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

CD40-CD40配体（CD40L）共刺激已被证明在调节体液和细胞免疫反应中发挥重要作用。在本实验中，我们评估CD40-CD40L系统在SS患者中的作用。采用sCD40L 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例健康对照者的血清。2. 外周血单核细胞（PBMC）染色，双色流式细胞术分析。3. 通过检测CD40L的表达，分别用离子霉素和PMA刺激分离的PBMC。4. 采用抗cd40、CD40L、CD3、CD4、CD8单克隆抗体对SS患者的唇唾液腺进行免疫组织化学染色。5. 提取唇唾液腺总RNA。采用RT-PCR法检测唾液腺CD40和CD40L mRNA的表达。结果：1；I°SS和II°SS患者血清CD40L浓度均高于健康对照组，差异有统计学意义（P<0.01）。2. 健康对照组与SS患者在T细胞和B细胞上CD40L的表达均无显著差异。3. Ionomycin和PMA培养的T细胞在48小时内继续显示CD40L在原代SS中的表达高于正常对照组。4. 免疫组化分析显示，SS患者涎腺浸润性单核细胞中CD40L表达量较少，但表达清晰。5. 在SS患者的唾液腺中检测到CD40和CD40L mRNA的表达，而在正常对照中未检测到。结论：CD40-CD40L系统的激活可能在干燥综合征多克隆B细胞活化的发病机制中起作用。

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.