Feasibility of using genetic linkage analysis to identify the genes encoding T cell-defined minor histocompatibility antigens.
Feasibility of using genetic linkage analysis to identify the genes encoding T cell-defined minor histocompatibility antigens.
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使用遗传连锁分析来鉴定编码 T 细胞定义的次要组织相容性抗原的基因的可行性。
DOI：
10.1034/j.1399-0039.2002.590407.x
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Riddell,SR
中科院分区：
文献类型：
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作者：
Warren,EH;Otterud,BE;Linterman,RW;Brickner,AG;Engelhard,VH;Leppert,MF;Martin,PJ;Riddell,SR
我们已经评估了遗传连锁分析的效用，以确定编码次要组织相容性抗原的基因，使用牛痘病毒载体作为一种简单方便的方法，在淋巴母细胞系中瞬时表达I类MHC分子。作为一个测试案例，我们使用了一个识别HA-8的CTL克隆，HA-8是一种由KIAA 0020基因编码并由HLA-A *0201呈递的次要组织相容性抗原。用编码HLA-A *0201转基因的重组牛痘病毒载体感染来自CEPH参考家族收集的三个大谱系中的个体的EBV转化的B细胞系，这导致MHC限制性等位基因HLA-A *0201在细胞表面上的高水平表达。然后使用标准体外细胞毒性测定法测定牛痘感染的靶细胞中的HA-8表达。对这些家系中HA-8表达分离的成对连锁分析表明，HA-8基因与位于染色体9 p远端部分的一簇标记位点紧密连锁。对三个家族中个体的9 p标记单倍型的分析鉴定了具有重组单倍型的几个个体，并且这些重组事件用于进一步改进HA-8基因定位的精确度。这些数据共同表明，HA-8基因定位于远端9 p的10.3cM（对应于3.9 Mb）间隔，被认为编码至少11个基因，包括KIAA 0020。 这些结果表明，连锁分析可以用于定位次要组织相容性基因，具有较高的精度和准确性。在接下来的几年里，人类基因组序列的细化和注释无疑将增加连锁分析作为识别次要组织相容性抗原基因的工具的实用性。
We have evaluated the utility of genetic linkage analysis to identify genes that encode minor histocompatibility antigens using vaccinia virus vectors as a simple and convenient method for transient expression of class I MHC molecules in lymphoblastoid cell lines. As a test case, we used a CTL clone that recognizes HA‐8, a minor histocompatibility antigen encoded by the KIAA0020 gene and presented by HLA‐A*0201. EBV‐transformed B cell lines from individuals in three large pedigrees from the CEPH reference family collection were infected with a recombinant vaccinia virus vector encoding an HLA‐A*0201 transgene, which led to high level expression of the MHC restricting allele HLA‐A*0201 on the cell surface. HA‐8 expression in the vaccinia‐infected target cells was then determined using standardin vitrocytotoxicity assays. Pairwise linkage analysis of the segregation of HA‐8 expression in these pedigrees demonstrated that the HA‐8 gene was tightly linked with a cluster of marker loci located on the distal portion of chromosome 9p. Analysis of 9p marker haplotypes for individuals in the three families identified several individuals with recombinant haplotypes, and these recombination events were used to refine the precision of the HA‐8 gene localization further. The data collectively indicate that the HA‐8 gene is localized to a 10.3 cM (corresponding to 3.9 Mb) interval of distal 9p that is thought to encode at least 11 genes, including KIAA0020. These results demonstrate that linkage analysis can be used to map minor histocompatibility genes with high precision and accuracy. Over the next years, refinement and annotation of the human genome sequence will undoubtedly increase the utility of linkage analysis as a tool for identifying minor histocompatibility antigen genes.