α-甘露聚糖肽在诱导人外周血单核细胞来源树突细胞中的作用  

Role of polyactin A in the induction of human peripheral blood monocyte-derived dendritic cells

作　　者：王威[1] 许薇 仝晓宁 王冰 王亚文[1] WANG Wei;XU Wei;TONG Xiaoning;WANG Bing;WANG Yawen(Department of Clinical Laboratory,First Affiliated Hospital,Xi’an Jiaotong University,Xi’an 710061,China;Department of Pathology,School of Basic Medical,Health Science Center,Xi’an Jiaotong University)

机构地区：[1]西安交通大学第一附属医院检验科,西安710061 [2]西安交通大学医学部基础医学院病理学系

出　　处：《山西医科大学学报》2020年第9期927-931,共5页Journal of Shanxi Medical University

基　　金：陕西省重点研发计划项目(2019ZDLSF03-02);西安市科技计划医学研究项目(2019114613YX001SF040);西安交通大学第一附属医院院基金资助项目(2017MS-07)。

摘　　要：目的初步探讨α-甘露聚糖肽(PA)在诱导人外周血单核细胞来源树突细胞(MoDCs)过程中的作用机制,为基于树突细胞的免疫治疗寻找新的经济实用的细胞诱导方案。方法采用磁珠分选法提取外周血浓缩白细胞中的CD14+单核细胞,用于诱导、增殖分化为MoDCs。CD14+单核细胞按细胞因子与PA组合分为:实验组(PA+IL-4+TNF-α)、阳性对照组(GM-CSF+IL-4+TNF-α)和阴性对照组(无细胞因子及PA)。培养过程中,使用倒置显微镜每日观察细胞的生长状态、形态及数量变化情况,记录并采集图像信息。培养至第9天,收集各组细胞,采用流式细胞术进行树突细胞表型鉴定,并对实验数据进行统计学分析。结果经磁珠分选法提取的单核细胞CD14阳性率为(97.40±2.33)%。实验组和阳性对照组在培养过程中具有相似的形态学变化,阴性对照组无细胞增殖及形态学改变。与阴性对照组相比,实验组和阳性对照组CD80^+、CD86^+、HLA-DR+、CD83^+细胞比例明显增高(P<0.05)。与阳性对照组相比,实验组CD80、CD86及HLA-DR的表达差异无统计学意义(P>0.05),CD83的表达减低(P<0.05)。结论PA具有与细胞因子GM-CSF相似的免疫学功能,可参与MoDCs的诱导获得形态典型、特异性表面标志物阳性的树突细胞。Objective To preliminarily investigate the mechanism of polyactin A(PA)inducing human peripheral blood monocyte-derived dendritic cells(MoDCs),and to search a new and economical cell induction scheme for dendritic cell-based immunotherapy.Methods CD14+monocytes were isolated from concentrated leukocytes in human peripheral blood by magnetic bead separation method and used for the induction,the proliferation and differentiation into MoDCs.CD14+monocytes were divided into experimental group(PA+IL-4+TNF-α),positive control group(GM-CSF+IL-4+TNF-α)and negative control group(no cytokines and PA).The growth state and morphological diversification of the cells were observed using an inverted microscope every day.On the 9th day of cell culture,the cells of each group were collected to identify the dendritic cell phenotype by flow cytometry,and the experimental data were statistically analyzed.Results The positive rate of CD14 in monocytes was(97.40±2.33)%.The cells in experimental group and positive control group possessed similar morphologic features of typical mature DCs,while the cells in negative control group showed no obvious morphological changes.Compared with negative group,the proportions of CD80+,CD86+,HLA-DR+and CD83+cells in experimental group and positive control group were increased significantly(P<0.05).Compared with positive control group,there was no significant difference in the expression of CD80,D86 and HLA-DR in experimental group(P>0.05),but the expression of CD83 was decreased(P<0.05).Conclusion PA has the immunological function similar to the cytokine GM-CSF.The strategy of PA involving in MoDCs induction can induce to obtain dendritic cells with typical and specific surface markers positive.

关 键 词：α-甘露聚糖肽 树突细胞 单核细胞 体外诱导 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]