AMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY 73:l-7 (1987) Blood Types of the Native Americans of Oklahoma D.O. KASPRISIN, M. CROW, C. McCLINTOCK, AND J. LAWSON American Red Cross Blood Seruices, Tulsa, Oklahoma 74128 KEY WORDS Blood groups, Gene frequencies, Cherokee, Creek, Choctaw, Diego blood group system ABSTRACT Large numbers of Indians from Oklahoma were screened for a variety of red cell antigens. Sufficient numbers of Cherokees, Creeks, and Choctaws were studied to calculate gene frequencies. These tribes originated in the Southeastern United States and were forcibly moved to Oklahoma. The Creeks and Choctaws have not been studied previously. A small number of Cherokees remained in North Carolina, and their blood types have been reported. The blood types of the Oklahoma Cherokees are quite similar to those observed there but one important difference was discovered. The data previously reported concerning the Eastern Cherokees revealed the absence of the Dia antigen. The present study found that the Oklahoma Cherokees do have the Dia antigen, although in a lower percentage than the other southeastern tribes. The Creeks and Choctaws share a linguistic heritage as well as having similar red cell phenotypes. The state of Oklahoma was designated as Indian Territory by the federal government, and throughout the 19th century many tribes of Native Americans were forcibly moved to this state from their homelands. The history of the tribes living in Oklahoma has been described by Wright (1951). Today, 67 tribes are represented in this state, including several small tribes and others which have merged in the distant past and have lost their individual identities. It is estimated that 180,000 Native Americans now reside in Oklahoma. The first tribes moved to this state were the Cherokee, Creek, Choctaw, Chickasaw, and Seminole, collectively known as the Five Civilized Tribes, a term originated by the Office of Indian Affairs because of their early and remarkable advancement. While most of the Cherokee tribe was moved to Oklahoma, a few hundred hid from the U.S. Army and remained in their original homeland. Their descendants live on the Qualla reservation in western North Carolina. The blood types of these eastern Cherokee were studied by Snyder (1926), Pollitzer et al. (19621, and Spees et al. (1975). However, since these are descendants of a small portion of the tribe, they may not be truly representative of the tribe as a whole. The blood types of the Cherokee of Oklahoma and many 0 1987 ALAN R. LISS, INC. other Oklahoman tribes have not been studied. This report examines the blood types of the Choctaw, Creek, and Cherokee, which are among many Oklahoma tribes not previously studied. The results obtained from the Cherokee of Oklahoma are compared to the previous studies of the eastern Cherokee. MATERIALS AND METHODS Beginning in 1982, the American Red Cross Blood Services, Oklahoma Region, began a search for rare blood donors among the Native American blood donors. The blood center region encompasses the entire northeast, and part of the southeast and north central, portions of Oklahoma. Large numbers of Cherokee, Choctaw, and Creek reside in this area. Figure 1 illustrates the tribal locations in Oklahoma and demarcates the blood center region. The method for attracting the participation of Native Americans in this project has been published previously by McClintock et al. (1984). Endorsement from the Principal Chiefs of the Cherokee and Creek Nations was obtained and a poster prepared for all blood collection sites which explained the need for donors with particular rare blood Received August 11,1986;revision accepted December 1,1986. Fig. 1. Tribal locations in Oklahoma. The shaded area demarcates the blood center region. (Reproduced with permission from Historical Atlas of Oklahoma, Second Edition, Revised and Enlarged, by John W. Morris, Charles R. Gains, and Edwin C. McReynolds. Copyright 1965, 1976 by the University of Oklahoma Press.) 