Development of recombinant vaccine candidate molecule against Shigella infection
Development of recombinant vaccine candidate molecule against Shigella infection
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DOI：
10.1016/j.vaccine.2016.08.034
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发表时间：
2016-10-17
期刊：
影响因子：
5.5
通讯作者：
Bansal, A.
中科院分区：
文献类型：
--
作者：
Chitradevi, S. T. S.;Kaur, G.;Bansal, A.
志贺菌病是由革兰氏阴性杆菌志贺菌引起的急性细菌性腹泻病。多种志贺氏菌血清型的存在及其对抗生素的耐药性日益增强，迫切需要开发对所有血清型都具有保护性的疫苗。志贺菌IpaB抗原参与易位孔形成，促进细菌侵袭并诱导巨噬细胞凋亡。S.伤寒杆菌GroEL（Hsp 60）是一种免疫优势抗原，可诱导机体产生两种免疫反应，本研究将其作为佐剂进行研究。本研究评估了重组IpaB结构域-GroEL融合蛋白在小鼠中针对致死性志贺氏菌感染的免疫原性和保护功效。用重叠延伸PCR法将IpaB结构域和GroEL基因融合，克隆到pRSETA表达载体中。融合基因在大肠杆菌BL-21细胞中表达，并通过亲和层析纯化所产生的90 KDa融合蛋白。鼻内（i.n.）与IpaB和GroEL免疫组和对照PBS免疫组相比，用融合蛋白免疫小鼠增加了IgG和伊加抗体滴度。融合蛋白免疫组小鼠产生的IgG 1和IgG 2a抗体也高于联合免疫组。淋巴细胞增殖和细胞因子水平（IFN-γ、IL-4和IL-10）的显著增加表明在两个免疫组中诱导了Th 1和Th 2免疫应答。融合蛋白免疫小鼠的保护率为90-95%，而联合免疫组的存活率为80-85%。flexneri，S. boydii和S. sonnei。被动免疫在小鼠中对所有这些志贺氏菌属物种提供60-70%的保护。器官负荷和组织病理学研究还显示，与联合免疫组相比，肺部感染显着减少。由于IpaB是所有志贺菌属物种中保守的优势分子，因此本研究将为开发针对志贺菌血清型的安全、有效和经济的重组疫苗提供理想的平台。(C)2016爱思唯尔有限公司版权所有
Shigellosis is an acute bacillary diarrheal disease caused by the gram negative bacillus Shigella. The existence of multiple Shigella serotypes and their growing resistance to antibiotics stress the urgent need for the development of vaccine that is protective across all serotypes. Shigella's IpaB antigen is involved in translocon pore formation, promotes bacterial invasion and induces apoptosis in macrophages. S. Typhi GroEL (Hsp 60) is the immunodominant antigen inducing both arms of immunity and has been explored as adjuvant in this study. The present study evaluates the immunogenicity and protective efficacy of recombinant IpaB domain-GroEL fusion protein in mice against lethal Shigella infection. The IpaB domain and GroEL genes were fused using overlap extension PCR and cloned in pRSETA expression vector. Fused gene was expressed in Escherichia coli BL-21 cells and the resulting 90 KDa fusion protein was purified by affinity chromatography. Intranasal (i.n.) immunization of mice with fusion protein increased the IgG and IgA antibody titers as compared to the group immunized with IpaB and GroEL and control PBS immunized group. Also IgG1 and IgG2a antibodies induced in fusion protein immunized mice were higher than co-immunized group. Significant increase in lymphocyte proliferation and cytokine levels (IFN-gamma, IL-4 and IL-10), indicates induction of both Thl and Th2 immune responses in both immunized groups. Immunization with fusion protein protected 90-95% of mice whereas 80-85% survivability was observed in co-immunized group against lethal challenge with S. flexneri, S. boydii and S. sonnei. Passive immunization conferred 60-70% protection in mice against all these Shigella species. Organ burden and histopathology studies also revealed significant decrease in lung infection as compared to the co-immunized group. Since IpaB is the conserved dominant molecule in all Shigella species, this study will lead to an ideal platform for the development of safe, efficacious and cost-effective recombinant vaccine against Shigella serotypes. (C) 2016 Elsevier Ltd. All rights reserved.