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106-P: The Analysis of MICA Allele and Transmembrane Region Polymorphism in Chinese Han Population
Abstracts
S85
105-P
THE MACAQUE MHC-DQB1 LOCUS IS A POLYMORPHIC SINGLE COPY GENE WITH ALLELE GROUPS THAT ARE CLOSELY RELATED TO THEIR HLA-DQB1 ALLELE GROUP COUNTERPARTS. Jin Wu, Sue Bassinger, Tenzin D. Tsewang, Michael A. Krencicki, Jr., Jennifer B. Woods, Thomas M. Williams. Pathology, University of New Mexico, Albuquerque, NM, USA. Aim: Non-human primates are important as models for infectious disease, vaccine development, and transplantation studies. The macaque MHC DQB1 locus is not very well understood. Our goal is to develop high-throughput, high resolution, and robust genotyping molecular tools to characterize rhesus, pig-tail, and cynomolgus macaque DQB1 genes. Methods: Two sets of group specific PCR primers were designed and linked to M13 universal primers to amplify either Mamu-DQB1*06 or Mamu-DQB1*15/16/17/18/24 groups from rhesus genomic DNA. Exon 2 was directly sequenced with M13 forward and reverse primers. These primers were also used to amplify and sequence DQB1 alleles from pig-tail and cynomolgus genomic DNA. Sequences were analyzed with Assign software (Conexio, Perth, AU). Results: We used this method to genotype about 300 rhesus, pig-tail, and cynomolgus macaques. Fourteen novel and 32 known alleles were identified in rhesus, 31 novel alleles in pig-tail, and 26 novel and 22 known alleles in cynomolgus macaques. Some allele sequences are identical across all 3 species. We were able to define DQB1 allele frequencies for these animals. Non-human primate MHC studies have shown that Class I A and B genes are duplicated loci with multiple A and B genes. We have not observed that the primate DQB1 gene is duplicated. In addition, our phylogenetic tree analysis shows that the primate DQB1 allele groups are closely related to their HLA DQB1 allele group counterparts. Conclusions: Our data show that the rhesus, pig-tail, and cynomolgus macaque DQB1 locus is highly polymorphic and shares many similarities with HLA DQB1.
106-P
THE ANALYSIS OF MICA ALLELE AND TRANSMEMBRANE REGION POLYMORPHISM IN CHINESE HAN POPULATION. Xiaoni Yuan, Jun He, Xiaojing Bao, Chao Xu. Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China. Aim: The MICA gene polymorphism plays an important role in allogenic organ transplantation and hematopoietic stem cell transplantation. We analyzed to further assess of MICA polymorphism in the 137 Chinese Han healthy people. Methods: All samples were analyzed using PCR-SSOP to measure the MICA gene exons 2, 3, 4, 5. The ambiguous alleles combination were distinguished by SBT. Meanwhile the MICA gene transmembrane region microsatellite sequence were detected by the SBT specific primer. Results: From 137 Chinese Han healthy people, 13(19.40%) MICA alleles were detected in the 61 various MICA alleles. MICA*008(23.36%) was the most common alleles, and then were MICA*010(21.53%), MICA*002 (19.34%), MICA*009(10.58%), MICA*027(7.30%), MICA*012(6.93%), MICA*016 (5.11%), MICA*007 (2.55%), MICA*045 (2.19%), MICA*004 (1.46%), MICA*017(1.46%), respectively. 42 allele combination were detected, and most common alleles of MICA*008, MICA*010, MICA*002, MICA*009 and MICA*027 respectively combination accounted for 63.51%. The common allele combination were MICA*008, MICA*010 (10.95%); MICA*008,MICA*008(9.49%); MICA*008, MICA*027 (5.11%); MICA*010, MICA*010 (9.49%); MICA*002, MICA009(5.84%). In the MICA transmembrane region types A5, A5.1, A9, A6, A4 were found, the frequency were A5(33.94%), A5.1(23.36%), A9(20.80%), A6(12.41%), A4(12.04%). The combination between A5, A5.1, A9 constituted 49.64% of transmembrane region types. A5 associated with A5.1 or A9 were very common, account for 16.79%, 13.14% and 10.22%. Conclusions: The combination of PCR-SSOP and SBT can provide an accurate and reliable method to detect allele distribution of MICA polymorphism. The distribution of MICA polymorphism in Chinese population are relatively focused. Our results provide frequency distribution data about MICA allele polymorphism in Chinese Han population, The characteristic of MICA gene polymorphism is helpful to our future research. Yuan: The First Affiliated Hospital of Soochow University: Science Med Advisor. He: The First Affiliated Hospital of Soochow University: Science Med Advisor. Bao: The First Affiliated Hospital of Soochow University: Science Med Advisor. Xu: The First Affiliated Hospital of Soochow University: Science Med Advisor.