3 BLOOD TYPES OF NATIVE AMERICANS OF OKLAHOMA types and how ready availability of these rare types would be beneficial for those Native Americans needing them. Blood center personnel were present to answer inquiries. All blood donors who volunteered to participate were asked to complete a consent form which detailed the donor’s percentage of Native American ancestry and tribal identification. More than 4,000 Native American blood donors representing 30 tribes volunteered to list their heritage. The largest number of participants were from the Cherokee, Choctaw, and Creek tribes. The location of the Seminole and Chickasaw tribes is south of the blood center region, and few members of these tribes were recruited. All participants were screened for the Dia antigen. Because of the rarity of the Dib antisera, only those Native Americans who were positive for Dia were further studied for Dib. Blood donors who claimed to be more than three-quarters Indian had a more complete red cell typing. The ABO, Rh, MNSs, D a y , Kidd, Colton, Kell, and Diego systems were studied. Three hundred samples from donors three-quarters or more Indian were also tested for Wra, He, Vw, Js”, Mg, and Kpa. No positives were found, and no further study of these antigens was conducted. Anti-Dia was provided by the American Red Cross National Headquarters. All Di(a+) samples were also tested with a commercial anti-Dia. Anti-Dib was provided by SCARF (Serum, cells, and rare fluids, an international immunohematological exchange group). All samples that were Di(a-) were considered Di(a-b+) since neither a third allele in this system nor the Di(a-b-) phenotype has been identified. Antigen testing in this study was done by microplate technique. Positive tests for lowincidence antigens, i.e., Cob,Dia, and K, were tested with the tube test and repeated with antisera from at least one other source. Gene frequencies of the MNSs and Rh systems were computed by Henry Gershowitz, Ph.D., at the University of Michigan Medical School by the maximum-likelihood method. The ABO gene frequencies were calculated by the Bernstein (1930) method. All other blood group systems were calculated by simple gene counting. RESULTS Tables 1-5 present the red cell phenotypes and gene frequencies found in the Cherokee, Creek, and Choctaw tribes. Many of the individuals studied were from other tribes whose total number of participants was not sufficient to list separately. Many others claimed mixed Indian ancestry. This group is listed in the tables as other tribes and mixed ancestry and acts as a group of Indians from various tribes for comparison. Approximately 31% of this group were full-blood Indians from tribes whose numbers were too small to permit statistical analysis; 50%were full-blood Indians who claimed more than one tribe, and 18% claimed some Caucasian or other non-Indian ancestry. Only those participants who claimed more than three-quarters Indian blood are included in these tables. Of the more than 900 Indians studied, the largest number (>250) were from the Cherokee tribe. More than 150 Creek and 60 Choctaw are included. Not all blood samples could be tested for all antigens and the number of participants varies. TABLE 1. ABOphenotypes of the Indians of Oklahoma ABO phenotypes (47,) ABO phenotype A B 0 AB Total tested Gene frequencies A B 0 Choctaw Other tribes and mixed ancestry 154 7 (10) 0 (0) 62 (90) 0 (0) 69 152 (29) 12 (2) 356 (68) 1(< 1) 521 0.0595 0.0388 0.9017 0.0521 0.0000 0.9479 0.1243 0.0205 0.8552 Cherokee Creek 52 (19)* 15 (6) 197 (74) 3 (1) 267 17 (11) 11 (7) 125 (81) 1(1) 0.1069 0.0322 0.8609 *Percentages have been rounded and may not total 1006. 