McEwen J, Levi R, Horwitz R J, Arnon R
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Vaccine. 1992;10(6):405-11. doi: 10.1016/0264-410x(92)90071-q.
The influenza virus haemagglutinin epitope 91-108, which is a conserved amino acid sequence in all type A H3 strains, was expressed in Salmonella flagellin, to evaluate its potential as a vaccine. For that purpose, a synthetic oligonucleotide comprising 54 bases coding for the corresponding sequence was inserted into the plasmid pLS408 and transformed into Escherichia coli JM101. Colonies containing the recombinant plasmid were used to transform Salmonella typhimurium LB5000 and were then transduced to a flagellin negative 'live vaccine' aroA mutant of Salmonella dublin. Rabbits immunized either with the live recombinant S. dublin or with the flagellin isolated from it, showed significant levels of IgG response against the synthetic peptide 91-108 as well as against the intact A/Texas/77 influenza virus. Mice immunized with the same preparations developed influenza-specific IgG antibodies in the blood and secreted IgA antibodies in their lungs. Furthermore, these mice showed about 50% protection against challenge infection with the virus. The most successful results were achieved by intranasal immunization with the isolated recombinant flagellin, when employed without the aid of adjuvant.
流感病毒血凝素表位91 - 108是所有甲型H3毒株中的一个保守氨基酸序列，它在沙门氏菌鞭毛蛋白中表达，以评估其作为疫苗的潜力。为此，将一个包含54个碱基、编码相应序列的合成寡核苷酸插入质粒pLS408，并转化到大肠杆菌JM101中。含有重组质粒的菌落用于转化鼠伤寒沙门氏菌LB5000，然后转导至都柏林沙门氏菌鞭毛蛋白阴性的“活疫苗”aroA突变体。用活的重组都柏林沙门氏菌或从其中分离出的鞭毛蛋白免疫的兔子，对合成肽91 - 108以及完整的A/德克萨斯/77流感病毒显示出显著水平的IgG反应。用相同制剂免疫的小鼠在血液中产生了流感特异性IgG抗体，并在肺部分泌了IgA抗体。此外，这些小鼠对病毒攻击感染表现出约50%的保护作用。当不使用佐剂时，通过鼻内免疫分离出的重组鞭毛蛋白取得了最成功的结果。
