自发性炎性疾病双转基因小鼠的研究

Spontaneous inflammatory diseases in ankylosing spondylitis transgenic mice

目的 通过建立强直性脊柱炎HLA B2 70 4和hβ2 m双转基因动物模型 ,证实HLA B2 70 4和hβ2 m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B2 7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 B2 70 4和hβ2 m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、B2 7单转基因小鼠和HLA B2 7/hβ2 m双转基因小鼠流式细胞计的检测结果分别为 0 6 3%、7 87%、35 87% ,双转基因小鼠的细胞膜表面可见HLA B2 70 4高表达 ,而在B2 7单转基因小鼠表达不明显。 结论 HLA B2 70 4重链可诱发转基因小鼠发生自发性炎性疾病 ,hβ2 m可与B2 7形成稳定的复合体 ,从而稳定和增强HLA B2 70 4在细胞膜表面的表达。

Objective To confirm the role of HLA B2704 and hβ 2m gene in the pathogenesis of spontaneous inflammatory diseases by establishing HLA B2704 and hβ 2m 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 hβ 2m 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 normalmice, B27 single transgenic mice, and HLA B27/ hβ 2m 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 Hβ 2m can form a stable complex with HLA B27 and may stabilize and enhance the expression of HLA B2704 on the cell surface