摘要
目的:探讨表达IFN-λ1的重组新城疫病毒rL-hIFN-λ1对巨噬细胞极化的影响。方法:分别用佛波酯、IFN-γ、脂多糖、IL-4和rL-hIFN-λ1刺激人外周血THP-1单核细胞系,构建THP-1-M0、THP-1-M1、THP-1-M2、THP-1-rL-hIFN-λ1型巨噬细胞模型。显微镜下观察M0、M1、M2型巨噬细胞形态特点;实时荧光定量PCR(qRT-PCR)检测各组巨噬细胞M1/M2相关基因表达;免疫荧光检测细胞表面分子CD86、CD163; ELISA检测分泌因子IL-2、IL-13、IL-10及肿瘤坏死因子-α(TNF-α)的表达水平;并采用免疫组织化学法检测不同临床分期肺癌组织中巨噬细胞浸润表型。结果:诱导得到的M0、M1、M2型巨噬细胞形态差异显著。rL-hIFN-λ1诱导的巨噬细胞中,M1型巨噬细胞标志物表达明显升高,M2型巨噬细胞标志物表达显著降低,同时其可促进M1型巨噬细胞因子释放,抑制M2型巨噬细胞因子释放。随着肺癌临床分期进展,M1型巨噬细胞浸润逐渐减少。结论:rL-hIFN-λ1可诱导巨噬细胞向M1型巨噬细胞极化。
Objective: To investigate the effect of recombinant Newcastle disease virus expressing human IFN-λ1( rL-hIFN-λ1) on macrophage polarization. Methods: The THP-1 cells were stimulated with Phorbol 12-myristate 13-acetate( PMA),interferon-γ( IFN-γ)/lipopolysaccharides( LPS),interleukin-4( IL-4) or rL-hIFN-λ1 to construct the THP-1-M0,THP-1-M1,THP-1-M2 and THP-1-rL-hIFN-λ1 macrophage models. The morphological characteristics of M0,M1 and M2 macrophages were observed under the light microscope. The M1/M2 associated gene markers were analyzed by qRT-PCR,the expression of CD86 and CD163 were detected by immunofluorescence analysis,and the releasing cytokines such as IL-2,IL-13,IL-10 and tumor necrosis-α( TNF-α) were examined by ELISA. Furthermore,the M1/M2 infiltration density in different clinical stage of lung cancer tissues were examined by immunohistochemical staining. Results: There were significant differences among the different polarized macrophages. Compared with the THP-1-M0 group,the THP-1-rL-hIFN-λ1 group showed higher expression of M1 biomarkers,and lower M2 biomarkers. The expression of M1 related cytokines was increased,and the M2 related cytokine was decreased. The result of immunohistochemical staining indicated that M1-type macrophages decreased gradually with the development of the clinical stage of lung cancer. Conclusion: rL-hIFN-λ1 could polarize M0 macrophages into M1 macrophages.
作者
张日婷
张垚
章安伟
张烜烽
严玉兰
ZHANG Ri-ting;ZHANG Yao;ZHANG An-wei;ZHANG Xuan-feng;YAN Yu-lan(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 012013;Department of Respiratory,Afiliated Peopled Hospital of Jiangsu University,Zhenjiang Jiangsu 012001,China)
出处
《江苏大学学报（医学版）》
CAS
2018年第6期489-493,共5页
Journal of Jiangsu University：Medicine Edition
基金
江苏省自然科学基金资助项目(BK20151333)
