Identification of novel autoantigens using human IgG in SCID mice implanted with lacrimal and salivary grands from patients with Sjogren's Syndrome

在植入干燥综合征患者泪腺和唾液腺的 SCID 小鼠中使用人 IgG 鉴定新型自身抗原

• 批准号: 16590324
• 负责人: SAKURAI Toshiharu
• 金额: $ 2.3万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590324/
• 关键词: Sjogren's syndrome (SS); salivary and lacrimal glands biopsy tissue; SCID mice; Human IgG antibody; cDNA expression cloning analysis; novel antoantogens

Sjogren's syndrome (SS) is a chronic autoimmune disease characterized by chronic lymphocytic infiltration of the exocrine glands and a worldwide disease with higher prevalence in woman. The salivary and lacrimal glands are the main targets of this lymphocytic infiltration, which as a consequence leads to tissue destruction and decreased function of these glands. But the etiology of SS and mechanisms of lymphocytic infiltration is unknown. The purpose of our project was the identification of specific autoantigens contributing to the disease pathogenesis and progression in SS and useful serological markers for the clinical diagnosis of SS. We demonstrated that the implant of nondisrupted pieces of fresh salivary and lacrimal glands biopsy tissue from patients with SS into the subcutis of SCID mice results in high levels of human IgG (44-3,426μg/ml) produced by gland-infiltrated B and plasma cells in the sera of 20 % mice inoculated with gland biopsy tissue. Using the sera including human IgG from these SCID mice, we demonstrated the cDNA expression cloning analysis with two human salivary gland epithelial cell lines (HSG and HSY) and normal human submandibular grand tissue cDNA libraries in order to identify novel autoantigens as serological marker. The clone encoded SS-A/Ro and SS-B/La, known as a major target of autoantibodies in SS, had the most phage clones isolated by this analysis. To further select the specific autoantigens in SS, we carried out serological screenings using the sera of patients with SS and healthy individuals. Frequencies of IgG autoantibodies against five newerly isolated autoantigens in the sera of patients with SS showed between 30 % and 45 % in significantly higher amounts than in sera from healthy individuals. These results were suggested that our approach using SCID mice may allow the identification of novel specific autoantigens as serological markers for the clinical diagnosis of SS.

干燥综合征（SS）是一种以淋巴细胞浸润外分泌腺为特征的慢性自身免疫性疾病，是一种世界性疾病，女性患病率较高。唾液腺和泪腺是这种淋巴细胞浸润的主要目标，其结果是导致组织破坏和这些腺体的功能降低。但SS的病因和淋巴细胞浸润的机制尚不清楚。本研究的目的是寻找与SS发病机制和进展有关的特异性自身抗原，并寻找临床诊断SS的血清学标志物。我们证明，将来自SS患者的未破裂的新鲜唾液腺和泪腺活检组织植入SCID小鼠皮下组织中，导致20%接种腺体活检组织的小鼠血清中的腺体浸润B和浆细胞产生高水平的人IgG（44- 3，426 μg/ml）。本研究以含人IgG的SCID小鼠血清为材料，利用人涎腺上皮细胞系（HSG和HSY）和正常人下颌下腺组织cDNA文库进行cDNA表达克隆分析，以筛选新的自身抗原作为血清学标记。编码SS-A/Ro和SS-B/La的克隆被称为SS中自身抗体的主要靶标，具有通过该分析分离的最多噬菌体克隆。为了进一步筛选SS的特异性自身抗原，我们用SS患者和健康人的血清进行了血清学筛查。SS患者血清中抗5种新分离的自身抗原的IgG自身抗体的频率在30%至45%之间，显著高于健康个体的血清。这些结果表明，我们的方法使用SCID小鼠可能允许识别新的特异性自身抗原作为临床诊断SS的血清学标志物。

项目成果

Frequent immune response to a melanocyte specific protein KU-MEL-1 in patients with Vogt-Koyanagi-Harada disease

• DOI: 10.1136/bjo.2005.086520
• 发表时间: 2006-02
• 期刊: British Journal of Ophthalmology
• 影响因子: 4.1
• 作者: Soshi otani;Toshiharu Sakurai;Kazuhiko Yamamoto;T. Fujita;Yuriko Matsuzaki;Yasufumi Goto;Yasutaka Ando;Saburosuke Suzuki;Masahiko Usui;Masaru Takeuchi;Yutaka Kawakami