摘要目的 对比复发性外阴阴道念珠菌病(RVVC)患者和健康女性树突细胞(DC)表面Dectin-1受体信号传导及功能的差异,分析患者病情反复发作的可能原因.方法 提取1例RVVC患者和1例健康女性的单核细胞,诱导分化为DC.DC与白念珠菌共培养后,流式细胞仪测定DC表面CD83、CD86和CD80表达水平,分析细胞的成熟率;Western印迹法测定DC的Dectin-1、酪氨酸激酶(Syk)和CARD9蛋白的表达;EHSA法测定DC分泌白细胞介素23(IL-23)、肿瘤坏死因子α(TNF-α)和IL-12水平.结果 与健康人DC相比,与白念珠菌共培养24 h后,该RVVC患者DC表面CD83、CD86和CD80的表达活化不明显.与健康人DC相比,与白念珠菌共培养2h后,RVVC患者DC表达的Dectin-1没有显著性差异,但是磷酸化Syk和CARD9活化障碍.与健康人DC相比,与白念珠菌共培养6h后,RVVC患者DC分泌的IL-23、TNF-α和IL-12升高也不明显.抗人Dectin-1抗体对RVVC患者DC的Syk依赖的信号传导通路和上述细胞因子的分泌都没有进一步抑制作用.结论 该RVVC患者DC的Dectin-1受体信号传导通路障碍,导致DC成熟率降低,分泌的IL-23、TNF-α和IL-12降低,使得宿主黏膜抗念珠菌感染的天然免疫功能缺陷.

abstractsObjective To compare the Dectin-1 signal transduction pathway and its function on dendritic cells between a female patient with recurrent vulvovaginal candidiasis (RVVC) and a healthy woman,and to explore the possible mechanism for VVC recurrence in this patient.Methods Venous blood samples were collected from a female patient with RVVC and a healthy woman.Then,monocytes were isolated from the blood samples,and were induced to differentiate into dendritic cells (DCs) in vitro.The obtained DCs were divided into three groups to be cultured alone,cocultured with Candida albicans or the combination of Candida albicans and anti-Dectin-1 antibodies for different durations.Flow cytometry was performed to determine the expression levels of CD83,CD86 and CD80 on DCs to evaluate the maturity of DCs,Western blot analysis to measure the protein expressions of Dectin-1,Syk and CARD9 in DCs,and enzyme-linked immunosorbent assay (ELISA) to determine the levels of interleukin (IL)-23,tumor necrosis factor α (TNF-α) and IL-12 in the culture supernatant of DCs.Results After co-culture with Candida albicans for 24 hours,the expressions of CD83,CD86 and CD80 were significantly inhibited on the patient-derived DCs compared with the controlderived DCs.Western blot analysis showed no significant differences in the expression of Dectin-1 between the controland patient-derived DCs,but a decrease in the expressions of phosphorylated-Syk and CARD9 in the patient-derived DCs compared with the control-derived DCs after 2-hour coculture with Candida albicans.After co-culture with Candida albicans for 6 hours,the levels of IL-23,TNF-α and IL-12 were lower in the culture supernatant of patient-derived DCs than in that of control-derived DCs.Furthermore,the anti-Dectin antibody showed no inhibitory effects on the activation of the Syk-dependent signal transduction pathway in or the secretion of the above cytokines by the patient-derived DCs.Conclusion The Dectin-1 signal transduction pathway was abnormal in DCs from the patient with RVVC,which may decelerate the maturation of DCs,inhibit the secretion of IL-23,TNF-o and IL-12 by them,and finally result in a defect in natural mucosal immunity against Candida infection in the host.