Vemulapalli Ramesh, Contreras Andrea, Sanakkayala Neelima, Sriranganathan Nammalwar, Boyle Stephen M, Schurig Gerhardt G
Department of Veterinary Pathobiology, School of Veterinary Medicine, Purdue University, 725 Harrison Street, West Lafayette, IN 47907, USA.
Vet Microbiol. 2004 Sep 8;102(3-4):237-45. doi: 10.1016/j.vetmic.2004.07.001.
Brucella abortus strain RB51 is an attenuated rough strain, currently being used as the official live vaccine for bovine brucellosis in the USA and several other countries. In strain RB51, the wboA gene, encoding a glycosyltransferase required for the O-side chain synthesis, is disrupted by an IS711 element. Recently, we have demonstrated that strain RB51WboA, RB51 complemented with a functional wboA gene, remains rough but expresses low quantities of O-side chain in the cytoplasm. Mice vaccinated with strain RB51WboA develop greatly enhanced resistance against challenge with B. abortus virulent strain 2308. We have also demonstrated that overexpression of Cu/Zn superoxide dismutase (SOD) in strain RB51 (RB51SOD) significantly increases its vaccine efficacy against strain 2308 challenge. In this study, we constructed a new recombinant strain, RB51SOD/WboA, that over expresses SOD with simultaneous expression of O-side chain in the cytoplasm. We tested the vaccine potential of strains RB51SOD, RB51WboA, RB51SOD/WboA against challenge with virulent Brucella melitensis 16M and B. abortus 2308 in mice. In comparison with strain RB51, strain RB51SOD induced better protection against strain 2308, but not strain 16M, challenge. Similar to strain RB51WboA, vaccination with strain RB51SOD/WboA resulted in complete protection of the mice from infection with strain 2308. When challenged with strain 16M, mice vaccinated with either strain RB51WboA or strain RB51SOD/WboA were significantly better protected than those vaccinated with strain RB51 or RB51SOD. These results suggest that strains RB51WboA and RB51SOD/WboA are good vaccine candidates for inducing enhanced protection against B. melitensis infection.
布鲁氏菌流产株RB51是一种减毒粗糙型菌株,目前在美国和其他几个国家用作牛布鲁氏菌病的官方活疫苗。在RB51菌株中,编码O侧链合成所需糖基转移酶的wboA基因被一个IS711元件破坏。最近,我们已经证明,用功能性wboA基因互补的RB51WboA菌株仍然是粗糙型,但在细胞质中表达少量的O侧链。用RB51WboA菌株接种的小鼠对流产布鲁氏菌强毒株2308的攻击产生了大大增强的抵抗力。我们还证明,RB51菌株(RB51SOD)中铜/锌超氧化物歧化酶(SOD)的过表达显著提高了其对2308菌株攻击的疫苗效力。在本研究中,我们构建了一种新的重组菌株RB51SOD/WboA,其在细胞质中同时表达O侧链的情况下过表达SOD。我们测试了RB_51SOD、RB51WboA、RB51SOD/WboA菌株对小鼠感染强毒布鲁氏菌16M和流产布鲁氏菌2308攻击的疫苗潜力。与RB51菌株相比,RB51SOD菌株对2308菌株的攻击诱导了更好的保护作用,但对16M菌株的攻击则不然。与RB51WboA菌株类似,用RB51SOD/WboA菌株接种可使小鼠完全免受2308菌株的感染。当用16M菌株攻击时,用RB51WboA菌株或RB51SOD/WboA菌株接种的小鼠比用RB51或RB51SOD菌株接种的小鼠受到的保护明显更好。这些结果表明,RB51WboA和RB51SOD/WboA菌株是诱导增强抗布鲁氏菌16M感染保护作用的良好疫苗候选株。