4 D.O. KASPRISIN ET AL. TABLE 2. Rh phenotypes and gene frequencies of the Indians of Oklahoma Rh phenotype (%) Cherokee Probable Rh phenotype RlR1 R1r RlR2 R2Rz Rzr RZRl RzRz RzRz Ror rr r‘r r”r Total tested Gene freauencies Rl * R2 r Ro Rz r’ Creek 73 (27)* 52 (19) 72 (27) 18 (7) 21 (8) 16 16) 9 (3) 1(<1) 1(<1) 4 (2) 0 (0) 0 (0) 267 56 (36) 18 (12) 35 (23) 17 (11) 10 (6) 11(7) 6 (4) 1(1) 0 (0) 0 (0) 0 (0) 12 (17) 7 (101 25 (36) 7 (10) 11(16) 0 (0) 154 0 (0) 69 0.5261 0.2490 0.1475 0.5670 0.2715 0.4187 0.3752 > R, or r 0.0954 0.1537 0.0662 0.0000 0.0000 0.0523 0.0000 0.0000 0.0174 0.0600 0.0000 0.0000 r“ Choctaw Other tribes and mixed ancestry 0.5009 0.3163 0.1116 0.0191 0.0354 0.0076 0.0098 *Percentages have been rounded and may not total 1006. TABLE 3. MNSs phenotypes and gene frequencies of the Indians o f Oklahoma MNSs phenotypes (%) Cherokee Creek Chocktaw Other tribes and mixed ancestry MNSs phenotype MMSs MMss MMSS MNSs MNss MNSS NNSs NNss NNSS Total tested Gene frequencies MS Ms NS Ns 70 126)* 50 (19) 11(4) 59 (22) 47 (18) 12 (5) 7 (3) 8 (3) 2 (1) 266 36 (24) 25 (16) 16 (11) 29 (19) 25 (16) 0.2528 0.4615 0.0969 0.1888 0.3142 0.3949 0.0845 0.2064 155 (30) 117 (22) 46 (9) 69 (13) 83 (16) 18 (3) 12 (2) 21 (4) 4 (1) 524 0.2686 0.5502 0.0937 0.0875 0.2946 0.4735 0.0594 0.1725 *Percentages have been rounded and may not total 100%. The phenotypes and gene frequencies are similar to those found in other studies of Indian populations. The majority of all four groups of Native Americans were group 0, the highest percentage occurring in the Choctaw tribe. The percentage of group B was very low and was absent in the Choctaw. Elevated frequencies of R1, R2, and R, were found, and the frequency of r was correspondingly low. R,, r’, and r ” were very rare and r y was not found. The frequency of R, and r were so low in the Creek and Choctaw tribes 5 BLOOD TYPES OF NATIVE AMERICANS OF OKLAHOMA TABLE 4. Duffy and Kidd phenotypes and gene frequencies of the Indians of Oklahoma Duffy phenotypes Fy(a +b-) Fy(a-b+) Fy(a+b+) Total tested Gene frequencies FYa FYb Kidd phenotypes Jk(a +b -1 Jk(a -b +) Jk(a +b +) Total tested Gene frequencies Jk" Jkb Cherokee (%) Creek (%I Choctaw (%) Other tribes and mixed ancestry (%) 77 (29)" 38 (14) 149 (56) 264 60 (39) 22 (14) 72 (47) 154 25 (36) 9 (13) 35 (51) 69 218 (42) 53 (10) 245 (48) 516 0.5739 0.4261 0.6234 0.3766 0.6159 0.3841 0.6599 0.3401 43 (16) 57 (22) 162 (62) 262 28 (18) 36 (24) 89 (58) 153 6 (9) 13 (19) 50 (72) 69 135 (26) 102 (20) 279 (54) 516 0.4733 0.5267 0.4739 0.5261 0.4493 0.5507 0.5320 0.4680 *Percentages have been rounded and may not total 100%. TABLE 5. Kell, Colton, and Diego phentotypes and gene frequencies Kell phenofypes K+k+ K-k+ Total tested Gene frequencies K k Colton phenotypes Co(a+b-) Co(a +b +)3 Total tested Gene frequencies coa Cob Diego phenotypes Di(a +b-) Di(a-b+)4 Di(a+b+) Total tested Gene frequencies Di" Dib Other tribes and mixed ancestrv (%) Cherokee (%) Creek (%) Choctaw (%I 6(212 258 (98) 264 2 (1) 152 (99) 154 3 (4) 66 (96) 69 16 (3) 500 (97) 516 0.0114 0.9886 0.0065 0.9935 0.0217 0.9783 0.0155 0.9845 241 (97) 7 (3) 248 104 (96) 4 (4) 108 66 (96) 3 (4) 69 490 (98) 9 (2) 499 0.9859 0.0141 0.9815 0.0185 0.9783 0.0217 0.9910 0.0090 0 (0) 255 (96) 12 (4) 267 1(1) 135 (88) 17 (11) 153 0 (0) 63 (91) 6 (9) 69 3 (1) 482 (93) 8 (7) 523 0.0225 0.9775 0.0621 0.9379 0.0435 0.9565 0.0421 0.9579 'k was not tested in K- subjects but assumed to be present. 'Percentages have been rounded and may not total 100%. 