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HLA-A2 alleles in Northeast-Asian populations
50
Abstracts
11.1. Anthropology Today: Contribution of HLA (I) 0258
0257
DNA ITPING OF HLA CLASS [ GENES REVEALS FURTHER GENETIC DIFFERENCE BE1WEEN NORl'HERN AND SOUTHERN CHINESE
HIGH RESOLUTION MOLECULAR TYPING OF HLA-A AND HLA-B IN THREE SOUTH AMERICAN INDIAN TRIBES
Xiaofang Sun, YipingSun. Xiaojiang Gao Guangzhou Second People's Hospital, Guangzhou, China, The Australian National University, Canberra, Australia
Fernandez-Vitia MA, Lazaro AM, Marcos YC, NulfCJ, Fish VM, McGarry JE, Raimondi EH, Stastny P, Dept oflnternal Medicine, UT Southwestern Medical Ctr, Dallas, TX, USA
Our previous serological and class II DNAanalyses have demonstrated markedly different HLA profiles in the northern and southern Chinese.
We analyzed 187 unrelated individuals from the Toba (TOB, n=122), Wichi (WlC, n=47) and Pilaga (pIL n~18) tribes from NE Argentina. The number of alleles at the HLA-A locus was small (TOB n= 6, WlC n=6 and PIL n=4). In contrast, the HLA-B locus presented larger allelic variation (TOB n=23, WlC n=13, PIL n~14). HLA-A alleles consisted of A*0201, 2402,2403,68012,3101. A putative new allele, locally called A-02X09, with a hybridization pattern similar to A*020 I, but differing in codons 142167 in which it is similarto A*2402, was observed in TOB and WlC. All HLA-A locus alleles had similar gene frequencies which resulted in low homozygosity frequencies (hgfTOB= 0.18, WlC= 0.20, PIL=0.25). In TOB we observed two subtypes ofHLA- B5, three ofB35, 6 ofBl5, three ofB48, three ofB39 and 5 ofB40. We observed several new hybridization patterns which appeared to correspond to new alleles. Among them was a new subtype ofB48 which was the most frequent B-locus allele in this population (gf=0.15). In these inbred populations, in which polymorphism ofHLA-A and 8 loci is restricted, balanced allelic frequencies at both Class I loci appear to have resulted from selection for heterozygosity. However, a larger polymorphism and more frequent generation of new aUeles at the HLA-B locus. suggests that different evolutionary forces may have acted on each of these HLA Class I loci.
In the present experiments using comprehensive DNA typing protocols
developed in our laboratory we examined the HLA-A and -B genes in 85 northern (Beijing) and 160 southern [Guangdong) Chinese donors for serologically identical subtypes. Several antigens showed significantly different distrtbutions including Al, A26, A30, A3!. B37. B44. B48, B52 and B61 predominantly found in the north and All. B38. B46, B55, and B58 predominantly or only detected in the south. A2 the only A antigen further split by DNA typing showed extraordinary heterogeneity with six subtypes detected including A"0201, 0203, 0205, 0206. 0207 and 0210. In the B locus, however, 1/3 of the serological specificities were found to have hidden subtypes. Further, several antigens commonly seen in
both groups were found to be different DNAsubtypes including A"0201, B'0702, 1302, 1501, 2705, and 4006 in the north and A'0203, 0207, 8'0705, 130!. 1502, 2704 and 4001 in the south. In accordance with our previous reports these results reflect historical population mixtures with different genetic inputs in northern and southern China. The
previously
undetectable
hidden
subtypes
may have
important
implications for unrelated-donor transplantations in these people.
