Marchart J, Dropmann G, Lechleitner S, Schlapp T, Wanner G, Szostak M P, Lubitz W
Institute of Microbiology and Genetics, University of Vienna, Vienna, Austria.
Vaccine. 2003 Sep 8;21(25-26):3988-97. doi: 10.1016/s0264-410x(03)00383-9.
Pasteurella multocida is an important animal pathogen. Bacterial ghosts produced by the expression of phage PhiX174 lysis gene E are empty cells devoid of cytoplasmic and genomic material. Lysis of P. multocida 7A and P. haemolytica A1 carrying Pasteurella-specific lysis vectors (pSR2 and pSON2) occurred 140 min after induction of gene E expression induced by temperature upshift. The E-mediated cell lysis and killing activity was the same in both Pasteurella species and no viable cells could be detected after lysis of P. multocida and P. haemolytica. Pasteurella ghosts were used for immunization of rabbits and mice. Rabbits immunized subcutaneously with either P. multocida- or P. haemolytica-ghosts developed antibodies reacting with the immunizating strain, as well as with other Pasteurella strains. The number of proteins in whole cell protein extracts recognized by the sera constantly increased during the observation period of 51 days. In addition, dose-dependent protection against homologous challenge was observed in mice immunized with P. multocida-ghosts. Animals which received 1.15 x 10(8) ghosts and a challenge dose of up to 60 cfu (LD90), showed 100% protection. According to these results, we suggest ghosts of P. multocida and P. haemolytica as new vaccine candidates.
多杀性巴氏杆菌是一种重要的动物病原体。通过噬菌体PhiX174裂解基因E表达产生的细菌幽灵是空细胞，不含细胞质和基因组物质。携带巴氏杆菌特异性裂解载体（pSR2和pSON2）的多杀性巴氏杆菌7A和溶血巴氏杆菌A1在温度升高诱导基因E表达后140分钟发生裂解。E介导的细胞裂解和杀伤活性在两种巴氏杆菌中相同，多杀性巴氏杆菌和溶血巴氏杆菌裂解后均未检测到活细胞。巴氏杆菌幽灵用于免疫兔和小鼠。用多杀性巴氏杆菌或溶血巴氏杆菌幽灵皮下免疫的兔产生了与免疫菌株以及其他巴氏杆菌菌株反应的抗体。在51天的观察期内，血清识别的全细胞蛋白提取物中的蛋白数量持续增加。此外，在用多杀性巴氏杆菌幽灵免疫的小鼠中观察到对同源攻击的剂量依赖性保护。接受1.15×10(8)个幽灵和高达60 cfu（LD90）攻击剂量的动物显示出100%的保护。根据这些结果，我们建议将多杀性巴氏杆菌和溶血巴氏杆菌的幽灵作为新的疫苗候选物。
