Recombinant vaccinia viruses and gene gun vectors expressing the large subunit of Schistosoma mansoni calpain used in a murine immunization-challenge model
Recombinant vaccinia viruses and gene gun vectors expressing the large subunit of Schistosoma mansoni calpain used in a murine immunization-challenge model
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DOI：
10.1016/s0264-410x(98)00391-0
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发表时间：
1999-03-17
期刊：
影响因子：
5.5
通讯作者：
Dekaban, GA
中科院分区：
文献类型：
--
作者：
Hota-Mitchell, S;Clarke, MW;Dekaban, GA
血吸虫病是一种寄生虫病，每年影响世界热带地区超过 2 亿人。药物治疗和其他现有的控制措施费用昂贵，并且未能消除曼氏血吸虫感染引起的感染、发病和死亡。使用编码关键曼氏血吸虫抗原的重组疫苗对易感个体进行疫苗接种可能是控制血吸虫病的最有效和最便宜的方法。候选疫苗抗原是 p80，曼氏链球菌蛋白酶、钙蛋白酶的大亚基。在我们的疫苗研究中，我们使用了野生型 p80 和突变型 p80 (mut p80)，其中活性位点氨基酸经过基因改造，产生了蛋白水解活性较低的酶。使用p80或mut p80作为疫苗抗原实施了两种疫苗递送方法：重组牛痘病毒(RVV)接种和通过Accell 基因枪(GG)递送系统进行DNA免疫。生成 RVV 的表达 p80 和 mut p80 并测试其体外重组蛋白表达。这些 RW 在小鼠攻击研究中进行了保护能力测试。 RVV-p80 或 RVV-mut p80 皮下或鼻内疫苗接种均不能显着降低接种小鼠的平均蠕虫负荷。在曼氏沙门氏菌攻击感染之前，使用编码p80和mut p80的WRG载体进行GG引发并使用RVV进行加强的GG-RVV组合免疫方案，并且在两次重复研究中没有获得显着的保护。然而，涉及用编码 p80 或 mut p80 的 WRG 载体对小鼠进行 GG 免疫的重复攻击研究表明，对小鼠进行 3 次接种 WRG-full5' mut p80（包含 mut p80 的完整 5' 非翻译区域）可提供 60% 的保护，具有统计学意义 (p < 0.05)。这些初步的体内研究表明，进一步研究基因枪传递的保护作用具有潜力。 WRG-full5' 将 p80 突变到随后受到攻击的小鼠中。此类研究可能为使用表达血吸虫钙激活中性蛋白酶的 WRG DNA 载体对人类进行有效疫苗接种铺平道路。 (C) 1999 Elsevier Science Ltd，保留所有权利。
Schistosomiasis is a parasitic disease affecting over 200 million people every year in tropical regions of the world. Drug treatment and other existing control measures are costly and have failed to eliminate the incidence of infection, morbidity and mortality due to Schistosoma mansoni infection. Vaccination of susceptible individuals using recombinant vaccines encoding key S. mansoni antigens may be the most effective and least expensive means of controlling schistosomiasis. A candidate vaccine antigen is p80, the large subunit of the S. mansoni protease, calpain. In our vaccine studies, we have employed both the wild-type p80 and a mutant p80 (mut p80) in which an active site amino acid was genetically altered to create a less proteolytically-active enzyme. Two vaccine delivery approaches were implemented using p80 or mut p80 as vaccine antigen: recombinant vaccinia virus (RVV) inoculation and DNA immunization via the Accell(R) gene gun (GG) delivery system. RVV's expressing p80 and mut p80 were generated and tested for recombinant protein expression in vitro. These RW's were tested for protective capacity in mouse challenge studies. Neither subcutaneous nor intranasal vaccinations with RVV-p80 or RVV-mut p80 were capable of significantly reducing the mean worm burdens of vaccinated mice. A GG-RVV combination immunization regime using WRG-vectors encoding p80 and mut p80 for GG priming and the RVV's for boosting prior to S. mansoni challenge infection was performed and no significant protection was obtained over two repeated studies. However, duplicate challenge studies involving GG immunization of mice with WRG-vectors encoding p80 or mut p80 revealed that 3 inoculations of mice with WRG-full5' mut p80 (containing the full 5' untranslated region of mut p80) provided 60% protection which was statistically significant (p < 0.05).These preliminary in vivo studies demonstrate the potential for further study of the protection afforded by gene gun-delivered WRG-full5' mut p80 into subsequently-challenged mice. Such studies may pave the way to effective vaccination of humans using WRG DNA vectors expressing a schistosomal calcium-activated neutral protease. (C) 1999 Elsevier Science Ltd, All rights reserved.