Maes Piet, Keyaerts Els, Bonnet Véronique, Clement Jan, Avsic-Zupanc Tatjana, Robert Alain, Van Ranst Marc
Laboratory of Clinical and Epidemiological Virology, Rega Institute for Medical Research, University of Leuven, Leuven, Belgium.
Intervirology. 2006;49(5):253-60. doi: 10.1159/000093454. Epub 2006 May 22.
We describe the cloning and expression of Dobrava hantavirus (DOBV) nucleocapsid proteins and a truncated form consisting of the first 118 N-terminal amino acids, and the capacity of these E. coli ICONE 200-expressed recombinant proteins (rNp) to induce a protective immune response against DOBV in mice. As an alternative carrier protein, the outer membrane protein A derived from Klebsiella pneumoniae (rP40) has been coupled to different rNp constructs. All recombinant proteins were found to be highly immunogenic after three immunizations of rNp. The immunizations resulted in the induction of a strong Np-specific IgG response with a predominance of IgG1 over IgG2b and IgG2a, suggesting a mixed Th1/Th2 cell involvement. A specific IgG3 response could not be detected. Mice immunized with recombinant DOBV rNp without rP40 showed lower nucleocapsid-specific antibody responses in comparison with the rP40-conjugated constructs, but all mice were found to be protected against DOBV challenge. Our results indicate that the rNp constructs coupled to rP40, represent promising vaccine candidates.
我们描述了多布拉伐汉坦病毒（DOBV）核衣壳蛋白及其由N端前118个氨基酸组成的截短形式的克隆与表达，以及这些在大肠杆菌ICONE 200中表达的重组蛋白（rNp）诱导小鼠产生针对DOBV的保护性免疫反应的能力。作为一种替代载体蛋白，源自肺炎克雷伯菌的外膜蛋白A（rP40）已与不同的rNp构建体偶联。在对rNp进行三次免疫后，发现所有重组蛋白都具有高度免疫原性。这些免疫诱导产生了强烈的Np特异性IgG反应，其中IgG1占主导地位，超过IgG2b和IgG2a，提示有混合的Th1/Th2细胞参与。未检测到特异性IgG3反应。与rP40偶联构建体相比，用不含rP40的重组DOBV rNp免疫的小鼠显示出较低的核衣壳特异性抗体反应，但发现所有小鼠都受到保护，免受DOBV攻击。我们的结果表明，与rP40偶联的rNp构建体是有前景的疫苗候选物。
