Palaniappan Raghavan U M, McDonough Sean P, Divers Thomas J, Chen Chia-Sui, Pan Ming-Jeng, Matsumoto Mitsuharu, Chang Yung-Fu
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Infect Immun. 2006 Mar;74(3):1745-50. doi: 10.1128/IAI.74.3.1745-1750.2006.
We previously reported the cloning and characterization of leptospiral immunoglobulin-like proteins LigA and LigB of Leptospira interrogans. LigA and LigB are conserved at the amino-terminal region but are variable at the carboxyl-terminal region. Here, we evaluate the potential of recombinant LigA (rLigA) as a vaccine candidate against infection by L. interrogans serovar Pomona in a hamster model. rLigA was truncated into conserved (rLigAcon) and variable (rLigAvar) regions and expressed in Escherichia coli as a fusion protein with glutathione-S-transferase (rLigA). Golden Syrian hamsters were immunized at 3 and 6 weeks of age with rLigA (rLigAcon and rLigAvar) with aluminum hydroxide as an adjuvant. Hamsters given recombinant glutathione-S-transferase (rGST)-adjuvant and phosphate-buffered saline-adjuvant served as nonvaccinated controls. Three weeks after the last vaccination, all animals were challenged intraperitoneally with 10(8) L. interrogans serovar Pomona bacteria (NVSL 1427-35-093002). All hamsters immunized with recombinant LigA survived after challenge and had no significant histopathological changes. In contrast, nonimmunized and rGST-immunized hamsters were subjected to lethal doses, and the hamsters that survived showed severe tubulointerstitial nephritis. All vaccinated animals showed a rise in antibody titers against rLigA. Results from this study indicate that rLigA is a potential vaccine candidate against L. interrogans serovar Pomona infection.
我们之前报道了问号钩端螺旋体的钩端螺旋体免疫球蛋白样蛋白LigA和LigB的克隆及特性。LigA和LigB在氨基末端区域保守,但在羧基末端区域可变。在此,我们在仓鼠模型中评估重组LigA(rLigA)作为抗问号钩端螺旋体波摩那血清型感染疫苗候选物的潜力。rLigA被截短为保守区(rLigAcon)和可变区(rLigAvar),并在大肠杆菌中作为与谷胱甘肽-S-转移酶的融合蛋白表达(rLigA)。金黄叙利亚仓鼠在3周龄和6周龄时用rLigA(rLigAcon和rLigAvar)加氢氧化铝作为佐剂进行免疫。给予重组谷胱甘肽-S-转移酶(rGST)佐剂和磷酸盐缓冲盐水佐剂的仓鼠作为未接种疫苗的对照。最后一次接种后三周,所有动物经腹腔注射10⁸问号钩端螺旋体波摩那血清型细菌(NVSL 1427 - 35 - 093002)进行攻毒。所有用重组LigA免疫的仓鼠在攻毒后存活,且无明显组织病理学变化。相比之下,未免疫和rGST免疫的仓鼠遭受致死剂量,存活的仓鼠出现严重的肾小管间质性肾炎。所有接种疫苗的动物针对rLigA的抗体滴度均升高。本研究结果表明,rLigA是抗问号钩端螺旋体波摩那血清型感染的潜在疫苗候选物。