感染布氏杆菌后的THP-1细胞的蛋白质组学研究

在布氏杆菌(Brucella)的感染免疫过程中,单核巨噬细胞的应答起着非常关键的作用,而毒力不同的布氏杆菌引起的宿主反应截然不同。用双向电泳技术对THP-1单核细胞受毒力不同的布氏杆菌株侵袭后的全细胞蛋白谱进行差异比较和分析,共发现了38个差异表达的蛋白质点。这些点经过胶内酶切后进行MALDI-TOF质谱鉴定,每个蛋白质点的肽质量指纹图谱都在人类的蛋白质组数据库中用Mascot进行检索后,发现这些差异表达的蛋白主要集中在结构蛋白,信号传导途径和物质代谢等领域,还有一些功能未知的蛋白。这一结果为研究布氏杆菌的感染与致病机制提供了方向,对深入探讨病原菌-宿主的相互作用模式具有参考价值。

Comparative proteomics research on THP-1 cells infected with Brucella

Brucella is a facultative intracellular pathogen that survives and multiplies inside host macrophages to cause brucellosis.The response of macrophage plays an essential role in the initiation of immune process following Brucella challenge.Nowadays,proteome approaches have been widely used in many different systems to investigate host-microbe interactions.The effect of pathogen-specific virulence mechanism can now be dissected using bacterial mutants and comparing different species.Attenuated vaccine strain 104M is defective in multification in host macrophage and is cleared relatively rapidly from tissues of the host, whereas virulent strains Brucella abortus 544A can produce chronic infection and cause brucellosis.In order to understand the underlying mechanisms of virulent Brucella intracellular survival,and detect the different-expressed proteins of THP-1 cells after infection with attenuated and virulent strains of Brucella abortus,a comparative proteomics research was conducted.Whole cellular protein profiling of THP-1 cells was presented by two dimensional(2D) electrophoresis and Coomassie Blue staining.After in-gel protein digestion,the different-expressed spots were detected by matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS).All the peptide mass fingerprints(PMFs) were searched by the program Mascot developed by Matrix Science Ltd.For identifying proteins,database of hemo sapiens was used.A total of 38 proteins with changed expression level were found.These proteins can be grouped into two familes:1)the expression level increased after infection with 544A;2) the expression level increased after infection with 104M.Out of the 38 proteins,10 were mainly in the field of signal transduction,6 were cytoskeletal proteins,8 were substance metabolism related proteins and 3 were cell stress and defense associated proteins.Functions of the remaining proteins were unknown.These results provide insight into the changed global protein patterns of THP-1 cells after infection as well as a comprehensive foundation to further study of host-bacterial interaction.