Yu Hong, Babiuk Lorne A, van Drunen Littel-van den Hurk Sylvia
Vaccine and Infectious Disease Organization, University of Saskatchewan, 120 Veterinary Road, Saskatoon, Sask. S7N 5E3, Canada.
Vaccine. 2007 Feb 26;25(10):1701-11. doi: 10.1016/j.vaccine.2006.11.046. Epub 2006 Dec 5.
In this study, we tested the hypothesis that adoptive transfer of dendritic cells (DCs) transfected ex vivo with mRNA encoding hepatitis C virus (HCV) NS3/4A would initiate potent HCV-specific protective immune responses in vivo. Murine DCs were transfected with NS3/4A mRNA or eGFP mRNA using either electroporation or Transmessenger Transfection Reagent and then used for adoptive transfer. Electroporation resulted in higher transfection efficiency but lower levels of eGFP and NS3/4A expression when compared to transfection with Transmessenger. The murine NS3/4A mRNA-transfected DCs were functional in T cell activation in vitro. Adoptive transfer of NS3/4A mRNA-transfected DCs resulted in migration to regional lymph nodes, strong cellular immune responses and protection from challenge with vaccinia virus expressing NS3/NS4/NS5 in mice. Furthermore, although Transmessenger mediated transfection was less efficient than electroporation in terms of number of transfected cells, the DCs transfected with NS3/4A mRNA and Transmessenger expressed higher levels of protein and induced stronger immune responses and protection than DCs transfected with NS3/4A mRNA by electroporation. Since no study has explored the in vivo efficacy of mRNA-transfected DC-mediated vaccination against viral diseases, including hepatitis C, our study provided a novel vaccination strategy against hepatitis C as well as other pathogens.
在本研究中，我们检验了以下假设：用编码丙型肝炎病毒（HCV）NS3/4A的mRNA体外转染树突状细胞（DC）后进行过继性转移，将在体内引发有效的HCV特异性保护性免疫反应。使用电穿孔或Transmessenger转染试剂将NS3/4A mRNA或eGFP mRNA转染至小鼠DC，然后用于过继性转移。与用Transmessenger转染相比，电穿孔导致更高的转染效率，但eGFP和NS3/4A表达水平较低。小鼠NS3/4A mRNA转染的DC在体外对T细胞激活具有功能性。NS3/4A mRNA转染的DC过继性转移导致其迁移至局部淋巴结，引发强烈的细胞免疫反应，并使小鼠免受表达NS3/NS4/NS5的痘苗病毒攻击。此外，尽管就转染细胞数量而言，Transmessenger介导的转染效率低于电穿孔，但用NS3/4A mRNA和Transmessenger转染的DC比用电穿孔法用NS3/4A mRNA转染的DC表达更高水平的蛋白质，并诱导更强的免疫反应和保护作用。由于尚无研究探讨mRNA转染的DC介导的疫苗接种针对包括丙型肝炎在内的病毒性疾病的体内疗效，我们的研究提供了一种针对丙型肝炎以及其他病原体的新型疫苗接种策略。
