Humphryes P C, Weeks M E, AbuOun M, Thomson G, Núñez A, Coldham N G
Animal Health and Veterinary Laboratories Agency, New Haw, Addlestone, Surrey, United Kingdom.
Clin Vaccine Immunol. 2014 Apr;21(4):546-51. doi: 10.1128/CVI.00719-13. Epub 2014 Feb 12.
The Leptospira interrogans vaccines currently available are serovar specific and require regular booster immunizations to maintain protection of the host. In addition, a hamster challenge batch potency test is necessary to evaluate these vaccines prior to market release, requiring the use of a large number of animals, which is ethically and financially undesirable. Our previous work showed that the N terminus of the outer membrane protein LipL32 was altered in Leptospira interrogans serovar Canicola vaccines that fail the hamster challenge test, suggesting that it may be involved in the protective immune response. The aim of this study was to determine if vaccination with LipL32 protein alone could provide a protective response against challenge with L. interrogans serovar Canicola to hamsters. Recombinant LipL32, purified from an Escherichia coli expression system, was assessed for protective immunity in five groups of hamsters (n = 5) following a challenge with the virulent L. interrogans serovar Canicola strain Kito as a challenge strain. However, no significant survival against the L. interrogans serovar Canicola challenge was observed compared to that of unvaccinated negative controls. Subsequent histological analysis revealed reduced amounts of L. interrogans in the kidneys from the hamsters vaccinated with recombinant LipL32 protein prior to challenge; however, no significant survival against the L. interrogans serovar Canicola challenge was observed compared to that of unvaccinated negative controls. This finding corresponded to a noticeably reduced severity of renal lesions. This study provides evidence that LipL32 is involved in the protective response against L. interrogans serovar Canicola in hamsters and is the first reported link to LipL32-induced protection against kidney invasion.
目前可用的问号钩端螺旋体疫苗具有血清型特异性,需要定期加强免疫以维持对宿主的保护。此外,在疫苗上市前,需要进行仓鼠攻毒批效价试验来评估这些疫苗,这需要使用大量动物,从伦理和经济角度来看都不理想。我们之前的研究表明,在仓鼠攻毒试验中失败的问号钩端螺旋体犬型血清型疫苗中,外膜蛋白LipL32的N端发生了改变,这表明它可能参与了保护性免疫反应。本研究的目的是确定单独用LipL32蛋白接种疫苗是否能为仓鼠提供针对问号钩端螺旋体犬型血清型攻毒的保护性反应。从大肠杆菌表达系统中纯化的重组LipL32,在五组仓鼠(n = 5)中用强毒问号钩端螺旋体犬型血清型菌株Kito作为攻毒菌株进行攻毒后,评估其保护性免疫。然而,与未接种疫苗的阴性对照组相比,未观察到对问号钩端螺旋体犬型血清型攻毒有显著的存活差异。随后的组织学分析显示,在攻毒前用重组LipL32蛋白接种的仓鼠肾脏中问号钩端螺旋体的数量减少;然而,与未接种疫苗的阴性对照组相比,未观察到对问号钩端螺旋体犬型血清型攻毒有显著的存活差异。这一发现与肾脏病变严重程度明显降低相对应。本研究提供了证据,表明LipL32参与了仓鼠对问号钩端螺旋体犬型血清型的保护性反应,并且是首次报道的LipL32诱导的针对肾脏侵袭的保护作用的联系。