Wang Lei, Wang Z Q, Cui J
Department of Parasitology, Medical College, Zhengzhou University, PR China.
Vet Parasitol. 2013 May 20;194(2-4):160-3. doi: 10.1016/j.vetpar.2013.01.045. Epub 2013 Feb 5.
The purpose of this study was to determine the protein changes in Trichinella spiralis muscle larvae (ML) after in vitro co-culture with intestinal epithelial cells, and to analyze the changed proteins. Muscle larvae were incubated with HCT-8 cells for 18 h and the ML soluble proteins were analyzed by SDS-PAGE and Western blot. Results showed that seven additional protein bands (123, 77, 58, 36, 30, 28 and 21 kDa) were recognized by sera from infected mice after culture with HCT-8 cells. Three protein bands (97, 51 and 23 kDa) were not recognized by infection sera compared with proteins from ML incubated in RPMI-1640 medium only. Three bands (58, 36 and 21 kDa) were excised from gels, digested with trypsin, and identified by liquid chromatography and tandem mass spectrometry (LC-MS/MS) combined with bioinformatics. In total, 211 kinds of proteins were identified, and numerous enzymes (e.g. metallo- and serine proteases, cysteine proteinases) were found to be highly expressed.
本研究的目的是确定旋毛虫肌幼虫（ML）与肠上皮细胞体外共培养后的蛋白质变化，并分析这些变化的蛋白质。将肌幼虫与HCT-8细胞孵育18小时，通过SDS-PAGE和蛋白质免疫印迹法分析ML可溶性蛋白质。结果显示，与HCT-8细胞共培养后，感染小鼠血清识别出另外七条蛋白带（123、77、58、36、30、28和21 kDa）。与仅在RPMI-1640培养基中孵育的ML蛋白质相比，感染血清未识别出三条蛋白带（97、51和23 kDa）。从凝胶中切下三条带（58、36和21 kDa），用胰蛋白酶消化，通过液相色谱和串联质谱（LC-MS/MS）结合生物信息学进行鉴定。总共鉴定出211种蛋白质，发现许多酶（如金属蛋白酶、丝氨酸蛋白酶、半胱氨酸蛋白酶）高表达。