0259 HLA
CLASS
I
AND
CLASS
II
ALLELES
IN
THREE
SYMPATRIC WEST AFRICAN ETHNIC GROUPS Gaia LuonP·2, Maria De Luca1 ,3, David Modiano-, Mario Coluzzl'', Steven
GEMarsh! & JuliaG Boomer"'Tissue Antigen Laboratory, Imperial Cancer Research Fund,London, UK 21nstitute of Parasilology, University "LaSapienza", Roma, Italy 'Dept. ofCellular Biology. University ofCalabria, Arcavacala di Rende, Ilaly We havecollected a totalof 463samples from three West African populations: 187 Massi, 160 Rimaibe and 116 Fulani. These three elhnicgroupsare sympatric and have totally diflerent genelicand cultural backgrounds; while Mossi and Birnaioe are a Sudanese Negroid sedentary population, the Fulaniare descended from nomadic pasioralists and have non-Negroid features of possibleCaucasianorigin. Inorder to investigate the past and present genetic relationships betweenthese Ihree populations we are typing these samples [or HLA Class I using the 12th Workshop SSP ARMS-PCR typing kit and for HLA Class II using the 12th Workshop SSG-PCRtechnique. Preliminary data indicates a highfrequency of Cw·1701. Cw'04. B'5301 and B'4201 alleles in the Mossi tribe. Data will be presented on the allele and haplotype frequencies of all three populations, together with comparisons of genetic distance to other African and European populations,
0261
HLA class I polymorphism in Inuits.
0260 HLA-A2ALLELES IN NORTHEAST-ASIAN POPULATIONS Yoshihide Ishikawa', Katsushi Tokunaga'r, Jean-Marie Tiercy', Kouichi Kashiwase', Hidenori Tanaka', Jie Liu', Tatsuya Akaza', Kenji Tadokoro', Nyarn-Osorin Chimge",Guan JIUI Jia', Takeo Juji' I. Japanese Red Cross Central Blood Ctr., Tokyo, Japan. 2. Dept. of Human Genetics, Univ. of Tokyo, Tokyo, Japan. 3. Transplantation Immunology Unit, Centre Medical Univ., Geneva, Switzerland. 4. Harbin Red Cross Blood Ctr., Harbin, China. 5. Nt!. Ctr. of Anthlopology, Ulaanbaatar, Mongolia. We established a PCR-SSOP method which can be used to distinguish all HLA-A2 alleles (A*0201-A*0217). After serological typing for A2, genomic DNA was PCR amplified with A2-specificprimers, and the amplified fragment was hybridized with 24 kinds of oligonucleotide probes designed on exons 2, 3 and 4. Using this method, A2 allelic subtypes were determined in Japanese (n=114), Chinese (Korean (n=100), Manchurian (n=100» and Mongolian (n=IOO) population samples. A total of five alleles, A*0201, A*0203, A*0205, A*0206 andA*0207, were detected in this study. ill all populations,A*0201 and A*0206 were the most frequent A2 subtypes and accounted for 50-60% and 2035%, respectively, of the A2 alleles. These alleles did not exhibit any strong associationswith other loci. A*OZ07 was observedwith relativelyhigh frequency in Japanese, Korean and Manchurianand was associatedwith Cw'0102-B*4601DRBI'0803 ouly in the Japanese (4n). ill Mongolian,the frequency of A*OZ05 was higher than that of A*0207, and A*0205 was stronglyassociatedwith Cw6B50-DR7 (518). ill Manchurian, A'OZ03 occured with relatively high frequency and the allele was associatedwith B52 and DRBI*1404 (5/6).
0262 sSP/ARMS ANALYSIS OF HLA CLASS I ALLELES OF SAMOANS
NielsGrunnet'P, RudiSteffensen', Kim varmlng', and CasperJersild'.
, Regional Center for Blood Transfusion andClinical Immunology, Aalborg Hospital, Aalborg, Denmark. 2 Present address: Tissue typing laboratory. Department ofClinical Immunology, Aarhus University Hospital, Skejby, Denmark. Theaimof Ibisstudy was to point out the possible ethnic differences of HLA class I genotypes between a group of 43 heallby unrelated Inuits, born in Greenland with residence in Denmark and a gronp nf 30 healthy unrelated Danes. We studied the genotype frequencies of HLA-A-B andC by polymerase chain reaction with sequence specific primers in a amplification refractory mutation system (SSP ARMS-PeR)
suppLemented with nested PCR reagents using XIIworkshop reagents 10 determine the frequencies of HLA-A*02subtypes. Significant differences (p <0_05) wereobserved for several antigens. Concerning the A locus, the frequency of A*Z4(9) was significantly higher inInuits Iban inDanes (0.57/0.13. p<0.OOO5) while A*OI wassignificantly lower inInuits Iban in Danes (0.047/0.167. p=0.033).TheA*OZ antigen occurred with almost equal frequency (0.Z44/0.267 p=0.905)butthefrequency of thesubtype A*OZOI were significant less frequent andA*OZ06 significant higher in Inuits (0.116/0.267, p~O.035 and 0.128/0, p=0.0045 respectively), ForIbeBlocus, B*44(12) occurred significantly lessfrequent andB*48 signiflcantLy more frequent in Inuits (0.047/0.18, p=0.017and 0.116/0.0. p=0.OO8. respectively). Finally, four differences were observed in the C loCus, C*03 and C·08 were significantly more frequent in Inuits compared to Danes (0.384/0,167, p=0.OO70 and O. I16/0,p~O,OO80, respectively) andC*06 andC*07 were
significant less frequent in Inuits (0.0/0.083, p=O.OZl and lZ8/0.383. p= 0.0007. respectively). Theresults ofIbisstudy confirm previous published data based onserology from XI Workshop. It also provides further evidence of more extensive differences in
HLA class I polymorphism between these twopopulations.
