Feng Hao, Liu Lefeng, Chi Jiaqi, Wang Ning, Li Runting, Tong Chunyu, Ma Jinzhu, Zhu Zhanbo, Cui Yudong
School of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Aug;25(8):1180-6.
In order to characterize the immunogenicity and immunoprotection of the Staphylococcus aureus (S. aureus) surface protein Clumping factor A (ClfA), we amplified clfa genes from S. aureus Newman strain, Wood46 strain and HLJ23-1. The clfa gene from Newman strain was subsequently inserted into pQE-30 vector and the recombinant plasmid was transformed into Escherichia coli strain M15 (pREP4). The recombinant ClfA protein was expressed and purified. Then, we immunized mice with the purified recombinant protein. The antibody level and the concentration of cytokines were measured by enzyme-linked immunosorbent assay. Finally, immunized mice were challenged with S. aureus Newman, Wood46 and HLJ23-1. These results suggested that clfa gene sequences were highly conserved, and the recombinant ClfA was expressed correctly with good antigenicity. The antibody titer and the concentration of cytokines in the immunized groups increased significantly (P < 0.05) compared with control, and the mice in the immunized groups were protected against the challenge strains to some extent. These results showed that the ClfA had high immunogenicity and immunoprotective potential.
为了表征金黄色葡萄球菌（S. aureus）表面蛋白凝聚因子A（ClfA）的免疫原性和免疫保护作用，我们从金黄色葡萄球菌纽曼菌株、伍德46菌株和HLJ23 - 1中扩增了clfa基因。随后将纽曼菌株的clfa基因插入pQE - 30载体，并将重组质粒转化到大肠杆菌菌株M15（pREP4）中。表达并纯化了重组ClfA蛋白。然后，我们用纯化的重组蛋白免疫小鼠。通过酶联免疫吸附测定法测量抗体水平和细胞因子浓度。最后，用金黄色葡萄球菌纽曼菌株、伍德46菌株和HLJ23 - 1对免疫小鼠进行攻毒。这些结果表明clfa基因序列高度保守，重组ClfA正确表达且具有良好的抗原性。与对照组相比，免疫组的抗体效价和细胞因子浓度显著升高（P < 0.05），并且免疫组的小鼠在一定程度上对攻毒菌株具有抵抗力。这些结果表明ClfA具有高免疫原性和免疫保护潜力。
