四川骨髓库汉族人群HLA-DPB1等位基因的分布及HLA-DPB1在HSCT中错配模式的初步研究

发布时间：2018-02-26 04:24

本文关键词： HLA-DPB1 四川汉族测序高分辨分型禁忌错配　出处：《北京协和医学院》2015年硕士论文　论文类型：学位论文

【摘要】：[目的]调查四川骨髓库汉族人群HLA-DPB1等位基因多态性分布情况,估计四川骨髓库汉族人群非血缘造血干细胞移植中可能存在的HLA-DPB1错配情况及错配模式,同时分析HLA-A,-B,-DRB1全相合的同胞间HLA-DPB1位点存在的错配情况及错配模式。[方法]从四川骨髓库随机抽取1097份四川汉族非血缘造血干细胞移植捐献者DNA样本,从四川省脐带血造血干细胞库和重庆市血液中心临床移植配型供受者家系中筛选同胞间HLA-A,-B,-DRB1全相合的移植家系901个。采用PCR-SBT技术对DNA样本进行HLA-DPB1测序分型。经ABI 3730测序仪测序,用UTYPE6.0软件分析结果。对于因第二外显子区域内的Trans/cis连锁产生的模棱两可结果,设计组特异性测序引物(Group Specific Sequencing Primers, GSSPs)进行单等位基因测序。对于区别在第二外显子外的模棱两可结果,以G为后缀表G组,指定分型结果。用直接计数法统计四川骨髓库汉族人群中HLA-DPB1等位基因的基因频率；用SPSS19.0软件对不同人群HLA-DPB1等位基因分布进行X2检验。采用T细胞表位(T-cell-epitope, TCE) 3/4模型理论推测四川汉族非血缘造血干细胞移植供受者之间及HLA-A,-B,-DRB1全相合的同胞之间可能存在的HLA-DPB1等位基因错配模式。[结果](1)在四川骨髓库汉族人群中共检出35种HLA-DPB1等位基因型别,较常见的等位基因及其频率分别是：HLA-DPB1*05:01:01G(gene frequency, GF=40.9%)、*02:01:02G(GF=16.1%)、*02:02(GF=8.4%)、* 13:01:01G(GF=7.9%)、*04：01：01G(GF=6.8%)等,这5种等位基因共占80.1%。35种等位基因中检出9种美国组织相容性和免疫遗传学协会(American Society for Histocompatibility and Immunogenetics, ASHI)常见及确认等位基因(Common and Well-Documented, CWD)表中尚不包含的等位基因,分别是：DPB1*41：01：01(5例),DPB1*48：01(3例),DPB1*100：01(3例),DPB1*04:01:15、DPB1*25:01、DPB1*71:01、DPB1*91:01、DPB1*93：01和DPB1*331：01各1例。还有2例新等位基因,世界卫生组织命名委员会将其分别命名为DPB1*363：01和DPB1*394：01。四川骨髓库汉族人群与广州汉族人群HLA-DPB1等位基因分布之间的差异不具有统计学意义(X2=21.556,P0.01),而与山东汉族(X2=51.134,P0.01)及高加索人群(X2=712.92,P0.01)之间的差异均具有统计学意义。(2)按照TCE模型对四川骨髓库汉族人群HLA-DPB1等位基因进行分组,高免疫原性组占2.19%,中等免疫原性组占8.89%,低免疫原性组占88.92%。四川汉族人群中进行非血缘造血干细胞移植时,根据TCE模型推测HLA-DPB1可能存在的允许错配占34.28%,在GvH方向和HvG方向的禁忌错配分别为17.27%,TCE3模型允许而TCE4模型不允许的错配在GJvH方向和HvG方向分别为15.59%。(3)移植家系研究中发现DPB1位点交换率为5.2%。允许错配占交换家系53%；另外47%为禁忌错配。]HLA-DPB1禁忌错配在HLA-A,-B,-DRB1全相合的同胞中占2.44%。[结论]四川骨髓库汉族人群中HLA-DPB1高频等位基因分布集中,多态性较强,并具有自身的分布特点。根据TCE模型分析表明,在四川汉族非血缘造血干细胞移植时可能存在HLA-DPB1的禁忌错配,提示临床需要对此重视。根据TCE模型分析发现,在HLA-A,-B,-DRB1全相合的同胞间可能存在HLA-DPB1的交换,在进行同胞间造血干细胞移植时,也可能存在HLA-DPB1的禁忌错配。
[Abstract]:Objective: To investigate the Sichuan bone marrow bank Han population HLA-DPB1 gene polymorphism distribution of Sichuan Han population estimate bone marrow unrelated hematopoietic stem and mismatch HLA-DPB1 mismatch patterns may exist in the cell transplantation, and the analysis of the HLA-A, -B, -DRB1 identical sibling mismatch and HLA-DPB1 loci are wrong distribution mode. Methods from Sichuan marrow library on a random sample of 1097 Sichuan Han unrelated hematopoietic stem cell donor DNA samples from Sichuan Province, umbilical cord blood stem cell bank and Chongqing Blood Center for clinical transplantation typing by HLA-A screening, sibling incidencein family -B, -DRB1 matched transplantation in 901 families a sample of DNA. HLA-DPB1 genotyping using PCR-SBT technology. The ABI 3730 sequencing, the results were analyzed with UTYPE6.0 software. As a result of the second exon region of the Trans/cis chain from the two edge Can the design of group specific primers (Group Specific Sequencing Primers, GSSPs) by single allele sequencing. Results for the difference in the ready to accept either course of exon 2 of G group with the G suffix table, specify the typing results. The gene frequency