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经鼻内免疫 SAG1-弓形虫多聚物纳米球诱导小鼠产生 Th1 型体液免疫应答。

Induction of Th1 type-oriented humoral response through intranasal immunization of mice with SAG1-Toxoplasma gondii polymeric nanospheres.

机构信息

a Department of Parasitology , University of Veterinary and Animal Sciences , Lahore , Pakistan.

b Institute of Biochemistry and Biotechnology , University of Veterinary and Animal Sciences , Lahore , Pakistan.

出版信息

Artif Cells Nanomed Biotechnol. 2018;46(sup2):1025-1034. doi: 10.1080/21691401.2018.1478421. Epub 2018 Jun 6.

Abstract

About one-third of the world population is prone to have infection with T. gondii, which can cause toxoplasmosis in the developing fetus and in people whose immune system is compromised through disease or chemotherapy. Surface antigen-1 (SAG1) is the candidate of vaccine against toxoplasmosis. Recent advances in biotechnology and nano-pharmaceuticals have made possible to formulate nanospheres of recombinant protein, which are suitable for sub-unit vaccine delivery. In current study, the local strain was obtained from cat feces as toxoplasma oocysts. Amplified 957 bp of SAG1 was cloned into pGEM-T and further sub-cloned into pET28-SAG1. BL21 bacteria were induced at different concentrations of isopropyl β-d-1-thiogalactopyranoside for the expression of rSAG1 protein. An immunoblot was developed for the confirmation of recombinant protein expression at 35 kDa that was actually recognized by anti-HIS antibodies and sera were collected from infected mice. PLGA encapsulated nanospheres of recombinant SAG1 were characterized through scanning electron microscopy. Experimental mice were intraperitoneally immunized with rSAG1 protein and intra-nasally immunized with nanosphere. The immune response was evaluated by indirect ELISA. In results intra-nasally administered rSAG1 in nanospheres appeared to elicit elevated responses of specific IgA and IgG2a than in control. Nanospheres of rSAG1 are found to be a bio-compatible candidate for the development of vaccine against T. gondii.

摘要

世界上约有三分之一的人口容易感染刚地弓形虫,这种寄生虫可导致胎儿先天性感染和免疫功能低下人群(如因疾病或化疗而导致免疫功能受损的人群)发生弓形体病。表面抗原-1(SAG1)是弓形虫病疫苗的候选抗原。生物技术和纳米药物学的最新进展使得重组蛋白纳米球的配方成为可能,这种纳米球适合作为亚单位疫苗的载体。在本研究中,本地株是从猫粪便中分离出的刚地弓形虫卵囊。扩增得到的 957bp SAG1 片段被克隆到 pGEM-T 中,并进一步亚克隆到 pET28-SAG1 中。用不同浓度的异丙基 β-d-1-硫代半乳糖苷诱导 BL21 细菌,以表达 rSAG1 蛋白。通过免疫印迹证实了 35kDa 的重组蛋白表达,该蛋白实际上可被抗 HIS 抗体识别,并从感染的小鼠中收集血清。通过扫描电子显微镜对 PLGA 包封的重组 SAG1 纳米球进行了表征。实验小鼠经 rSAG1 蛋白腹腔免疫和纳米球鼻腔免疫,通过间接 ELISA 评估免疫反应。结果表明,鼻腔内给予纳米球包裹的 rSAG1 可引起特异性 IgA 和 IgG2a 的反应升高,优于对照组。纳米球包裹的 rSAG1 被认为是一种具有生物相容性的候选物,可用于开发弓形虫病疫苗。

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