'Cod was not tested in Coh-) subjects but assumed to be present 4Dibwas not tested in Di(a-) subjects but assumed to be present. that the computerized analysis of gene frequencies could not separate these two genes. Only 2% of Cherokees and the mixed group were Rh negative. No Rh-negative donors were found among the 177 Creek and Choctaw studied. The frequency of M, S, Fya, and Jka was higher than that of the Caucasian population and similar to that of other Native Americans studied. None of the first 300 donors studied were positive for the Wr", He, V", Jsa, Mg, and Kpa antigens. The Dia antigen was present 6 D.O. KASPRISIN ET AL. in all four groups of Native Americans. The highest percentage was among the Creeks. Unlike the study by Pollitzer et al. (1962), which concluded that the Dia antigen was not present in the eastern Cherokees, this study found significant numbers of Di(a+) individuals among Oklahoma Cherokees, although less than the other groups in this study. DISCUSSION The Dia antigen is found almost exclusively among the Mongoloid race. The highest rate is found in South and Central American Indians, although there is considerable variation within a region. The rate decreases among Indians the further north one studies and is virtually absent in Alaska and Western Canada. Many theories have been proposed t o explain the origin of the American Indian and the gene frequencies reported in the various tribes. Whether the variability in Native Americans is due to diversification in a single slowly expanding group through genetic drift and natural selection or due t o multiple migrations is unknown. It has been hypothesized by Mourant et al. (1976)that the Dia antigen is absent in the earliest arrivals to this hemisphere. Therefore, if the antigen is absent in a population, that tribe may have arrived prior t o carriers of the gene. Tills (1982)suggests that there may have been as many as five migra- tions of Indians in the Americas with varying percentages of Dia genes in these populations. The recent review by Szathmary (1984) examines the question of a single migration or multiple migrations from Asia to America and is of particular interest. This study documents the existence of the Dia antigen among the tribes that originated in the Southeastern United States. The Cherokee Indians are part of the Iroquoian linguistic group, while the Creek and Choctaw tribes belong to the Muskhogean linguistic family. Linguistic similarity does not necessarily imply similar genetic backgrounds. Although this study found several differences in gene frequencies of red cell antigens between the eastern and Oklahoma Cherokees, the two groups are quite similar. Table 6 compares the gene frequencies for each blood group system studied in this project to the Snyder (1926) and Pollitzer et al. (1962) studies. Since the majority of the tribe was moved to Oklahoma, the small number of Cherokees that remained in North Carolina, a relatively inbred population, may not be truly representative of the Cherokee people. Pollitzer et al. (1962) were surprised at the absence of the Dia antigen in the eastern Cherokees. They noted that the Iroquois of New York State, who are part of the same linguistic group, demonstrated the Di(a+) phenotype in 10% of the members studied. The percentage of Dia is relatively low in the TABLE 6. Comparison of gene frequencies in Eastern Cherokees with Oklahoma Cherokees Gene 0 A B R1 R2 Rz R, or r r', r", r y MS Ms NS Ns K k Jk" Jkb FYa FY" Dia Oklahoma Cherokees: % or more Eastern Cherokees: full bloods Eastern Cherokees: mixed 0.8609 0.1069 0.0322 0.5261 0.2490 0.0600 0.1649 0.0000 0.2528 0.4615 0.0969 0.1888 0.0114 0.9886 0.4733 0.5267 0.5739 0.4261 0.0225 0.9727 0.0182 0.0091 0.6154 0.2885 0.0577 0.0385 0.0000 0.3694 0.4319 