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'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'processes by affecting the CD4+T cells.%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±19.31 vs 23.98±6.56)%,差异具有统计学意义(t=16.66,P<0.0001).IFN-γ在实验组与对照组血清中的表达水平为(39.78±10.77 vs 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±4.03 vs 3.11±0.87)pg/ml,差异具有统计学意义(t=19.75,P<0.0001);IL-1β在实验组与对照组血清中的表达水平为(57.12±12.98 vs 13.45±4.78)pg/ml,差异具有统计学意义(t=19.46,P<0.0001).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.
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