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An Evaluation of accessory molecules in the minor salivary gland from patients with Sjogren's syndrome

干燥综合征患者小唾液腺辅助分子的评估

基本信息

批准号：
12670422
负责人：
OHASHI Hiroyuki
金额：
$ 2.18万
依托单位：
Hamamatsu University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670422/
关键词：
Sjogren's syndrome CD40 CD40L(CD154)Accessory molecules Cytokine Costimulator

项目摘要

CD40-CD40 Ligand (CD40L) costimulation has been shown to play a major role in regulating both humoral and cellular immune responses. In this experiments we evaluated the role of CD40-CD40L system in patients with SS. Methods : 1. Serum samples from 33 primary SS (I°SS) patients, 37 secondary SS (II°SS) patients and 40 healthy controls were measured by sCD40L ELISA kits. 2. Their peripheral blood mononuclear cells (PBMC) were stained and analyzed by two-color flowcytometry. 3. Isolated PBMC were cultured and stimulated by ionomycin and PMA because of detection for CD40L expression. 4. The labial salivary glands from SS patients were immunohistochemically stained by anti-CD40, CD40L, CD3, CD4, CD8 monoclonal antibodies. 5. Total RNA was extracted from labial salivary glands. CD40 and CD40L mRNA expression in salivary glands were detected by RT-PCR method. Results : 1. The concentration of serum CD40L was statistically significant higher (P<0.01) in both I°SS and II°SS patients than in healthy controls. 2. There was no significant difference between healthy controls and SS patients in the expression of CD40L on both T cells and B cells. 3. Cultured T cells by Ionomycin and PMA continued to demonstrate the increased expression of CD40L in primary SS rather than normal controls at 48 hours. 4. Immunohistochemical analyses of SS salivary glands showed that a minor but clear expression of CD40L was present on infiltrating mononuclear cell in the salivary glands from SS patients. 5. The expression of CD40 and CD40L mRNA were detected in the salivary glands from SS patients but not from normal controls. Conclusion : The activation of CD40-CD40L system may play a role of a pathogenesis of polyclonal B cell activation in Sjogren's syndrome.

CD 40-CD 40配体（CD 40 L）共刺激在调节体液和细胞免疫应答中起重要作用。本实验旨在探讨CD_（40）-CD_（40）L系统在SS发病中的作用。方法：1.应用sCD 40 L ELISA试剂盒检测33例原发性SS（I°SS）患者、37例继发性SS（II°SS）患者和40例健康对照者血清sCD 40 L水平。2.采用双色流式细胞仪对患者外周血单个核细胞（PBMC）进行染色和分析。3.分离的PBMC进行培养，并用离子霉素和PMA刺激，因为检测CD 40 L的表达。4.用抗CD_（40）、CD_（40）L、CD_（3）、CD_（4）、CD_（8）单克隆抗体对SS患者唇腺进行免疫组化染色。5.从唇腺中提取总RNA。RT-PCR法检测唾液腺组织中CD 40和CD 40 LmRNA的表达。结果：1. I°SS和II°SS患者血清CD 40 L水平均显著高于健康对照组（P<0.01）。2. SS患者T细胞和B细胞上CD 40 L的表达与健康对照组相比无显著性差异。3.用Ionomycin和PMA培养的T细胞在48小时时继续显示在原发性SS中而不是正常对照中的CD 40 L表达增加。4.免疫组化结果显示，CD 40 L在SS患者涎腺浸润的单个核细胞上有少量但明显的表达。5. CD 40和CD 40 L mRNA在SS患者涎腺中有表达，而在正常对照组中无表达。结论：CD 40-CD 40 L系统的激活可能是干燥综合征多克隆B细胞激活的一个发病机制。