自发性炎性疾病双转基因小鼠的研究

目的 通过建立强直性脊柱炎HLA－B2704和hβ2m双转基因动物模型,证实HLA－B2704和hβ2m基因在双转基因小鼠自发性炎性疾病发病过程中的作用,为研究B27相关性疾病的病因学、预防和治疗提供有力的工具。方法 将HLA－B2704和hβ2m转基因阳性小鼠交配后出生的子代,应用PCR、斑点杂交、Southern杂交、RT－PCR、流式细胞术和免疫组织化学等方法进行筛选鉴定和表达检测。同时隔日观察小鼠的发病情况,发病鼠行HE染色观察病理变化。结果 有8只高拷贝双转基因小鼠2周左右出现皮肤病变,关节炎和趾甲变化等自发性炎性疾病表现,正常小鼠、B27单转基因小鼠和HLA－B27／hβ2m双转基因小鼠流式细胞计的检测结果分别为0．63％、7．87％、35．87％,双转基因小鼠的细胞膜表面可见HLA－B2704高表达,而在B27单转基因小鼠表达不明显。结论 HLA－B2704重链可诱发转基因小鼠发生自发性炎性疾病,hβ2m可与B27形成稳定的复合体,从而稳定和增强HLA－B2704在细胞膜表面的表达。

Spontaneous inflammatory diseases in ankylosing spondylitis transgenic mice

OBJECTIVE: To confirm the role of HLA-B2704 and hbeta(2)m gene in the pathogenesis of spontaneous inflammatory diseases by establishing HLA-B2704 and hbeta(2)m double transgenic mice model of ankylosing spondylitis. It will provide a powerful animal model for exploring the etiology, prevention and treatment of B27-relevant diseases. METHODS: The screening, identification and expression of HLA-B2704 and hbeta(2)m gene were determined by PCR, dot blot, Southern blot hybridization, RT-PCR, flow cytometry and immunohistochemistry. HE staining was performed for the diseased mice. RESULTS: Eight double transgenic mice bearing high copy developed spontaneous dermatosis, arthritis and nail changes in the rear paw. The results of flow cytometry in normal mice, B27 single transgenic mice, and HLA-B27/hbeta(2)m double transgenic mice were 0.63%, 7.87% and 35.87% respectively. HLA-B2704 antigen was high expressed on the cell surface, but not evident on those of B27 single transgenic mice. CONCLUSIONS: HLA-B2704 heavy chain can induce spontaneous inflammatory diseases in the transgenic mice. Hbeta(2)m can form a stable complex with HLA-B27 and may stabilize and enhance the expression of HLA-B2704 on the cell surface.