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首页> 外文期刊>Infection and immunity >Development of DNA Vaccines against Hemolytic-Uremic Syndrome in a Murine Model
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Development of DNA Vaccines against Hemolytic-Uremic Syndrome in a Murine Model

机译：在小鼠模型中针对溶血尿毒症综合征的DNA疫苗的开发。

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Shiga toxin type 2 (Stx2) produced by Escherichia coli O:157H7 can cause hemolytic-uremic syndrome in children, a disease for which there is neither a vaccine nor an effective treatment. This toxin consists of an enzymatically active A subunit and a pentameric B subunit responsible for the toxin binding to host cells, and also found to be immunogenic in rabbits. In this study we developed eukaryotic plasmids expressing the B subunit gene of Stx2 (pStx2B) and the B subunit plus the gene coding for the A subunit with an active-site deletion (pStx2ΔA). Transfection of eukaryotic cells with these plasmids produced proteins of the expected molecular weight which reacted with specific monoclonal antibodies. Newborn and adult BALB/c mice immunized with two intramuscular injections of each plasmid, either alone or together with the same vector expressing the granulocyte and monocyte colony-stimulating factor (pGM-CSF), elicited a specific Th1-biased humoral response. The effect of pGM-CSF as an adjuvant plasmid was particularly notable in newborn mice and in pStx2B-vaccinated adult mice. Stx2-neutralizing activity, evaluated in vitro on VERO cell monolayers, correlated with in vivo protection. This is the first report using plasmids to induce a neutralizing humoral immune response against the Stx2.
机译：大肠杆菌O：157H7产生的志贺毒素2型（Stx2）可以引起儿童溶血尿毒症综合征，这种疾病既没有疫苗也没有有效的治疗方法。该毒素由酶促活性A亚基和五聚体B亚基组成，负责毒素与宿主细胞的结合，并且在兔子中具有免疫原性。在这项研究中，我们开发了表达Stx2的B亚基基因（pStx2B）和B亚基以及编码具有活动位点缺失的A亚基的基因（pStx2ΔA）的真核质粒。用这些质粒转染真核细胞产生了预期分子量的蛋白质，该蛋白质与特异性单克隆抗体反应。单独或与表达粒细胞和单核细胞集落刺激因子（pGM-CSF）的相同载体两次肌肉注射每种质粒进行免疫的新生和成年BALB / c小鼠引发了特异性的Th1偏向的体液反应。 pGM-CSF作为佐剂质粒的作用在新生小鼠和接种pStx2B的成年小鼠中尤为明显。在VERO细胞单层体外评估的Stx2中和活性与体内保护作用相关。这是首次使用质粒诱导针对Stx2的中和体液免疫应答的报道。

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