of HLA-DPB1 by direct counting statistics of Sichuan marrow Library in Han population gene; X2 test was performed on different populations of HLA-DPB1 alleles with SPSS19.0 software. The T cell epitopes (T-cell-epitope, TCE) 3/4 model theory between Sichuan Han unrelated hematopoietic stem cell transplantation by HLA-A and -B, may exist between -DRB1 identical sibling HLA-DPB1 allele. With the model. Results (1) in Sichuan Han population were detected in the bone marrow of 35 type of HLA-DPB1 allele, common alleles and their frequencies are: HLA-DPB1*05:01:01G (gene frequency, GF= 40.9%), *02:01:02G (GF=16.1%), *02:02 (GF=8.4%), * 13:01:01G (GF=7.9%), *04:01:01G (GF=6.8%), a total of 5 alleles of 9 American histocompatibility and immunogenetics association detection 80.1%.35 allele (American Society for Histocompatibility and Immunogenetics, ASHI) and common confirmation a gene (Common and Well-Documented, CWD) in the table is not contained in the alleles were DPB1*41:01:01 (5 cases), DPB1*48:01 (3 cases), DPB1*100:01 (3 cases), DPB1*04:01:15, DPB1, *25:01, DPB1*71:01, DPB1*91:01, DPB1*93:01 and DPB1*331:01 of the 1 cases. There were 2 new alleles. The committee will be named WHO named as the difference between the distribution of DPB1*363:01 and DPB1*394:01. in Sichuan Han population of bone marrow and HLA-DPB1 in Guangzhou Han population allele was not statistically significant (X2=21.556 P0.01, and Shandong), Han (X2=51.134, P0.01) and Caucasian (X2=712.92, P0.01) between the differences were statistically significant. (2) according to the TCE model of Sichuan marrow library Han population allele HLA-DPB1 group, high immunogenicity group accounted for 2.19%, in the immunogenicity of the group accounted for 8.89%, low immunogenicity was unrelated hematopoietic stem cell transplantation for 88.92%. in Sichuan Han population, according to the TCE model suggests that HLA-DPB1 may exist to allow mismatches accounted for 34.28%, the taboo mismatch in GvH and HvG directions respectively for 17.27% TCE3, while the TCE4 model allows the model does not allow the mismatch in the GJvH direction and the direction of HvG were 15.59%. (3) DPB1 transplantation sites exchange rate is allowed 5.2%. mismatch 53% families accounted for exchange found in family studies; the other 47% with.]HLA-DPB1 mismatch in the wrong taboo taboo HLA-A, -B, -DRB1 matched with cells accounted for 2.44%.[Conclusion] four HLA-DPB1 high frequency allele distribution in bone marrow in Han population, strong polymorphism, and has its own characteristics. According to the distribution of the TCE model showed that the taboo mismatch HLA-DPB1 may exist in Sichuan Han unrelated hematopoietic stem cell transplantation, tips for this attention to clinical needs. According to the TCE model analysis found that in HLA-A -B, there may be HLA-DPB1 exchange -DRB1 identical sibling, the sibling hematopoietic stem cell transplantation, taboo mismatches may also exist in HLA-DPB1.

【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R457.7

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