DNA AND CELLULAR STUDIES OF HLA-B27-ASSOCIATED DISEASE

在人类身上的研究。人类白细胞抗原B27与脊柱关节病的关系 疾病将在一组人类白细胞抗原分型的受试者中进行调查， 包括B27+和B27-正常对照，B27+和B27- 脊柱关节病，控制其他风湿性疾病的患者，以及 选定的族。这些人的外周血淋巴细胞 将经历两条基本的实验路线：(1) 将由一组人检测细胞对裂解的敏感性 细胞溶解T淋巴细胞克隆，每个克隆都针对细胞表面的一个表位 人类白细胞抗原B27分子或作为对照抗原决定簇。关联性将 在研究人群中与B-27相关的溶细胞性T细胞 细胞定义的表位与B27相关表位的存在与否 脊椎关节病。这些实验将检验这一假设 T细胞定义的人类白细胞抗原决定簇与 疾病易感性高于血清学定义的人类白细胞抗原-B27。(2)基因组学 从细胞中提取的DNA将被限制消化 内切酶；由此产生的DNA片段将被电泳化 用Southern法分离、印迹和杂交 更多针对人类白细胞抗原-B位点碱基对序列的32P-DNA探针。 将在研究人群中建立被观察到的 限制性内切酶片段长度多态与有无 人类白细胞抗原B27和/或B27相关疾病。这两者的结合 实验方法应该产生很高的概率来确定 脊椎关节病因子是否是B27分子本身 或连锁基因座上的等位基因。B27特异性和/或疾病相关DNA 限制片段将被克隆并进行进一步的 人物刻画。 在老鼠身上的研究。T淋巴细胞在佐剂发病机制中的作用 大鼠关节炎--血清阴性多发性关节炎实验模型 和脊柱炎，将被描述为。致关节炎T细胞系 将在关节炎大鼠的连续培养中建立并评估 关于它们的抗原反应性、体内转运模式和细胞 表面表型。这些动物实验应该能提供对 T淋巴细胞在系统性脊柱炎发病机制中的作用

STUDIES IN HUMANS. The relationship of HLA-B27 to spondyloarthropathic disease will be investigated in a population of HLA-typed subjects, including B27+ and B27- normal controls, B27+ and B27- patients with spondyloarthropathies, control patients with other rheumatic diseases, and selected families. Peripheral blood lymphocytes from these individuals will be subjected to two fundamental lines of experimentation: (1) The cells will be assayed for susceptibility to lysis by a panel of human cytolytic T lymphocyte clones, each specific either for an epitope on the HLA-B27 molecule or for a control antigenic determinant. Correlations will be made within the study population between B-27-associated, cytolytic T cell-defined epitopes and the presence or absence of B27-associated spondyloarthropathic disease. These experiments will test the hypothesis that a T cell-defined HLA determinant is more closely associated with disease susceptibility than is serologically defined HLA-B27. (2) Genomic DNA extracted from the cells will be digested with restriction endonucleases; the resulting DNA fragments will be electrophoretically separated, blotted, and hybridized by the method of Southern with one or more 32P-DNA probes specific for base pair sequences of the HLA-B locus. Correlations will be made within the study population between the observed restriction fragment length polymorphism and the presence or absence of HLA-B27 and/or B27-associated disease. The combination of these two experimental approaches should yield a high probability of determining whether the spondyloarthropathic disease factor is the B27 molecule itself or an allele at a linked locus. B27-specific and/or disease-associated DNA restriction fragments will be cloned and subjected to further characterization. STUDIES IN RATS. The T lymphocytes initiating the pathogenesis of adjuvant arthritis in the rat, an experimental model of seronegative polyarthritis and spondylitis, will be characterized. Lines of arthritogenic T cells will be established in continuous culture from arthritic rats and evaluated for their antigenic reactivity, patterns of trafficking in vivo, and cell surface phenotype. These animal experiments should provide insight into the role of T lymphocytes in the pathogenesis of systemic spondyloarthritis.