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

CD40-CD40配体(CD40L)共刺激在调节体液免疫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。方法：1.用sCD40L试剂盒检测33例原发SS(I°SS)、37例继发性SS(II°SS)患者和40例健康对照的血清标本。2.外周血单个核细胞(PBMC)染色及双色流式细胞仪分析。3.分离的PBMC经离子霉素和PMA刺激后，检测CD40L的表达。4.用抗CD40、CD40L、CD3、CD4、CD8单抗对SS患者的唇腺进行免疫组织化学染色。5.从唇腺中提取总RNA。用RT-PCR方法检测唾液腺CD40和CD40L基因的表达。结果：1.I°SS和II°SS患者血清CD40L浓度均显著高于正常对照组(P<0.01)。2.CD40L在T细胞和B细胞上的表达在正常对照组和SS患者之间无显著差异。3.经离子霉素和PMA培养的T细胞在培养48小时后仍可见CD40L在原发SS中的表达高于正常对照。4.SS涎腺的免疫组织化学分析显示，SS患者涎腺中CD40L在浸润性单个核细胞上有少量但清晰的表达。5.在SS患者唾液腺中检测到CD40和CD40L mRNA的表达，而在正常对照组中未检测到CD40和CD40L的表达。结论：CD40-CD40L系统的激活可能参与了干燥综合征多克隆B细胞活化的发病机制。

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.