Tharntada Sirinit, Ponprateep Sirikwan, Somboonwiwat Kunlaya, Liu Haipeng, Söderhäll Irene, Söderhäll Kenneth, Tassanakajon Anchalee
Department of Comparative Physiology, Uppsala University, 75236 Uppsala, Sweden.
Shrimp Molecular Biology and Genomics Laboratory, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
J Gen Virol. 2009 Jun;90(Pt 6):1491-1498. doi: 10.1099/vir.0.009621-0. Epub 2009 Mar 4.
The anti-lipopolysaccharide factor (ALF) from the black tiger shrimp, Penaeus monodon, has been shown previously to exhibit a broad spectrum of activity against various strains of bacteria and fungi. Herein, the recombinant ALFPm3 (rALFPm3) protein was examined for its role in the defence against white spot syndrome virus (WSSV) infection in haematopoietic (Hpt) cell cultures of the freshwater crayfish, Pacifastacus leniusculus, as well as in live P. monodon shrimps. Incubation of Hpt cell cultures with a mixture of WSSV and rALFPm3 resulted in a dose-dependent decrease in VP28 gene expression levels, compared with those incubated with WSSV alone, with an rALFPm3 IC50 value lower than 2.5 microM. However, pre-treatment of Hpt cells with 5 microM rALFPm3 showed no induced protection against subsequent WSSV infection, whereas the synthetic crayfish ALF peptide could protect cells at a higher concentration (10 microM). The in vivo role of ALFPm3 was examined by injection of P. monodon with WSSV pre-treated with rALFPm3 protein. The results clearly showed that rALFPm3 was able to reduce WSSV propagation and prolong the survival of shrimps.
先前研究表明,黑虎虾(斑节对虾)的抗脂多糖因子(ALF)对多种细菌和真菌菌株具有广泛的活性。在此,研究了重组ALFPm3(rALFPm3)蛋白在淡水小龙虾(太平洋螯虾)造血(Hpt)细胞培养物以及活体斑节对虾中抵御白斑综合征病毒(WSSV)感染的作用。与仅用WSSV处理的细胞培养物相比,用WSSV和rALFPm3混合物处理Hpt细胞培养物导致VP28基因表达水平呈剂量依赖性下降,rALFPm3的IC50值低于2.5微摩尔。然而,用5微摩尔rALFPm3预处理Hpt细胞未显示出对随后WSSV感染的诱导保护作用,而合成的小龙虾ALF肽在较高浓度(10微摩尔)下可保护细胞。通过向斑节对虾注射用rALFPm3蛋白预处理的WSSV来研究ALFPm3在体内的作用。结果清楚地表明,rALFPm3能够减少WSSV的传播并延长虾的存活时间。