ANTIGENIC STRUCTURE OF HLA-B27

几种常见的人类疾病状态，包括强直性 脊柱炎、感染后反应性关节炎和急性前路 葡萄膜炎与血清学标志物--人类白细胞抗原- B27.令人信服的证据已经积累起来，一班的专业 细胞上鉴定为人类白细胞抗原B27的组织相容性(MHC)抗原 表面蛋白直接参与了疾病的发病机制。这个 这种参与的机制尚不清楚，但可能与 与人类白细胞抗原B27分子独特的抗原结构有关。这个 拟议实验的总体目标是扩大 基本上现有的关于抗原性的知识 人类白细胞抗原B27分子的结构。 这将通过四个具体目标来实现： 1)将产生一组突变的B27基因 寡核苷酸介导的定点突变；这些突变体 基因将通过氨基酸替换来编码B27分子 预测在同种异体抗原决定簇中起重要作用的位点 是B27的。然后，这些分子将在体外细胞中表达。 2)表达野生型HLA-B27基因的转基因小鼠， 突变的HLAB27基因或相关的人类白细胞抗原B7基因将被创造出来 通过显微注射受精卵。3)单抗 人类白细胞抗原B27的限制性特异性将由转基因产生 表达B7基因或突变B27基因的小鼠免疫 来自野生型B27转基因小鼠的细胞。4)这些 抗体，以及其他单抗和多克隆抗B27 抗体，将被测试与B27突变分子的结合， 由此产生的反应模式将被用来产生 用计算机辅助绘制天然B27的抗原性洗涤剂图谱 分子，基于最近发表的结晶学图谱 人类白细胞抗原-A2。预计该项目将提供 充足的资源和信息，用于合理设计 B27相关致病机制的成功实验模型 疾病。

Several common human disease states, including ankylosing spondylitis, post-infectious reactive arthritis, and acute anterior uveitis, are genetically associated with the serologic marker HLA- B27. Persuasive evidence has accumulated that the class 1 major histocompatibility (MHC) antigen identified as HLA-B27 on cell surfaces is directly involved in disease pathogenesis. The mechanism of this involvement is unknown, but is presumably related to the unique antigenic structure of the HLA-B27 molecule. The overall goal of the proposed experiments is to enlarge substantially the existing knowledge regarding the antigenic structure of the HLA-B27 molecule. This will be accomplished through four specific aims: 1) A panel of mutant B27 genes will be produced by oligonucleotide-mediated site-directed mutagenesis; these mutant genes will encode B27 molecules with amino acid substitutions at sites predicted to be important in the alloantigenic determinants of B27. These molecules will then be expressed in cells in vitro. 2) Transgenic mice expressing either the wild-type HLA-B27 gene, the mutant HLAB27 genes, or the related HLA-B7 gene will be created by microinjection of fertilized ova. 3) Monoclonal antibodies with restricted specificity for HLA-B27 will be generated from trangenic mice expressing either the B7 gene or mutant B27 genes, immunized with cells from the wild-type B27 transgenic mice. 4) These antibodies, along with other monoclonal and polyclonal anti-B27 antibodies, will be tested for binding to the B27 mutant molecules, and the resulting patterns of reactivity will be used to generate a computer-assisted map of antigenic determirants on the native B27 molecule, based on the recently published crystallographic map of HLA-A2. It is anticipated that this project will provide sufficient resourses and information for the rational design of successful experimental models of the pathogenesis of B27-related disease.