摘要
目的研究颗粒酶B(granzymeB,GrB)、颗粒溶素(granulysin,GNLY)及穿孔素(perforin,又称pore-forming pro-tein,PFP)三种免疫效应分子在隐球菌性脑膜炎(cryptococcal meningitis)外周血单个核细胞中的表达及其临床意义。方法首先用密度梯度离心法分离得到25例隐球菌性脑膜炎患者和30例健康人外周血单个核细胞(peripheral blood mononuclear cells,PBMC),其次用免疫印迹(westernblot)的方法检测其中的GrB、GNLY、及PFP的蛋白含量,用实时荧光定量(RFQ)-PCR方法测定其中GrB、GNLY、及PFP的mRNA含量。并分析隐球菌性脑膜炎患者蛋白含量与一些免疫指标的相关性。结果与健康对照组比较,隐球菌性脑膜炎患者PBMC中GNLY、PFP蛋白含量和mRNA均明显降低,而GrB无明显变化。隐球菌性脑膜炎患者中GNLY蛋白表达水平与血清IgG、IFN-γ水平成正相关(r=0.477,P<0.05;r=0.534,P<0.05),与IL-10水平成负相关(r=-0.505,P<0.05)。结论GNLY、PFP可能参与隐球菌性脑膜炎的疾病进程,这为探讨隐球菌性脑膜炎的病情监控和有效治疗提供了新的线索。
Objective To investigate expression of granzyme B（GrB）, granulysin（GNLY） and perforin pefforin（PFP） in PBMC from patients with cryptococcal meningitis and the elinical significances. Methods Peripheral blood mononuclear cells（PBMC） from 25 patients with cryptococcal meningitis and 30 healthy controls were collected and the proteins of GrB, GNLY and PFP were assayed by western blot.Total RNA was extracted from these samples. Real time Fluorescence Quantitative Polymerase Chain Reaction（ FQ- PCR） was performed to assay the level of GrB, GNLY and PFP mRNA. The levels of IL-4 ,IL- 10, TNF-γ and IFN-T in sera were eval- uated by ELISA. The levels of IgG, IgM and IgA in sera were evaluated by routine tests. Results It was showed that the proteins of GNLY and PFP decreased significantly compared with normal controls（relative expression as 0.56 and 0.55, P 〈 0.01 ）. Consistantly, expression of GNLY and PFP mRNA decreased markedly （relative expression as O. 6 and 0.58, P 〈 O. 01）. In addition, expression of GNLY protein was positively related to the levels of scra IgG and IFN-γ（ r = 0.477, P 〈 0.05 ; r = O. 534, P 〈 0.05 ）, while negatively to IL-10（ r = - 0. 505, P 〈 0.05）. Conclusion GNLy and PFP may be involved in the course of cryptococcal meningitis, providing a clue to exploring effective treatment.
出处
《中国实验诊断学》
北大核心
2010年第1期64-68,共5页
Chinese Journal of Laboratory Diagnosis
基金
国家自然科学基金(30671840
30772017
30770997)
上海市科委基金(08QH14001)资助
