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融合表达与 DnaK 的钩端螺旋体表面蛋白可诱导小鼠增强免疫应答。

Induction of boosted immune response in mice by leptospiral surface proteins expressed in fusion with DnaK.

机构信息

Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil 1500, 05503-900 São Paulo, SP, Brazil.

Centro de Biotecnologia, Instituto Butantan, Avenida Vital Brazil 1500, 05503-900 São Paulo, SP, Brazil ; Programa Interunidades em Biotecnologia, Instituto de Ciências Biomédicas, USP, Avenida Professor Lineu Prestes 1730, 05508-900 São Paulo, SP, Brazil.

出版信息

Biomed Res Int. 2014;2014:564285. doi: 10.1155/2014/564285. Epub 2014 Jul 6.

Abstract

Leptospirosis is an important global disease of human and veterinary concern. Caused by pathogenic Leptospira, the illness was recently classified as an emerging infectious disease. Currently available veterinarian vaccines do not induce long-term protection against infection and do not provide cross-protective immunity. Several studies have suggested the use of DnaK as an antigen in vaccine formulation, due to an exceptional degree of immunogenicity. We focused on four surface proteins: rLIC10368 (Lsa21), rLIC10494, rLIC12690 (Lp95), and rLIC12730, previously shown to be involved in host-pathogen interactions. Our goal was to evaluate the immunogenicity of the proteins genetically fused with DnaK in animal model. The chosen genes were amplified by PCR methodology and cloned into pAE, an E. coli vector. The recombinant proteins were expressed alone or in fusion with DnaK at the N-terminus. Our results demonstrate that leptospiral proteins fused with DnaK have elicited an enhanced immune response in mice when compared to the effect promoted by the individual proteins. The boosted immune effect was demonstrated by the production of total IgG, lymphocyte proliferation, and significant amounts of IL-10 in supernatant of splenocyte cell cultures. We believe that this approach could be employed in vaccines to enhance presentation of antigens of Leptospira to professional immune cells.

摘要

钩端螺旋体病是一种重要的全球性人兽共患病。由致病性钩端螺旋体引起,该疾病最近被归类为新发传染病。目前可用的兽医疫苗不能诱导长期抗感染保护,也不能提供交叉保护免疫。几项研究表明,由于具有异常高的免疫原性,DnaK 可用作疫苗制剂中的抗原。我们专注于四种表面蛋白:rLIC10368(Lsa21)、rLIC10494、rLIC12690(Lp95)和 rLIC12730,它们先前被证明参与宿主-病原体相互作用。我们的目标是在动物模型中评估与 DnaK 基因融合的蛋白质的免疫原性。选择的基因通过 PCR 方法扩增,并克隆到 pAE 大肠杆菌载体中。重组蛋白单独或与 N 端的 DnaK 融合表达。我们的结果表明,与单独的蛋白质相比,与 DnaK 融合的钩端螺旋体蛋白在小鼠中引起了增强的免疫反应。增强的免疫效应通过脾细胞培养物上清液中总 IgG 的产生、淋巴细胞增殖和大量 IL-10 来证明。我们相信,这种方法可用于疫苗中,以增强对专业免疫细胞的钩端螺旋体抗原的呈递。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5163/4109591/96aae34f6d09/BMRI2014-564285.001.jpg

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