Laura D. Severson, Douglas E. Crews, and Raymond W. Lang The Ohio State University, Columbus, Ohio USA Samoa is a group of volcanic islands lying in the South Pacific about 150 below the equator and just east of the international date line. Mainly Polynesian, Samoans are believed to be descended from Austronesianspeaking populations which entered Melanesia from eastern Indonesia approximately 3,500 years ago and migrated east to Samoa. HLA Class I alleles were typed using sequence-specific primer (SSP) combinations in a one-step PCR system based on the amplification refractory mutation system (ARMS). Identification of alleles is based on presence or absence of amplified product after agarose gel electrophoresis. This methodology is being used to assess the Class I allelic distribution in a group (280) samples of American Samoans. Initial results show a high frequency of A02, AIIOI, A2402, A26, A3401; B3801, B3901, B40, B48, 85501; CwOIOI, Cw03, CW0401,CW0701,CW0801,and Cw150l . which is consistent with previous reports from Oceania using serologic techniques. This first report on HLAC allelic distribution of American Samoans shows results similarto those for other Polynesian populations. In addition, this work has identified allelic subtypes not revealed by serology in Polynesians, thus providing finer detail for assessing ancestral affinitiesamong these populations.
Abstracts
11.1. Anthropology Today: Contribution of HLA (I) 0258
0257
DNA ITPING OF HLA CLASS [ GENES REVEALS FURTHER GENETIC DIFFERENCE BE1WEEN NORl'HERN AND SOUTHERN CHINESE
HIGH RESOLUTION MOLECULAR TYPING OF HLA-A AND HLA-B IN THREE SOUTH AMERICAN INDIAN TRIBES
Xiaofang Sun, YipingSun. Xiaojiang Gao Guangzhou Second People's Hospital, Guangzhou, China, The Australian National University, Canberra, Australia
Fernandez-Vitia MA, Lazaro AM, Marcos YC, NulfCJ, Fish VM, McGarry JE, Raimondi EH, Stastny P, Dept oflnternal Medicine, UT Southwestern Medical Ctr, Dallas, TX, USA
Our previous serological and class II DNAanalyses have demonstrated markedly different HLA profiles in the northern and southern Chinese.
We analyzed 187 unrelated individuals from the Toba (TOB, n=122), Wichi (WlC, n=47) and Pilaga (pIL n~18) tribes from NE Argentina. The number of alleles at the HLA-A locus was small (TOB n= 6, WlC n=6 and PIL n=4). In contrast, the HLA-B locus presented larger allelic variation (TOB n=23, WlC n=13, PIL n~14). HLA-A alleles consisted of A*0201, 2402,2403,68012,3101. A putative new allele, locally called A-02X09, with a hybridization pattern similar to A*020 I, but differing in codons 142167 in which it is similarto A*2402, was observed in TOB and WlC. All HLA-A locus alleles had similar gene frequencies which resulted in low homozygosity frequencies (hgfTOB= 0.18, WlC= 0.20, PIL=0.25). In TOB we observed two subtypes ofHLA- B5, three ofB35, 6 ofBl5, three ofB48, three ofB39 and 5 ofB40. We observed several new hybridization patterns which appeared to correspond to new alleles. Among them was a new subtype ofB48 which was the most frequent B-locus allele in this population (gf=0.15). In these inbred populations, in which polymorphism ofHLA-A and 8 loci is restricted, balanced allelic frequencies at both Class I loci appear to have resulted from selection for heterozygosity. However, a larger polymorphism and more frequent generation of new aUeles at the HLA-B locus. suggests that different evolutionary forces may have acted on each of these HLA Class I loci.
