隐球菌脑膜炎患者血清外泌体CD93表达增高及意义

Elevated Expression of CD93 in Serous Exosomes from Patients with Cryptococcal Meningitis and Its Significance

目的检测隐球菌脑膜炎患者血清中外泌体表达CD93的水平,进一步探究其临床意义。方法实验组为2012年11月~2016年12月在上海长海医院和长征医院确诊的38例隐球菌脑膜炎患者的血清,诊断标准为脑脊液染色新生隐球菌阳性或培养阳性;对照组为同期收集的38例健康人血清。血清外泌体的分离采用超高速离心法;外泌体表面抗原的鉴定采用磁珠捕获结合流式细胞仪检测法;血清中细胞因子用ELISA法进行蛋白定量。实验组与对照组检测值的比较采用两独立样本的t检验,两变量之间的相关性以Pearson系数表示。结果流式结果表明,实验组血清外泌体中CD93的表达水平高于对照组为(79.11±19.31vs 23.98±6.56)%,差异具有统计学意义(t=16.66,P<0.000 1)。IFN-γ在实验组与对照组血清中的表达水平为(39.78±10.77vs 58.98±16.99)pg/ml,差异具有统计学意义(t=5.884,P<0.0001);IL-17在实验组与对照组血清中的表达水平为(16.32±4.03vs 3.11±0.87)pg/ml,差异具有统计学意义(t=19.75,P<0.000 1);IL-1β在实验组与对照组血清中的表达水平为(57.12±12.98vs 13.45±4.78)pg/ml,差异具有统计学意义(t=19.46,P<0.000 1)。实验组血清中CD93+exosomes所占的百分比与IFN-γ,IL-17,IL-1β的蛋白表达量呈正相关(r=0.488,0.456,0.532,均P<0.01)。结论隐球菌脑膜炎疾病过程中,CD93在外泌体的表达可能通过影响CD4+T细胞参与了该病病程。

Objective To examine the expression of CD93 in serous exosomes from patients with cryptococcal meningitis,fur-ther explore its clinical significance.Methods The 38 experimental serum samples were from patients who received the diag-nosis with cryptococcal meningitis in Changhai Hospital and Changzheng Hospital in Shanghai from November 2012 to De-cember 2016.The diagnosis standardization was that the cerebrospinal fluid dyeing was positive or the culturing was posi-tive.The 38 controls were collected from the health individuals examined at the same time.The exosomes in serum was sepa-rated by the ultracentrifuge method.Magnetic bead-capture combined with the flow cytometry method was used to identify antigens on the surface of exosomes.The protein level of cytokines in serum was quantified by ELISA method.The compari-son of tested values from experimental and controlled groups was measured by two independent samples＆#39;t test,and the cor-relation between two variates was showed by Pearson coefficient.Results The results of flow cytometry showed that the ex-pression of CD93 on exosomes in the serum samples from experimental group was higher than that in the controlled group, which was（79.11±19.31 vs 23.98±6.56）%,with difference in statistics（t=16.66,P〈0.000 1）.The expression levels of IFN-γin serum samplefrom experimental and control groups were（39.78±10.77 vs 58.98±16.99）pg/ml,with differ-ence in statistics（t=5.884,P〈0.000 1）.The expression levels of IL-17 in serum sample from experimental and control groups were（16.32±4.03 vs 3.11±0.87）pg/ml,with difference in statistics（t=19.75,P〈0.000 1）.The expression lev-els of IL-1β in serum sample from experimental and control groups were（57.12 ± 12.98 vs 13.45 ± 4.78）pg/ml,with difference in statistics（t=19.46,P〈0.000 1）.The percentage of CD93＋exosomes was positively correlative with IFN-γ, IL-17,IL-1β（r=0.488,0.456,0.532,P〈0.01）in serum samples from the experimental group and there was statistical difference.Conclusion In the disease process of cryptococcal meningitis,the expression of CD93 on exosomes might involve in the diseases＆#39;processes by affecting the CD4＋T cells.