S85
105-P
THE MACAQUE MHC-DQB1 LOCUS IS A POLYMORPHIC SINGLE COPY GENE WITH ALLELE GROUPS THAT ARE CLOSELY RELATED TO THEIR HLA-DQB1 ALLELE GROUP COUNTERPARTS. Jin Wu, Sue Bassinger, Tenzin D. Tsewang, Michael A. Krencicki, Jr., Jennifer B. Woods, Thomas M. Williams. Pathology, University of New Mexico, Albuquerque, NM, USA. Aim: Non-human primates are important as models for infectious disease, vaccine development, and transplantation studies. The macaque MHC DQB1 locus is not very well understood. Our goal is to develop high-throughput, high resolution, and robust genotyping molecular tools to characterize rhesus, pig-tail, and cynomolgus macaque DQB1 genes. Methods: Two sets of group specific PCR primers were designed and linked to M13 universal primers to amplify either Mamu-DQB1*06 or Mamu-DQB1*15/16/17/18/24 groups from rhesus genomic DNA. Exon 2 was directly sequenced with M13 forward and reverse primers. These primers were also used to amplify and sequence DQB1 alleles from pig-tail and cynomolgus genomic DNA. Sequences were analyzed with Assign software (Conexio, Perth, AU). Results: We used this method to genotype about 300 rhesus, pig-tail, and cynomolgus macaques. Fourteen novel and 32 known alleles were identified in rhesus, 31 novel alleles in pig-tail, and 26 novel and 22 known alleles in cynomolgus macaques. Some allele sequences are identical across all 3 species. We were able to define DQB1 allele frequencies for these animals. Non-human primate MHC studies have shown that Class I A and B genes are duplicated loci with multiple A and B genes. We have not observed that the primate DQB1 gene is duplicated. In addition, our phylogenetic tree analysis shows that the primate DQB1 allele groups are closely related to their HLA DQB1 allele group counterparts. Conclusions: Our data show that the rhesus, pig-tail, and cynomolgus macaque DQB1 locus is highly polymorphic and shares many similarities with HLA DQB1.
106-P
THE ANALYSIS OF MICA ALLELE AND TRANSMEMBRANE REGION POLYMORPHISM IN CHINESE HAN POPULATION. Xiaoni Yuan, Jun He, Xiaojing Bao, Chao Xu. Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China; Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China. Aim: The MICA gene polymorphism plays an important role in allogenic organ transplantation and hematopoietic stem cell transplantation. We analyzed to further assess of MICA polymorphism in the 137 Chinese Han healthy people. Methods: All samples were analyzed using PCR-SSOP to measure the MICA gene exons 2, 3, 4, 5. The ambiguous alleles combination were distinguished by SBT. Meanwhile the MICA gene transmembrane region microsatellite sequence were detected by the SBT specific primer. Results: From 137 Chinese Han healthy people, 13(19.40%) MICA alleles were detected in the 61 various MICA alleles. MICA*008(23.36%) was the most common alleles, and then were MICA*010(21.53%), MICA*002 (19.34%), MICA*009(10.58%), MICA*027(7.30%), MICA*012(6.93%), MICA*016 (5.11%), MICA*007 (2.55%), MICA*045 (2.19%), MICA*004 (1.46%), MICA*017(1.46%), respectively. 42 allele combination were detected, and most common alleles of MICA*008, MICA*010, MICA*002, MICA*009 and MICA*027 respectively combination accounted for 63.51%. The common allele combination were MICA*008, MICA*010 (10.95%); MICA*008,MICA*008(9.49%); MICA*008, MICA*027 (5.11%); MICA*010, MICA*010 (9.49%); MICA*002, MICA009(5.84%). In the MICA transmembrane region types A5, A5.1, A9, A6, A4 were found, the frequency were A5(33.94%), A5.1(23.36%), A9(20.80%), A6(12.41%), A4(12.04%). The combination between A5, A5.1, A9 constituted 49.64% of transmembrane region types. A5 associated with A5.1 or A9 were very common, account for 16.79%, 13.14% and 10.22%. Conclusions: The combination of PCR-SSOP and SBT can provide an accurate and reliable method to detect allele distribution of MICA polymorphism. The distribution of MICA polymorphism in Chinese population are relatively focused. Our results provide frequency distribution data about MICA allele polymorphism in Chinese Han population, The characteristic of MICA gene polymorphism is helpful to our future research. Yuan: The First Affiliated Hospital of Soochow University: Science Med Advisor. He: The First Affiliated Hospital of Soochow University: Science Med Advisor. Bao: The First Affiliated Hospital of Soochow University: Science Med Advisor. Xu: The First Affiliated Hospital of Soochow University: Science Med Advisor.