0.0793 0.1194 0.0064 0.9936 0.4694 0.5306 0.5471 0.4529 0.0000 0.8257 0.1399 0.0344 0.6702 0.2447 0.0213 0.0638 0.0000 0.2444 0.4418 0.0322 0.2816 0.0053 0.9946 0.4574 0.5426 0.6580 0.3420 0.0000 BLOOD TYPES OF NATIVE AMERICANS OF OKLAHOMA Oklahoma Cherokee, and sample selection could easily account for the difference with the eastern Cherokees. In addition, differing patterns of intermarriage could also have contributed to the differences. The Cherokees in this study were also found to have a higher percentage of A, N, s, and r. In the study by Spees et al. (1975), they studied Cherokees from North Carolina who were at least five-eighths Indian. They found similar gene frequencies for the Rh system m1=0.6613; R2=.2339; R,=0.0564; Ro or r=0.0484). We calculated the ABO gene frequencies by the Bernstein method based on the phenotype data published in this paper (A=0.1073; B =0.0050; 0 =0.8877). The Cherokees originated in the upper Ohio River valley south of the Iroquois tribe. The Iroquois and Cherokee tribes were bitter enemies, and the Cherokees were pushed further south where they came into contact with the Creeks and other tribes. Frequent struggles broke out between the Creeks and Cherokees during the 18th century. Prisoners captured during wars were occasionally allowed to assimilate into the capturing tribe. Because of the close proximity of the Cherokees with tribes with relatively high percentages of the Dia gene in their original home as well as in Oklahoma, this gene may have been introduced relatively recently if it was not already present. Noting the similarities and differences found among the Creek, Choctaw, and Cherokee tribes, further delineation of other genetic markers would be of interest to help clarify the origins and genetic interrelationship of these tribes. The blood center is now examining the HLA types of the Cherokee, Creek, and Choctaw tribes. 7 ACKNOWLEDGMENTS The authors are grateful to Henry Gershowitz, Ph.D., of the University of Michigan Medical School, for calculating the gene frequencies of the Rh and MNSs systems. We also wish to acknowledge the help of the many Native American leaders who endorsed and supported this work and the thousands of Native American blood donors for their participation in this study and their selfless contributions as blood donors. LITERATURE CITED Bernstein, F. (1930) Fortgesetzte Untersuchungen aus der Theorie der Blutgruppen. Z. Indukt. Abstramm. VererbLehre 56t233-272. McClintock, C, Crow, M, Lawson, J, and Kasprisin, DO (1984)The search for rare blood donors among Native Americans. Med. Lab. Observer 16(10):70-74. Morris, JW,Goins CR, and McReynolds EC (1976) Historical Atlas of Oklahoma. 2nd Edition. Norman, OK: University of Oklahoma Press. Mourant, AE, Kopec AC, and Domaniewska-Sobczak, K (1976) The Distribution of Human Blood Groups and Other Polymorphisms. London: Oxford University Press, pp. 21, 117-122. Pollitzer, WS, Hartmann, RC, Moore, H, Rosenfield, RE, Smith, H, Hakim, S, Schmidt, PJ, and Leyshon, WC (1962) Blood types of the Cherokee Indians. Am. J. Phys. Anthropol. 20t33-43. Snyder, LH (1926) Human blood groups: Their inheritance and racial significance. Am. J. Phys. Anthropol. 9t233-263. Spees, EK, Woodbury, MA, Gottsch, LG, Nicely, PR, and Amos, DB (1975) Heterogeneity of HL-A specificities in the Cherokee Indians. In: Kissmeyer-Nielsen, F. (ed): Histocompatibility Testing 1975. Copenhagen: Munksgaard, pp. 219-224. Szathmary, E J E (1984) Peopling of northern North America: Clues from genetic studies. Acta Anthropogenet. 8:79-109. Tills, D (1982) Blood groups of the Americans. Biotest Bull. 3:212-221. Wright, MH (1951)A Guide to the Indian Tribes of Oklahoma. Norman, OK: University of Oklahoma Press, pp. 1-300.