ANTIGENIC STRUCTURE OF HLA-B27

几种常见的人类疾病状态，包括强直性脊柱炎 脊柱炎、感染后反应性关节炎和急性前 葡萄膜炎，在遗传上与血清学标记HLA- B27 有说服力的证据表明，一级专业 细胞上组织相容性（MHC）抗原鉴定为HLA-B27 表面直接参与疾病的发病机制。 的 这种参与的机制尚不清楚，但可能与 HLA-B27分子的独特抗原结构。 的 拟议实验的总体目标是扩大 基本上，关于抗原性的现有知识 HLA-B27分子的结构 这将通过四个具体目标来实现： 1)一组突变B27基因将通过以下方法产生： 利多卡因介导的定点诱变;这些突变体 基因将编码B27分子，其氨基酸取代在 预测在同种抗原决定簇中重要的位点 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， 将产生突变的HLAB 27基因或相关的HLA-B7基因 通过显微注射受精卵 3)的单克隆抗体 对HLA-B27的限制性特异性将从转基因产生。 表达B7基因或突变B27基因的小鼠，免疫 用野生型B27转基因小鼠的细胞。 4)这些 抗体，沿着其他单克隆和多克隆抗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 计算机辅助的天然B27上抗原决定因子的图谱 分子，根据最近发表的晶体学地图， HLA-A2. 预计该项目将提供 合理设计的充足资源和信息 成功的实验模型的发病机制B27相关的 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.