. PART A: ABO and Rh BLOOD TYPNG. Label each blood typng slde: Slde #: Mr. Smth Slde #2: Mr. Jones Slde #3: Mr. Green Copymaster. Permsson granted to make unlmled copes ror use n any one 5 school buldng. For educatonal use only. Not Tor commercal use or resale. O 2002 WARD'S Natural Scence Establshment All Rghts Reserved

6 2. Place three to four drops of Mr. Smth's blood n each of the A, B, and Rh wells of Slde #. 3. Place three to four drops of Mr. Jones's blood n each of the A, B, and Rh wells of Slde #2. 4. Place three to four drops of Mr. Green's blood n each of the A, B, and Rh wells of Slde #3. 5. Place three to four drops of Ms. Brown's blood n each of the A, B, and Rh wells of Slde #4. 6. Place three to four drops of the smulated ant-a serum n each A well on the four sldes. 7. Place three to four drops of the smulated ant-b serum n each B well on the four sldes. 8. Place three to four drops of the smulated ant-rh serum n each Rh well on the four sldes. 0. Observe each slde and record your observatons n Table of the Analyss secton. To confrm agglutnaton try readng text through the mxed sample. f you cannot read the text, assume you have a postve agglutnaton reacton. Agglutnaton No Agglutnaton. Dspose of all materals accordng to your teacher's nstructons. WARD'S Sm4lated blood s non-bologcal and nontoxc and may be flushed down the dran. Be sure to wash and save the blood typng trays and toothpcks for future use. Copymaster. Pmsson pnbd lo make unlmlcd copes Tor use n any one schwl buldng. For educalonal use only. Not lor commercal use or resale. O 2002 WARD'S Natural Scence Establshment 6 All Rghts Reserved

7 PART B: Blood Cell Count. Thoroughly shake one of the vals of WARD'S Smulated Blood. Add one drop of smulated blood to a mcroscope slde, and cover wth a coverslp. Lower the coverslp slowly to avod trappng ar bubbles on the slde. 2. Examne the slde wth the low power (0X). Fnd an area of the slde wth an even dstrbuton of cells. 3. Swtch to hgh power (40X). Refocus and count the number of smulated red blood cells (red spheres) n the feld of vew. Count the cells n any clump separately. Record the number n Table Count the number of smulated whte blood cells (blue spheres). Record the number n Table Repeat the countng procedure wth two other felds of vew. Record these counts n Table Calculate the average of the three red blood cell counts and the f three whte blood cell counts. Record the results n Table Multply the average number of red and whte blood cells by the dluton factor to determne the number of red and whte blood cells per cubc mllmeter. Record each value n Table Dspose of all materals accordng to your teacher's nstructons. WARD'S Smulated blood s non-bologcal and nontoxc and may beflushed down the dran. Copymuster. Permlsslon pnled lo make unllmnled coples for uuse n any one xhool bu~ld~ng For educat~onal use only Not for commercal usc or resale WARD'S Natural Scence Establshment All Rghts Reserved

8 ANALYSS Table Ant-A Serum Ant-B Serum Ant-Rh Serum Blood Type Slde # Mr. Smth Slde #3 Mr. Green Slde #4 Ms. Brown Table 2 ~lood cell Count Total # Avg. # Cells Dluton Total # Blood Cells per mm3 or Type 2 3 of cells or Total3 Factor Avg. # Cells x Dluton Factor 50,000 5,000 Copymaster. Pcrmsslon granlcd to make unlmted copes br ur n any one school buldng. For cducalonal use only. No for commercal use or resale. O 2002 WARD'S Natural Scence Establshment All Rghts Reserved