In the present experiments using comprehensive DNA typing protocols
developed in our laboratory we examined the HLA-A and -B genes in 85 northern (Beijing) and 160 southern [Guangdong) Chinese donors for serologically identical subtypes. Several antigens showed significantly different distrtbutions including Al, A26, A30, A3!. B37. B44. B48, B52 and B61 predominantly found in the north and All. B38. B46, B55, and B58 predominantly or only detected in the south. A2 the only A antigen further split by DNA typing showed extraordinary heterogeneity with six subtypes detected including A"0201, 0203, 0205, 0206. 0207 and 0210. In the B locus, however, 1/3 of the serological specificities were found to have hidden subtypes. Further, several antigens commonly seen in
both groups were found to be different DNAsubtypes including A"0201, B'0702, 1302, 1501, 2705, and 4006 in the north and A'0203, 0207, 8'0705, 130!. 1502, 2704 and 4001 in the south. In accordance with our previous reports these results reflect historical population mixtures with different genetic inputs in northern and southern China. The
previously
undetectable
hidden
subtypes
may have
important
implications for unrelated-donor transplantations in these people.
0259 HLA
CLASS
I
AND
CLASS
II
ALLELES
IN
THREE
SYMPATRIC WEST AFRICAN ETHNIC GROUPS Gaia LuonP·2, Maria De Luca1 ,3, David Modiano-, Mario Coluzzl'', Steven
GEMarsh! & JuliaG Boomer"'Tissue Antigen Laboratory, Imperial Cancer Research Fund,London, UK 21nstitute of Parasilology, University "LaSapienza", Roma, Italy 'Dept. ofCellular Biology. University ofCalabria, Arcavacala di Rende, Ilaly We havecollected a totalof 463samples from three West African populations: 187 Massi, 160 Rimaibe and 116 Fulani. These three elhnicgroupsare sympatric and have totally diflerent genelicand cultural backgrounds; while Mossi and Birnaioe are a Sudanese Negroid sedentary population, the Fulaniare descended from nomadic pasioralists and have non-Negroid features of possibleCaucasianorigin. Inorder to investigate the past and present genetic relationships betweenthese Ihree populations we are typing these samples [or HLA Class I using the 12th Workshop SSP ARMS-PCR typing kit and for HLA Class II using the 12th Workshop SSG-PCRtechnique. Preliminary data indicates a highfrequency of Cw·1701. Cw'04. B'5301 and B'4201 alleles in the Mossi tribe. Data will be presented on the allele and haplotype frequencies of all three populations, together with comparisons of genetic distance to other African and European populations,
0261
HLA class I polymorphism in Inuits.
0260 HLA-A2ALLELES IN NORTHEAST-ASIAN POPULATIONS Yoshihide Ishikawa', Katsushi Tokunaga'r, Jean-Marie Tiercy', Kouichi Kashiwase', Hidenori Tanaka', Jie Liu', Tatsuya Akaza', Kenji Tadokoro', Nyarn-Osorin Chimge",Guan JIUI Jia', Takeo Juji' I. Japanese Red Cross Central Blood Ctr., Tokyo, Japan. 2. Dept. of Human Genetics, Univ. of Tokyo, Tokyo, Japan. 3. Transplantation Immunology Unit, Centre Medical Univ., Geneva, Switzerland. 4. Harbin Red Cross Blood Ctr., Harbin, China. 5. Nt!. Ctr. of Anthlopology, Ulaanbaatar, Mongolia. We established a PCR-SSOP method which can be used to distinguish all HLA-A2 alleles (A*0201-A*0217). After serological typing for A2, genomic DNA was PCR amplified with A2-specificprimers, and the amplified fragment was hybridized with 24 kinds of oligonucleotide probes designed on exons 2, 3 and 4. Using this method, A2 allelic subtypes were determined in Japanese (n=114), Chinese (Korean (n=100), Manchurian (n=100» and Mongolian (n=IOO) population samples. A total of five alleles, A*0201, A*0203, A*0205, A*0206 andA*0207, were detected in this study. ill all populations,A*0201 and A*0206 were the most frequent A2 subtypes and accounted for 50-60% and 2035%, respectively, of the A2 alleles. These alleles did not exhibit any strong associationswith other loci. A*OZ07 was observedwith relativelyhigh frequency in Japanese, Korean and Manchurianand was associatedwith Cw'0102-B*4601DRBI'0803 ouly in the Japanese (4n). ill Mongolian,the frequency of A*OZ05 was higher than that of A*0207, and A*0205 was stronglyassociatedwith Cw6B50-DR7 (518). ill Manchurian, A'OZ03 occured with relatively high frequency and the allele was associatedwith B52 and DRBI*1404 (5/6).
0262 sSP/ARMS ANALYSIS OF HLA CLASS I ALLELES OF SAMOANS
NielsGrunnet'P, RudiSteffensen', Kim varmlng', and CasperJersild'.
, Regional Center for Blood Transfusion andClinical Immunology, Aalborg Hospital, Aalborg, Denmark. 2 Present address: Tissue typing laboratory. Department ofClinical Immunology, Aarhus University Hospital, Skejby, Denmark. Theaimof Ibisstudy was to point out the possible ethnic differences of HLA class I genotypes between a group of 43 heallby unrelated Inuits, born in Greenland with residence in Denmark and a gronp nf 30 healthy unrelated Danes. We studied the genotype frequencies of HLA-A-B andC by polymerase chain reaction with sequence specific primers in a amplification refractory mutation system (SSP ARMS-PeR)
suppLemented with nested PCR reagents using XIIworkshop reagents 10 determine the frequencies of HLA-A*02subtypes. Significant differences (p <0_05) wereobserved for several antigens. Concerning the A locus, the frequency of A*Z4(9) was significantly higher inInuits Iban inDanes (0.57/0.13. p<0.OOO5) while A*OI wassignificantly lower inInuits Iban in Danes (0.047/0.167. p=0.033).TheA*OZ antigen occurred with almost equal frequency (0.Z44/0.267 p=0.905)butthefrequency of thesubtype A*OZOI were significant less frequent andA*OZ06 significant higher in Inuits (0.116/0.267, p~O.035 and 0.128/0, p=0.0045 respectively), ForIbeBlocus, B*44(12) occurred significantly lessfrequent andB*48 signiflcantLy more frequent in Inuits (0.047/0.18, p=0.017and 0.116/0.0. p=0.OO8. respectively). Finally, four differences were observed in the C loCus, C*03 and C·08 were significantly more frequent in Inuits compared to Danes (0.384/0,167, p=0.OO70 and O. I16/0,p~O,OO80, respectively) andC*06 andC*07 were
significant less frequent in Inuits (0.0/0.083, p=O.OZl and lZ8/0.383. p= 0.0007. respectively). Theresults ofIbisstudy confirm previous published data based onserology from XI Workshop. It also provides further evidence of more extensive differences in
HLA class I polymorphism between these twopopulations.
Laura D. Severson, Douglas E. Crews, and Raymond W. Lang The Ohio State University, Columbus, Ohio USA Samoa is a group of volcanic islands lying in the South Pacific about 150 below the equator and just east of the international date line. Mainly Polynesian, Samoans are believed to be descended from Austronesianspeaking populations which entered Melanesia from eastern Indonesia approximately 3,500 years ago and migrated east to Samoa. HLA Class I alleles were typed using sequence-specific primer (SSP) combinations in a one-step PCR system based on the amplification refractory mutation system (ARMS). Identification of alleles is based on presence or absence of amplified product after agarose gel electrophoresis. This methodology is being used to assess the Class I allelic distribution in a group (280) samples of American Samoans. Initial results show a high frequency of A02, AIIOI, A2402, A26, A3401; B3801, B3901, B40, B48, 85501; CwOIOI, Cw03, CW0401,CW0701,CW0801,and Cw150l . which is consistent with previous reports from Oceania using serologic techniques. This first report on HLAC allelic distribution of American Samoans shows results similarto those for other Polynesian populations. In addition, this work has identified allelic subtypes not revealed by serology in Polynesians, thus providing finer detail for assessing ancestral affinitiesamong these populations.